Molecular Biology Reports (03014851)50(1)pp. 517-530
Background: Myocardial infarction-associated transcript (MIAT) is a long non-coding RNA (lncRNA) with altered expression in different diseases and malignancies. In this study, the potential expression and function of lncRNA MIAT in intuition and progression of brain cancer was investigated. Methods and Results: At first, TCGA data analysis demonstrated that lncRNA MIAT is significantly upregulated in various malignancies, especially its expression is dramatically elevated in brain tumors. In line with the data, we further evaluated the expression of MIAT in a series of brain tumor tissue, and our results revealed that the expression of MIAT was noticeably overexpressed in glioblastoma (p = < 0.0001). We further found that the expression of MIAT was markedly upregulated in low-grade brain tumors rather than high-grade ones. To further investigate the biological function of MIAT in brain cancer cells, its expression was suppressed by si-RNA-mediated knocking down. Inhibition of MIAT resulted in reduced proliferation of brain tumor cells followed by cell cycle arrest at the G1 phase, and significant induction of apoptosis, and senescence, but limited the migration ability and epithelial-mesenchymal-transition (EMT). Moreover, knocking-down of MIAT reduced the expression of stemness factors, followed by upregulation of their downstream miRNAs (micro RNAs), let-7a-5p, and miR-29b-3p. Conclusions: Altogether, our data demonstrated that lncRNA MIAT could control proliferation, migration, and metastasis of brain cancer cells via regulating the Nanog/ Sox2 / let-7a-5p / miR-29b-3p axis. This data could introduce lncRNA MIAT as a novel oncogene in brain cancer pathogenesis. © 2022, The Author(s), under exclusive licence to Springer Nature B.V.
Garavaglia, B.,
Vallian borujeni, S.,
Romito, L.M.,
Straccia, G.,
Capecci, M.,
Invernizzi, F.,
Andrenelli, E.,
Kazemi, A.,
Boesch, S.,
Kopajtich, R. Parkinsonism and Related Disorders (18735126)97pp. 52-56
Introduction: The genetic basis of autosomal-recessive dystonia remains poorly understood. Our objective was to report identification of additional individuals with variants in AOPEP, a recently described gene for recessively inherited dystonic disorders (OMIM:619565). Methods: Ongoing analysis on a high-throughput genetic platform and international case-recruitment efforts were undertaken. Results: Novel biallelic, likely pathogenic loss-of-function alleles were identified in two pedigrees of different ethnic background. Two members of a consanguineous Iranian family shared a homozygous c.1917-1G>A essential splice-site variant and featured presentations of adolescence-onset generalized dystonia. An individual of Chinese descent, homozygous for the nonsense variant c.1909G>T (p.Glu637*), displayed childhood-onset generalized dystonia combined with later-manifesting parkinsonism. One additional Iranian patient with adolescence-onset generalized dystonia carried an ultrarare, likely protein-damaging homozygous missense variant (c.1201C>T [p.Arg401Trp]). Conclusions: These findings support the implication of AOPEP in recessive forms of generalized dystonia and dystonia-parkinsonism. Biallelic AOPEP variants represent a worldwide cause of dystonic movement-disorder phenotypes and should be considered in dystonia molecular testing approaches. © 2022 Elsevier Ltd
BioFactors (09516433)48(1)pp. 164-180
Long noncoding RNAs (lncRNAs) appear as vital regulators and biomarkers in many human cancers. LOC100507144 is a validated lncRNA located in the neighborhood of CD44 in a head-to-head configuration, and its expression and function in cancer cells are still unknown. This research aimed to find out more about the expression and function of this lncRNA in colorectal cancer (CRC). Our expression data represented that the expression of LOC100507144 transcript was substantially higher in tumors with advanced stages, lymph node metastasis, and vascular invasion. Loss-of-function examinations demonstrated that LOC100507144 contributed to CRC cell proliferation by restricting apoptosis, cellular senescence, and promoting cell cycle. Gain-of-function experiments also confirmed these results. Our data illustrated that LOC100507144 enhanced the migration and the epithelial to mesenchymal transition (EMT) of CRC cells, accompanied by the generation of cells with stemness characteristics. Our findings revealed that the knocking-down of LOC100507144 inhibited the expression of crucial stemness factors, including CD44, Nanog, and Sox2, and accordingly resulted in suppressing their targets, miR-302 and miR-21. Overall, the current study's findings for the first time reveal that LOC100507144 could enhance CRC progression and metastasis through regulation of the CD44/Nanog/Sox2/miR-302/miR-21 axis. © 2021 International Union of Biochemistry and Molecular Biology.
Scientific Reports (20452322)11(1)
The pathways and robust deregulated gene signatures involved in AML chemo-resistance are not fully understood. Multiple subgroups of AMLs which are under treatment of various regimens seem to have similar regulatory gene(s) or pathway(s) related to their chemo-resistance phenotype. In this study using gene set enrichment approach, deregulated genes and pathways associated with relapse after chemotherapy were investigated in AML samples. Five AML libraries compiled from GEO and ArrayExpress repositories were used to identify significantly differentially expressed genes between chemo-resistance and chemo-sensitive groups. Functional and pathway enrichment analysis of differentially expressed genes was performed to assess molecular mechanisms related to AML chemotherapeutic resistance. A total of 34 genes selected to be differentially expressed in the chemo-resistance compared to the chemo-sensitive group. Among the genes selected, c-Jun, AKT3, ARAP3, GABBR1, PELI2 and SORT1 are involved in neurotrophin, estrogen, cAMP and Toll-like receptor signaling pathways. All these pathways are located upstream and regulate JNK signaling pathway which functions as a key regulator of cellular apoptosis. Our expression data are in favor of suppression of JNK pathway, which could induce pro-apoptotic gene expression as well as down regulation of survival factors, introducing this pathway as a key regulator of drug-resistance development in AML. © 2021, The Author(s).
Archives of Biological Sciences (03544664)73(4)pp. 457-463
The presence of single nucleotide variations in the coding region of micro-RNA (miRNA)-encoding genes plays a significant role in the expression and function of these molecules in oncogenesis and cancer. The association of rs2682818 in miR-618 with increased risk of breast cancer was investigated in the Iranian population. rs2682818/miR-618 was genotyped using amplification-refractory mutation system PCR (ARMS-PCR) in 200 healthy individuals and patients with breast cancer. The data revealed the presence of Hardy-Weinberg equilibrium (HWE) for this marker. The frequency of alleles C and A was 70% and 30%, respectively, in healthy individuals; the frequency of alleles C and A was 44% and 56%, respectively, in patients with breast cancer. Analysis of odd ratios showed that the rs2682818/miR-618 polymorphism is associated with increased probability of breast cancer and is statistically significant (OR=2.97, P=0.0003). The data suggest that rs2682818/miR-618 could be considered a novel marker of increased risk of breast cancer © 2021. by the Serbian Biological Society
Tay-Sachs Disease (TSD) is a genetic disorder resulting from mutations in the HEXA gene. Linkage analysis of polymorphic markers such as single nucleotide polymorphism (SNP) has been used in heterozygous carrier detection and prenatal diagnosis of the disease in families with an affected individual. A large number of SNP markers have been introduced for the HEXA gene in the databases. In the present study, the genotype and informative degree of rs2288258 and rs4777507genetic markers of HEXA gene region were investigated. This study has been conducted using Tetra-primer ARMS PCR and usual ARMS PCR in 148 healthy unrelated control individuals and 10 families in the Iranian population. The allele frequency, degree of heterozygosity and Hardy-Weinberg equilibrium (HWE) was estimated using the GENEPOP program. Also, the estimation of haplotype frequency and linkage disequilibrium for the unrelated individuals was performed using the PowerMmarker software. Based on the obtained results, the observed degree of heterozygosity for rs2288258 and rs4777507 were 18.24 and 27.70, respectively. The allele frequency of rs2288258 was 90.88% for allele A and 9.12% for allele G. Also, the allele frequency of rs4777507 was 84.8% and 15.2% for A and G allele respectively. The results showed two markers, rs2288258-rs4777507, were in linkage disequilibrium together. Consequently, A/A, A/G and G/G informative haplotypes in population level and A/A, A/G and G/A informative haplotypes in family level were identified by using a combination of these two markers. These haplotypes can serve as appropriate tools for linkage analysis and identification of heterozygous carriers and prenatal diagnosis of TSD in the Iranian families with affected children. © 2020
Journal of Genetics (09737731)99(1)
Nephrotic syndrome (NS) is considered as a primary disease of the kidney that represents a heterogeneous group of glomerular disorders occurring mainly in children. It is generally divided into steroid-sensitive and steroid-resistant forms, depending upon the patient’s response to steroid therapy. Among the genes involved, the NPHS2 gene has been reported as the causative gene in steroid resistant form of nephrotic syndrome. In the present study, heterozygosity rate, allelic frequency and linkage of rs2274625 and rs3829795 markers were investigated in the NPHS2 gene region. To determine the SNP alleles, tetra-primer ARMS PCR was used. After genotyping rs2274625 and rs3829795 polymorphic markers in 120 unrelated individuals and nine trios families, the data were analysed using various computer programs such as UCSC Genome Browser, dbSNP and SNPper. Based on the statistical analysis of the results, for rs2274625 marker, allele frequency for C and T alleles was 97% and 3%, respectively. For rs3829795 marker allele frequency for G and A alleles was 55% and 45%, respectively. The values of heterozygosity index for the examined markers were 5% for rs2274625 and 45/8% for rs3829795. Consequently, two informative haplotypes, CG/CA, were identified in the NPHS2 gene region through combination of these two markers. These haplotypes can serve as appropriate tools for the identification of heterozygous carriers and linkage analysis of nephrotic syndrome disease in the Iranian families with an affected child. © 2020, Indian Academy of Sciences.
Cell Journal (Yakhteh) (22285806)22pp. 110-116
Objective: Thirteen million cancer deaths and 21.7 million new cancer cases are expected in the world by 2030. Breast cancer is considered as the main cause of cancer mortality in women aged 20-59 years. microRNAs (miRNAs) regulate gene expression at the post-transcriptional level and they are highly expressed in malignancies, including breast cancer. The role of miRNAs in the pathogenesis of breast cancer is not fully understood. In the present study, for the first time, the impact of hsa-miR-423 rs6505162 on breast cancer risk was investigated in the central province of Iran, Isfahan. Materials and Methods: This case-control study was conducted on 153 clinicopathological proven breast cancer patients and 153 sex-matched healthy women with no history of any cancer type and relative patients. The patients and controls were genotyped and association of their clinical characteristics with hsa-miR-423 rs6505162 genotype was analyzed. Results: The findings indicated that CC genotype of hsa-miR-423 rs6505162 was associated with the increased risk of breast cancer [odds ratio (OR)=2.37, 95% confidence interval (CI)=1.29-4.35 and P=0.0023, CC vs. AA]. Conclusion: The data suggested that hsa-miR-423 rs6505162 could be considered as a novel risk factor in breast cancer pathogenesis in Isfahan province of Iran. © 2020 Royan Institute (ACECR). All rights reserved.
Niemann-Pick disease (NPD) refers to a group of lysosomal storage diseases that cause abnormal metabolism of lipids. Direct sequencing and deletion analysis is used to detect mutations and sequence variations in the SMPD1 gene. However, due to technical difficulties, linkage analysis of polymorphic markers such as single nucleotide polymorphism (SNP) has been used in heterozygous carrier detection and prenatal diagnosis of the disease in families with an affected individual. The SNP markers usually show a population-based haplotype frequency and heterozygosity. A large number of SNP markers have been introduced for the SMPD1 gene. In the present study, rs1542705, rs67992843and rs1050239 genetic markers located in the SMPD1 gene region were genotyped using ARMS PCR technique in 188 unrelated control individuals and 11 family trios in the Iranian population. The allele frequency, degree of heterozygosity and Hardy-Weinberg equilibrium (HWE) were estimated using the GENEPOP program. The estimation of haplotype frequency and linkage disequilibrium for the unrelated individuals was estimated using the PowerMarker software. The observed degree of heterozygosity for rs1542705, rs6799843and rs1050239 were 39.36, 46.68 and 28.19, respectively. The result showed all markers were in linkage disequilibrium. Consequently, T-G-G, T-C-G, C-G-G, C-C-G, T-C-A, T-G-A, C-G-A informative haplotypes in population level were identified by using a combination of these three markers. Therefore, 1,542,705–6,799,843-1,050,239 represents a novel informative haplotype at the SMPD1 locus in the Iranian population, which could be suggested as appropriate markers for linkage analysis and identification of heterozygous carriers as well as prenatal diagnosis of NPD in the Iranian families with affected children. © 2020
European Journal of Medical Genetics (18780849)62(9)
Phenylketonuria (PKU) is a metabolic disorder caused by mutations in the phenylalanine hydroxylase (PAH) gene. After thalassemia, PKU is considered as the most common autosomal recessive diseases in the Iranian population. Therefore, an efficient diagnostic strategy is required to identify disease-causing mutations in this population. Following our first report in 2003, here we presented a comprehensive study on the mutation spectrum of the PAH gene in the Iranian population. This study was performed on 280 unrelated chromosomes from 140 Iranian patients with classic PKU. All 13 exons as well as exon-intron boundaries of the PAH gene were analyzed by direct DNA sequencing. Thirty four different mutations were identified by a mutation detection rate of 100%. IVS10-11G > A, p.P281L, R261Q, p.F39del and IVS11+1G > C were the most prevalent mutations with frequencies of 26.07%, 19.3%, 12.86%, 6.07 and 3.93%, respectively. All other mutations represented a relative frequency less than 3.5%. The data from this study provided a comprehensive spectrum of the PAH gene mutations which can facilitate carrier detection and prenatal diagnosis of PKU disease in the Iranian population. © 2018
International Journal Of Cancer Management (25384422)12(4)
Background: DNA topoisomerase II alpha (Top2-α) enzyme is an important target for many anticancer drugs. A variety of TOP2A genomic variants has been found associated with the development of drug resistance to this enzyme. Methods: Here, we have characterized 2 non-synonymous single nucleotide polymorphisms (nsSNPs) including rs762022284 and rs764177670 in TOP2A gene, which could affect its response to Amsacrine and Mitoxantrone as important inhibitors of the enzyme. The nsSNPs were genotyped in the Iranian population and the data were analyzed, using PLINK and PICcalc programs. Results: Genotyping data indicated the allele frequency of 0.30 (PIC = 0.42) and 0.05 (PIC = 0.09) for rs762022284 and rs764177670, respectively. Conclusions: The data suggested that the presence of rs762022284 and rs764177670 nsSNPs could affect Top2-α response to Amsacrine and Mitoxantrone, indicating the necessity of consideration of population-dependent genotypes in cancer chemotherapy, using these drugs. © 2019, Author(s).
Metabolic Brain Disease (15737365)33(4)pp. 1165-1173
In this study, we introduce a novel compound-primed multiplex ARMS PCR (CPMAP) for simultaneous detection of common PAH gene mutations. This approach was used successfully for simultaneous identification of six most common PAH gene mutations in 137 phenylketonuria patients in the Iranian population. A total of six normal and six mutant allele-specific primers and 4 common primers containing a tag sequence of 12 base pair at the 5ˊ-end were designed and used in two separate optimized multiplex ARMS reactions followed by hot-start PCR. The products were separated and visualized on 3% agarose gel. The CPMAP genotyping data were completely in accordance with the direct sequencing results. The CPMAP suggests a reliable, economical and rapid method for simultaneous detection of PAH point mutations using conventional PCR, which could be applied for diagnosis of other gene mutations. © 2018, Springer Science+Business Media, LLC, part of Springer Nature.
Neuroscience Letters (18727972)648pp. 66-69
Multiple sclerosis (MS) is one of the most common diseases of the central nervous system (CNS) in the Iranian population. To date, association of many genes with the prevalence and progression of the disease have been investigated. In the present study, the impact of rs1738074 single nucleotide polymorphism (SNP) in the TAGAP gene (TAGAP rs1738074) on the risk of MS was evaluated in a sample of the Iranian population. In a case control study, genotyping was performed on 300 patients and normal individuals. The data were analyzed using Pearson's chi-square test. The results showed a significant difference in the SNP frequency between case and control groups (p-value = 0.049). The genotype frequencies of TT, TC and CC in patients were 10.67%, 51.33% and 38%, respectively, and in normal individuals were 20.66%, 42.67% and 36.67%, respectively. The results showed a significant difference in the genotype frequency of T/T between the patient and control groups (p < 0.05). Interestingly, individuals with T/T genotype were estimated to be less susceptible to MS ((p-value = 0.025), Fisher's exact test), odd ratio was 2.18 (controls versus MS patients) with 95% CI: 1.137–4.187. The results suggested that TAGAP rs1738074 polymorphism could be considered as a risk factor in the prevalence of MS in the Iranian population. © 2017 Elsevier B.V.
Molecular Biology Research Communications (2322181X)6(1)pp. 1-11
Stem cell factor (SCF) is a critical protein with key roles in the cell such as hematopoiesis, gametogenesis and melanogenesis. In the present study a comparative analysis on nucleotide sequences of SCF was performed in Humanoids using bioinformatics tools including NCBI-BLAST, MEGA6, and JBrowse. Our analysis of nucleotide sequences to find closely evolved organisms with high similarity by NCBI-BLAST tools and MEGA6 showed that human and Chimpanzee (Pan troglodytes) were placed into the same cluster. By using JBrowse, we found that SCF in Neanderthal had a single copy number similar to modern human and partly conserved nucleotide sequences. Together, the results approved the gene flow and genetics similarity of SCF among human and P. troglodytes. This may suggest that during evolution, SCF gene transferred partly intact either on the basis of sequence or function from the same ancestors to P. troglodytes, the ancient human like Neanderthal, and then to the modern human.
Journal of Molecular Graphics and Modelling (10933263)75pp. 287-293
Crizotinib is an efficient antineoplastic drug for treatment of non-small cell lung carcinoma (NSCLC), which is identified as an anaplastic lymphoma kinase (ALK) inhibitor. F1174V is a recently identified acquired point mutation relating to the Crizotinib resistance in NSCLC patients. The mechanism of Crizotinib resistance relating to F1174V mutation as a non-active site mutation remains unclear. In this study, the molecular dynamic simulation was used to investigate the possible mechanisms by which F1174V mutation may affect the structure and activity of ALK kinase domain. The results suggested that F1174V mutation could cause two important secondary structure alterations, which led to the local conformational change in ALK kinase domain. This causes more positive free energy in the mutant complex in comparison with the wild-type one. In addition, our structural analyses illustrated that F1174V mutation could result in some important interactions, which represent the key characteristics of the ALK active conformation. This study provided a molecular mechanism for ALK Crizotinib resistance caused by F1174V mutation,which could facilitate designing more efficient drugs. © 2017
Journal of Babol University of Medical Sciences (15614107)19(6)pp. 42-49
BACKGROUND AND OBJECTIVE: Phenylketonuria (PKU) is the most prevalent inborn error of amino acid metabolism in the world and its incidence rate is increasing in Iran. The aim of the present study was to assess the efficiency of HRM technique as a fast and suitable method in identifying common mutations of phenylalanine hydroxylase gene including IVS10-11G>A and P281L in order to improve the early detection of the disease to prevent the occurrence of it. METHODS: In this case-control study20 DNA samples including one sample with IVS10-11G>A mutation, one sample with P281L mutation and 18 control samples were extracted from peripheral blood collected in Medical Genetic Center of Isfahan and were genotyped with HRM technique. To validate the mutations, the mutant samples were genotyped using sequencing. Bioinformatic analyses were used for determining structural and functional effects of P281L mutation on the of PAH protein. FINDINGS: HRM analysis identified IVS10-11G>A and P281L mutations with a sensitivity and specificity of 100% and the mutant and normal samples differentiated well in normalized and difference plots. Bioinformatic analyses demonstrated instability and pathological effects of PAH protein containing P281L mutation. CONCLUSION: HRM is a simple and fast technique detecting the two IVS10-11G>A and P281L mutations with 100% sensitivity and specificity. © 2017, Babol University of Medical Sciences. All rights reserved.
Genes and Genomics (19769571)39(4)pp. 433-443
Leber’s congenital amaurosis (LCA) is considered as one of the main causes of congenital blindness. In view of the genetically heterogeneous nature of the disease, indirect diagnosis using linkage analysis has proven to be useful in molecular diagnosis procedure. Mutations in AIPL1 gene are one of the leading causes of LCA. In the present study, the application of three single nucleotide polymorphic (SNP) markers related to the gene, including rs7212734, rs11658369 and rs8066853 was evaluated for the first time in the Iranian population. The markers were genotyped using tetra-primer ARMS PCR in 154 unrelated healthy individuals. Haplotype frequency and other characteristics of the markers were examined by using the GENEPOP, PowerMarker and Cubic Exact Solution software. The data indicated the presence of six different haplotypes in the Iranian population. Among them, three haplotypes showed high informativeness with frequencies ≥0.05. Three unrelated Iranian LCA families were analyzed. The p.W278* mutation in exon 6 of AIPL1 was found in all the families using APEX microarray chips. SNP analysis for the families showed the conservation of T−A−A haplotype linked to p.W278* mutation. All families bearing the mutation were from the Isfahan province and a founder effect was suggested in this population. Prenatal diagnosis using the markers led to the successful prediction of the fetus genotypes in all the at risk pregnancies and showed that rs7212734, rs11658369 and rs8066853 can be considered as the three informative markers for linkage analysis in carrier detection and molecular diagnosis of LCA in the Iranian population. © 2017, The Genetics Society of Korea and Springer-Science and Media.
Current Cancer Drug Targets (15680096)17(7)pp. 657-668
Background: DNA topoisomerase II-α (Top2-α), an essential enzyme for the management of DNA during replication, transcription, recombination, and chromatin remodeling, is one of the most important anticancer targets. Numerous molecules have been designed as Top2-α inhibitors. However, several studies have shown that polymorphisms and mutations in Top2 have conferred resistance to most of these anticancer drugs. The aim of this study was to computationally examine the mechanisms by which genomic variations in Top2-α could affect its resistance to Amsacrine and Mitoxantrone as important inhibitors of the enzyme. Results: The results showed that variants K529E, R568H, R568G and T530M could affect Top2-α inhibition by Amsacrine causing possible drug-resistant. Moreover, R487K, and Y481C variants could change the response of the enzyme to Mitoxantrone. Conclusion: These results could facilitate the prediction and development of more effective drugs for Top2-α variants, making the cancer chemotherapy more effective. © 2017 Bentham Science Publishers.
Moafi, A.,
Vallian, R.,
Vallian borujeni, S.,
Rahgozar, S.,
Torfenajad, M.,
Moafi, H. Journal of Medical Screening (09691413)24(1)pp. 1-5
Objective: To evaluate the repercussions of recent changes to the cut-offs used in the first screening step of the pre-marital screening programme for thalassaemia prevention in Iran. Methods: The profiles of 984 subjects referred to a genetic laboratory, and the tests of 242 parents of children with thalassaemia major were assessed for red blood cell (RBC) indices, haemoglobin (Hb) A2 levels and results of Hb electrophoresis. Results: Of 407 suspected thalassaemia minor (STM) cases, 18 proved positive for thalassaemia minor on molecular analysis (18/407, confidence interval 2.6–6.9%). If the revised screening cut-offs had been used to determine who would undergo molecular analysis, two of these cases would not have been identified. Only 4.4% of suspected cases with lower than normal RBC indices (mean corpuscular volume <80 fl and mean corpuscular Hb <27 pg) and HbA2 (<3.5%) were diagnosed with thalassaemia minor. Conclusion: The thalassaemia major prevention programme is performed in two separate steps. One step involves the screening of subjects and identification of b-thalassaemia minor, suspected cases for thalassaemia minor (STM), and normal subject groups. The other step concerns the identification of thalassaemia minor in the STM group. Changing the cut-offs at the first screening step does not result in significant improvement from an economic view, and is associated with significant risk at the second screening step. © The Author(s) 2016.
Iranian Journal Of Basic Medical Sciences (20083874)20(8)pp. 880-885
Objective(s): Mutations in the UGT1A1 gene are responsible for hyperbilirubinemia syndromes including Crigler-Najjar type 1 and 2 and Gilbert syndrome. In view of the genetic heterogeneity and involvement of large numbers of the disease causing mutations, the application of polymorphic markers in the UGTA1 gene could be useful in molecular diagnosis of the disease. Materials and Methods: In the present study, two polymorphic markers including rs4148326 and rs4124874 in the UGT1A1 gene region were characterized. The markers were selected using bioinformatics analysis of the UGT1A1 gene region and genotyped in 212 unrelated healthy individuals and 13 family trios in the Iranian population using Tetra-Primer ARMS PCR technique. The allele frequency and population status of the alleles were estimated using GENEPOP, FBAT, PowerMarker and Arlequin software. Results: The results indicated that in the case of rs4148326 marker, allele frequency for T and C allele was 66.04% and 33.96%, respectively. For rs4124874 marker, allele frequency for G and T alleles was 39.4% and 60.6%, respectively. The values of heterozygosity index for the markers examined were 64.1 for rs4148326 and 72.1 for rs4124874, respectively. The haplotype estimation analysis of the markers resulted in three informative haplotypes with frequencies ≥0.05. Moreover, the results suggested the presence of linkage disequilibrium between two markers. Conclusion: Altogether, the data suggested that rs4148326 and rs4124874 could be introduced as informative markers for molecular diagnosis of Crigler-Najjar type 1 and 2 and Gilbert syndrome in the Iranian population. © 2017, Mashhad University of Medical Sciences. All rights reserved.
Karbalaie, K.,
Vallian borujeni, S.,
Lachinani, L.,
Tanhaei, S.,
Baharvand, H.,
Nasr-esfahani, M.H. Iranian Journal Of Biotechnology (23222921)14(3)pp. 109-116
Background: Promyelocytic leukemia protein (PML) is a tumor suppressor protein that is involved in myeloid cell differentiation in response to retinoic acid (RA). In addition, RA acts as a natural morphogen in neural development. Objectives: This study aimed to examine PML gene expression in different stages of in vitro neural differentiation of NT2 cells, and to investigate the possible role of PML in pluripotency and/or neural development. Materials and Methods: RA was used as a neural inducer for in vitro neural differentiation of NT2 cells. During this process PML mRNA and protein levels were assessed by quantitative real time RT-PCR (QRT-PCR) and Immunoblotting, respectively. Furthermore bisulfite sequencing PCR (BSP) was used to assess PML promoter methylation in NT2 cells and NT2 derived neuronal precursor cells (NT2.NPCs). Results: QRT-PCR results showed that, PML had maximum expression with significant differences in NT2 derived neuronal precursor cells relative to NT2 cells and NT2 derived neural cells (NT2.NCs). Numerous isoforms of PML with different intensities appeared in immunoblots of pluripotent NT2 cells, NT2.NPCs, and NT2.NCs. Furthermore, the methylation of the PML promoter in NT2.NCs was 2.6 percent lower than NT2 cell. Conclusions: The observed differences in PML expression in different cellular stages possibly could be attributed to the fact that PML in each developmental state might be involved in different cell signaling machinery and different functions. The appearance of different PML isoforms with more intensity in neural progenitor cells; may suggest apossible role for this protein in neural development. © 2016, Kowsar Medical Publishing Company. All rights reserved.
Meta Gene (22145400)7pp. 16-19
Spinal muscular atrophy (SMA) is a degenerative neuromuscular disease associated with progressive symmetric weakness and atrophy of the limb muscles. In view of the involvement of numerous point mutations and deletions associated with the disease, the application of polymorphic markers flanking the SMA critical region could be valuable in molecular diagnosis of the disease. In the present study, D5S351 and D5S1414 polymorphic markers located at the SMA critical region in the Iranian populations were characterized. Genotyping of the markers indicated the presence of six and nine different alleles for D5S351 and D5S1414, respectively. Haplotype frequency estimation in 25 trios families and 75 unrelated individuals indicated the presence of six informative haplotypes with frequency higher than 0.05 in the studied population. Furthermore, the D' coefficient and the χ2 value for D5S351 and D5S1414 markers revealed the presence of linkage disequilibrium between the two markers in the Iranians. These data suggested that D5S351 and D5S1414 could be suggested as informative markers for linkage analysis and molecular diagnosis of SMA in the Iranian population. © 2015.
Genetics in the Third Millennium (24237159)13(4)
PML (Promyelotic Leukemia) protein is a tumor suppressor protein encoded by the PML gene. This protein is known as one of the main components of PML nuclear bodies (PML-NBs) in the nucleolus. The number and morphology of these structures varies in different cells. Due to alternative splicing, post-translational modifications, and interactions with different partners, this protein has acquired different localizations and functions in the cell. Most studies have focused on the role of this protein in tumor suppression and myeloid differentiation. In this article, we review the role of this protein in cellular activity, and its importance in stem cell characteristics and neural development. © 2016, Iranian Neurogenetics Society. All rights reserved.
Medicinal Chemistry Research (10542523)25(6)pp. 1250-1259
Topoisomerases are enzymes that resolve winding problem of DNA during cellular processes. Because of essential roles of these enzymes in maintenance of cell function, topoisomerases are important targets for cancer chemotherapy. To date, several topoisomerase inhibitors have been introduced and applied as drugs in the treatment of cancer. Topoisomerase II α (Top2-α), a subclass of topoisomerase II enzymes, functions as the target for several anticancer agents and a variety of mutations in this protein have been associated with the development of drug resistance. Mitoxantrone and Amsacrine are among two important inhibitors of Top2 enzymes used in cancer chemotherapy. In this study, we used computational methods to analysis interactions between these compounds and Top2-α in order to identify the most important residues involved in the enzyme inhibition. In order to obtain reliable results, several docking studies have been performed on the human Top2-β to reproduce binding modes which are observed in the crystal form of Top2-β complexed with Mitoxantrone and Amsacrine. Since human Top2-β is the closest homologue to Top2-α, same docking parameters have been used for docking of Top2-α with mentioned drugs. The data also showed that the main residues involved in the interaction between Top2-α and Mitoxantrone were Lys489, Asp504, Glu506, Gly488, Ile490 and Leu491. For Top2α-Amsacrine complex, the interaction was mainly through Arg487, Glu506, Gly488, Lys489, Asn504 and Ala505. These findings clarify the mechanisms of action for these drugs and may facilitate future drug development and cancer treatment. © 2016, Springer Science+Business Media New York.
Gene (18790038)570(2)pp. 180-184
Fragile X syndrome, which is caused by mutation in the FMR1 gene region, is one of the most prevalent forms of mental retardation. Direct diagnosis of the disease is based on PCR and southern blot analysis, but because of technical problems, use of polymorphic DNA markers can be helpful for carrier detection and prenatal diagnosis in families with an affected individual. The polymorphic markers usually show a population-based haplotype frequency and heterozygosity. In the present study, genotyping and analysis of haplotype frequency of three microsatellite markers including DXS998, DXS548 and FRAXAC1 at the FMR1 gene region were carried out in 140 unrelated healthy women and 26 families from the Iranian population. The data indicated the presence of a novel allele for DXS998 in the Iranian population. Estimation of haplotype frequency using Arlequin program showed 50 different DXS998-DXS548-FRAXAC1 haplotypes for the input data of 5, 7 and 4 alleles, respectively. Among these haplotypes five of them showed relatively high frequencies (≥ 0.05). Analysis of linkage disequilibrium (LD) for the unrelated individuals using the PowerMarker computer program, showed that this haplotype combination can be an informative haplotype for linkage analysis in carrier detection and possible molecular diagnosis of fragile X in the Iranian population. © 2015 Elsevier B.V.
Iranian Journal Of Basic Medical Sciences (20083874)18(6)pp. 571-575
Objective(s): Iran is considered as one of the high-prevalence areas for β -thalassemia with a rate of about 10% carrier frequency. Molecular diagnosis of the disease is performed both by direct sequencing and indirectly by the use of polymorphic markers present in the beta globin gene cluster. However, to date there is no reliable information on the application of the markers in the Iranian population. Here we report the results of an extended molecular analysis of five RFLP markers, Xmnl, HindlUA, HindlUG, Rsal and Hinfl, located within the β-globin gene cluster region in four subpopulations of Iran. Materials and Methods: A total of 552 blood samples taken from the Iranian subpopulations including Isfahan, Chaharmahal-O-Bakhtiari, Khuzestan and Hormozgan were genotyped using PCR-RFLP and sequencing. The allele frequency, the expected and observed heterozygosity, and Shannons information index (I) of these markers were calculated. Results: Distribution of the allele frequencies for XmnI, HindIIIA, HindIIIG, Rsal and HinfI polymorphic markers did not differ significantly among the subpopulations examined. Overall observed heterozygosity ranged from 0.1706 for HindlIIIA to 0.4484 for RsaI. The Shannon index was <1 for all the polymorphic markers in the populations studied. The data indicated that heterozygosity of these markers was low in the Iranian population. Conclusion: The results suggested that genotyping of these markers is not informative enough once used as single markers for prenatal diagnosis and carrier detection of β-thalassemia in the Iranian population. However, haplotyping of these markers may provide more useful data in linkage analysis and prenatal diagnosis as well as carrier detections for β-thalassemia in Iranians. © 2015, Mashhad University of Medical Sciences. All rights reserved.
Genetic Testing and Molecular Biomarkers (19450265)19(3)pp. 156-161
Objectives: Genotyping of single-nucleotide polymorphisms (SNPs) has been applied in various genetic contexts. Tetra-primer amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) is reported as a prominent assay for SNP genotyping. However, there were published data that may question the reliability of this method on some occasions, in addition to a laborious and time-consuming procedure of the optimization step. In the current study, a new SNP genotyping method named modified tetra-primer ARMS (MTPA) PCR was developed based on tetra-primer ARMS PCR. Design and Methods: The modified method has two improvements in its instruction, including equalization of outer primer and inner primer strength by additional mismatch in outer primers, and consideration of equal annealing temperature of specific fragments more than melting temperature of primers. Advantageously, a new computer software was provided for designing primers based on novel concepts. Results: The usual tetra-primer ARMS PCR has a laborious process for optimization. In nonoptimal PCR programs, identification of the accurate genotype was found to be very difficult. However, in MTPA PCR, equalization of the amplicons and primer strength leads to increasing specificity and convenience of genotyping, which was validated by sequencing. Conclusions: In the MTPA PCR technique, a new mismatch at -2 positions of outer primers and equal annealing temperature improve the genotyping procedure. Together, the introduced method could be suggested as a powerful tool for genotyping single-nucleotide mutations and polymorphisms. © Mary Ann Liebert, Inc. 2015.
Genetics in the Third Millennium (24237159)13(3)pp. 4034-4039
Promyelocytic leukemia (PML) is one of the major proteins in the PML nuclear bodies (PML-NBs). Retinoic acid (RA) exerts its tumor growth suppressor activity and terminal myeloid differentiation of granulocyte/ monocyte progenitor (GMP) cells throught PML-NBs in the RA pathway. RA acts as a natural morphogen during posterior patterning in embryo neural development. This study was aimed to define the possible PML involvemet in RA dependent neural development of human embryonic stem cells (hESCs). The expression level of PML at both mRNA and protein level was assessed by the quantitative real time RT-PCR (q-RT-PCR) and western blotting, respectively. The mRNA data showed that PML had maximum expression in pluripotent hESCs and this expression clearly decreased in neural precursor cells (hNPCs) and neural cells (hNCs). No significant differences existed between the two latter cell types at mRNA level, while protein assay showed numerous protein bands with different intensities in hESCs, hNPCs, and hNCs. The data suggest that PML may have an important role in cellular pluripotency, and PML might be necessary in cellular pluripotency and nervous system development. © 2015, Iranian Neurogenetics Society. All rights reserved.
Computational Biology and Chemistry (14769271)51pp. 57-62
Aspirin (ASA) is a commonly used nonsteroidal anti-inflammatory drug (NSAID), which exerts its therapeutic effects through inhibition of cyclooxygenase (COX) isoform 2 (COX-2), while the inhibition of COX-1 by ASA leads to apparent side effects. In the present study, the relationship between COX-1 non-synonymous single nucleotide polymorphisms (nsSNPs) and aspirin related side effects was investigated. The functional impacts of 37 nsSNPs on aspirin inhibition potency of COX-1 with COX-1/aspirin molecular docking were computationally analyzed, and each SNP was scored based on DOCK Amber score. The data predicted that 22 nsSNPs could reduce COX-1 inhibition, while 15 nsSNPs showed increasing inhibition level in comparison to the regular COX-1 protein. In order to perform a comparing state, the Amber scores for two Arg119 mutants (R119A and R119Q) were also calculated. Moreover, among nsSNP variants, rs117122585 represented the closest Amber score to R119A mutant. A separate docking computation validated the score and represented a new binding position for ASA that acetyl group was located within the distance of 3.86 Å from Ser529 OH group. This could predict an associated loss of activity of ASA through this nsSNP variant. Our data represent a computational sub-population pattern for aspirin COX-1 related side effects, and provide basis for further research on COX-1/ASA interaction. © 2014 Elsevier Ltd.
Genetics in the Third Millennium (24237159)11(4)pp. 3260-3269
Leber Congenital Amaurosis (LCA) is the most severe form of inherited retinopathy in humans. LCA is genetically heterogeneous and inherited in an autosomal recessive manner. Aryl hydrocarbon receptor interacting protein-like 1 (AIPL1) gene mutations have been identified to cause the most clinically severe forms of the disease. Direct sequencing is usually used to detect point mutations and other sequence variations in the gene, which is expensive and time consuming. Alternatively, linkage analysis of polymorphic markers such as single nucleotide polymorphism (SNP) has been used in heterozygous carrier detection and prenatal diagnosis of the disease in families with an affected individual. A large number of SNP markers have been introduced in the AIPL1 gene region in the electronic databases. In the present study, the characteristics of rs11658369 as an informative marker located in intron 2 of AIPL1 gene region was investigated. Genotyping was carried out by tetra-primer ARMS PCR technique in 154 unrelated healthy individuals from Isfahan, Iran, using newly designed primers. Estimation of allelic frequency, heterozygosity rate, and presence of Hardy Weinberg Equilibrium (HWE) was performed using GenePop website and the amount of polymorphism information content (PIC) was computed by Power Marker software for the marker. The results indicated 23.2% minor allele frequency (MAF), 38.56% heterozygosity rate and 29.29% PIC for rs11658369 marker in the Isfahan population. Moreover, analysis of Hardy-Weinberg Equilibrium showed the presence of equilibrium for this marker in the mentioned population. In total, according to the results of this study (MAF>0.2 and PIC>0.25), rs11658369 can be considered as a highly informative SNP marker for molecular diagnosis of AIPL1 related LCA by indirect genetic analysis in the Isfahan population as a representative sample of the Iranian population. © 2014, Iranian Neurogenetics Society. All rights reserved.
Genetic Testing and Molecular Biomarkers (19450265)18(12)pp. 820-825
Background and Aims: SLC26A4 gene mutations are the second currently identifiable genetic cause of autosomal recessive nonsyndromic hearing loss after GJB2 mutations. Because of the extensive size of the SLC26A4 gene and the variety of mutations, indirect diagnosis using linkage analysis has been suggested. Therefore, in this investigation three potential short tandem repeat (STR) markers related to this region including D7S2420, D7S496, and D7S2459 were selected for further analysis. Methods: The characteristics and haplotype frequency of the markers were examined for the first time in five ethnic groups of the Iranian population including Fars, Azari, Turkmen, Gilaki, and Arab using the polymerase chain reaction followed by fluorescent capillary electrophoresis. Results were analyzed by GeneMarker HID Human STR Identity, GenePop, Microsatellite tools, PowerMarker 3.25, and Arlequin 3.5 software.
Genetics in the Third Millennium (24237159)12(3)pp. 3692-3703
Infertility is often defined as the condition in which a couple cannot conceive after one year of unprotected intercourse. Male specific factors account for about 30-40% of infertility. In 40-50% of infertility cases, female specific factors are the cause, while the remaining 10-30% are either attributed to both male and female specific factors or are unexplained. Several factors are involved in male infertility that can be divided into genetic, epigenetic, and non-genetic factors. Spermatogenic Failure (SgF) accounts for more than half of male infertility cases. Microdeletions of Yq chromosome are the most significant recognized cause of SgF. Up to 15% of SgF is related to known Y chromosomal deletions. Extensive physical, functional, and genetic analyses of the Y chromosome have now identified three AZF regions (AZFa, AZFb and AZFc), which encode spermatogenic genes. Y chromosome microdeletions, depending on the type of deletion, result in the loss of spermatogenic genes. The detection of Y chromosome microdeletions is performed by known STS (Sequence Tag Sites) markers for these regions. Besides, a new hybridization method (Suspension Array Technology) is recently used for rapid and efficient detection of these deletions. In the present study, the association between Y chromosome microdeletions and male infertility, molecular mechanisms involved in microdeletions, molecular detection of Y chromosome microdeletions and some treatment strategies have been reviewed. © 2014, Iranian Neurogenetics Society. All rights reserved.
Human Immunology (01988859)74(8)pp. 965-969
Association between HLA-DRB1 and a large number of diseases such as multiple sclerosis, type I diabetes and rheumatoid arthritis has been demonstrated. In the present study, we attempted to identify and characterize potential microsatellites in the HLA-DRB1 gene region to find specific markers for genotyping and linkage analysis of this gene. The microsatellites including M2_3_22, M2_2_36, D6S2878, D6S2805, D6S2879 and D6S2880 were selected from microsatellite resource in the Major Histocompatibility Complex database (dbMHC). In silico analysis showed that only M2_3_22 was specific for HLA-DRB1. Moreover, our findings revealed some more accurate characteristics of the other investigated microsatellites. M2_3_22 existed as a single copy in all the MHC haplotype sequences and was located next to HLA-DRB1. Therefore, a new set of primers compatible with all the last published MHC haplotype sequences were designed and used to amplify M2_3_22 in 164 DNA samples obtained from unrelated Iranian individuals. M2_3_22 was successfully amplified in all DNA samples and three different alleles were identified. This locus was found in the Hardy-Weinberg equilibrium (P> 0.05) in the studied population. Together, the findings suggested that M2_3_22 could be introduced as a specific locus in the HLA-DRB1 gene region for linkage analysis and disease association studies. © 2013 American Society for Histocompatibility and Immunogenetics.
Gene (18790038)512(1)pp. 123-126
The application of polymorphic markers in construction of phylogenetic trees has been documented. Five polymorphic markers located in the PAH gene region including PAH- BglII, PAH- PvuII(A), PAH- EcoRI, PAH- MspI and PAH-STR were selected for analysis of phylogenetic relationships of the Iranians with 15 other populations of the world. The lowest genetic distance was observed between the Iranians and populations residing in Adygei (an ethnic group of the Russian Caucasus), Russia and Druze (a Middle Eastern group). However, East Asian populations including Han, Japanese and Cambodians, Khmer or the Oceanians (Melanesian, Nasioi) showed high genetic distance with the Iranians. The data suggested that the Iranians might have relatively close evolutionary history with the populations residing in Russia rather than East Asian populations. This study provided the first new molecular insight into the evolutionary history of the Iranian population. © 2012 Elsevier B.V.
Medical Oncology (13570560)29(1)pp. 5-9
In the present study, we examined the association of different alleles of MICA gene with the risk of breast cancer development in Iranian population. Our data showed a significant relationship between longer alleles, alleles with 9-and 6-GCT repeat of MICA gene, and a higher risk of developing breast cancer according to the age of onset. The data indicated a 6-fold increase for developing breast cancer in patients carrying the allele with 6-GCT repeat after age 50 (OR = 5.8333, 95% CI: 1.2976-26/2236, P = 0.0172). In addition, patients carrying longer alleles in their genotype (6/6, 6/9, and 9/9 genotypes) were found significantly at higher risk of developing breast cancer than control individuals (OR = 5.6, P = 0.0038, 95% CI: 1.6578-18.9166). In contrast, alleles with short GCT repeat of 4 and 5.1 showed to play a role in reducing the risk of breast cancer (OR = 0.79, P = 0.3643 and 95%CI: 0.4743-1.3157).Womenwith allele 4 were found twofold more protected against breast cancer (OR = 0.4597, 95% CI: 0.2164-0.9765, P = 0.0401). The results suggested that women with genotypes with 9-and 6-GCT repeat alleles of MICA gene could be consideredmore potent to develop breast cancer especially at higher age. © 2011 Springer Science+Business Media, LLC.
Iranian Journal Of Public Health (22516085)41(5)pp. 97-104
Background: Genetic diversity of three polymorphic markers in the phenylalanine hydroxylase (PAH) gene region including PvuII (a), PAHSTR and MspI were investigated. Methods: Unrelated individuals (n=139) from the Iranian populations were genotyped using primers specific to PAH gene markers including PvuII(a), MspI and PAHSTR. The amplified products for PvuII(a), MspI were digested using the appropriate restriction enzymes and separated on 1.5% agarose. The PAHSTR alleles were identified using polyacrylamide gel electrophoresis followed by silver staining. The exact size of the STR alleles was determined by sequencing. The allele frequency and population status of the alleles were estimated using PHASE, FBAT and GENEPOP software. Results: The estimated degree of heterozygosity for PAHSTR, MspI and PvuII (a) was 66%, 56% and 58%, respectively. The haplotype estimation analysis of the markers resulted in nine informative haplotypes with frequencies ≥5%. Moreover, the results obtained from Ewens-Watterson test for neutrality suggested that the markers were under balancing selection in the Iranian population. Conclusion: These findings suggested the presence of genetic diversity at these three markers in the PAH gene region. Therefore, the markers could be considered as functional markers for linkage analysis of the PAH gene mutations in the Iranian families with the PKU disease.
Molecular Biology Reports (03014851)39(12)pp. 11187-11199
The estimation of genetic distance between populations could improve our viewpoint about human migration and its genetic origin. In this study, we used allele frequency data of 12 polymorphic markers on 250 individuals (500 alleles) from the Iranian population to estimate genetic distance between the Iranians and other world populations. The phylogenetic trees for three different sets of allele frequency data were constructed. Our results revealed the genetic similarity between the Iranians and European populations. The lowest genetic distance was observed between the Iranians and some populations reside in Russia. Furthermore, the high genetic distance was observed between the Iranians and East Asian populations. The data suggested that the Iranians might have relatively close evolutionary history with Europeans, but historically independent from East Asian populations. The evaluation of genetic distance between Indians populations and Iranians was also performed. The Indian groups showed low genetic distance with others, but high genetic distance with the Iranians. This study could provide a new insight into the evolutionary history of the Iranian population.
Molecular Biology Reports (03014851)38(5)pp. 3395-3399
The variable number of tandem repeat (VNTR) marker located at the 30-end of the phenylalanine hydroxylase (PAH) gene, PAHVNTR marker, is commonly used in carrier detection and prenatal diagnosis of the PKU disease. During the molecular diagnosis of the disease, an artifact band associated with the PAHVTNR marker was frequently observed. Analysis of genotyping data from nine trios families indicated that in heterozygote individuals, the observed stutter (artifact) bands at PAHVNTR marker were minor bands with one repeat sequence shorter than the upper main bands. More investigations using sequencing revealed that the artifact band was associated with VNTRs including seven or higher core repeats. In statistical analysis, 75% of the studied heterozygote individuals represented PCR artifact band. The presence of the artifact band associated with PAHVNTR marker highlights a serious alarm risk of possible technical misdiagnosis in the application of the PAHVNTR marker in carrier detection and prenatal diagnosis of the PKU disease. © Springer Science+Business Media B.V. 2010.
Tissue Antigens (13990039)78(1)pp. 8-10
Medical Oncology (13570560)28(2)pp. 420-423
PTEN/MMAC1/TEP1 encodes a tumor suppressor protein, which regulates cell cycle progression, translation, and apoptosis by blocking the activation of Akt/PKB. The loss of PTEN function increases cell survival and induces tumor invasion. In this study, PTEN promoter status and its correlation with genetic and pathologic parameters were analyzed in genomic DNA from Iranian patients with breast cancer. DNA methylation patterns in the CpG islands were determined by a methylation-specific PCR (MSP) assay. PTEN promoter methylation was found to be present in 37 of 53(70%) tumor tissues and none in 20 normal counterparts. Moreover, promoter methylation was found in patients with heterozygote mutation in the PTEN gene. The pathological history of cancerous tissue sections showed that PTEN gene could be inactivated at the stages III and IV in sporadic breast cancer. These findings suggested that promoter hypermethylation of PTEN might contribute to the progression of sporadic breast cancer in human. © 2010 Springer Science+Business Media, LLC.
Cellular and Molecular Neurobiology (02724340)31(5)pp. 749-754
Non-syndromic sensorineural hearing loss (NSHL) represents the most common cause of hearing loss in the Iranian patients. In view of the large numbers of mutations identified in GJB2, mutations analysis of the gene has been time-consuming and cost-ineffective. Alternatively, molecular markers that are highly linked to the GJB2 gene have proven to be useful in carrier detection and prenatal diagnosis of NSHL families. These markers usually show a population-dependent-based haplotype frequency. However, to date, no information on the genotyping and frequency of the markers is present for the Iranian population. In this study, genotyping and analysis of the haplotype frequency of three markers, including BanI, D13S141, and D13S175, at the GJB2 region were investigated. The haplotype frequency was estimated using PHASE program. The input data contained two alleles (+ and -) for BanI, four alleles for D13S141, and seven alleles for D13S175. Among the 42 possible haplotypes examined, four haplotypes showed relatively high frequencies (≥5%). Therefore, a combination of BanI/D13S141/D13S175 could be suggested as an informative haplotype for possible carrier detection and prenatal diagnosis of NSHL in the Iranian population. © 2011 Springer Science+Business Media, LLC.
Iranian Journal Of Biotechnology (23222921)9(3)pp. 163-172
Phenylketonuria (PKU) is the most common autosomal recessive disorder of amino acid metabolism. The disease is caused mainly by mutations in the phenylalanine hydroxylase (PAH) gene, encoding phenylalanine hydroxylase (PAH) enzyme. The PAH enzyme deficiency results in the elevation of phenylalanine in the blood, which may cause severe irreversible mental retardation in the affected individuals. More than 500 different disease causing mutations have been identified in the PAH gene. Direct and indirect molecular approaches have been developed for carrier detection and prenatal diagnosis of PKU disease. Population distribution of the PAH gene mutations and the PKU disease varies in different countries. In view of relatively high prevalence of the disease in Iranian population, investigations toward the elucidation of molecular aspects of the disease were required. In the present article, clinical and molecular basis of the PKU disease, with emphasis on the studies performed in Iranian population, were reviewed.
Tissue Antigens (13990039)76(1)pp. 60-63
Genomewide screen analysis has shown the close association of the human leukocyte antigen (HLA)-DRB1 region with susceptibility to multiple sclerosis and a number of autoimmune diseases. Using bioinformatics software, several potential short tandem repeat (STR) markers have been introduced in this region in the major histocompatibility complex data base (dbMHC). In this study, the identity and characteristics of two putative STR markers, D6S2879 and D6S2806, in this region were examined in Iranian population. The loci were genotyped in 85 individuals using polymerase chain reaction followed by polyacrylamide gel electrophoresis and sequencing. Analysis of the allelic frequency showed the presence of six and four alleles for D6S2806 and D6S2879, respectively. Analysis of deviations from Hardy-Weinberg equilibrium (HWE) showed that D6S2806 was in equilibrium (P > 0.05). However, the D6S2879 locus showed a significant deviation from HWE (P < 0.05). Therefore, the D6S2806 locus could be suggested as a marker for linkage analysis and disease-susceptibility investigations in the MHC-DRB1 gene region. © 2010 John Wiley & Sons A/S.
Applied Biochemistry and Biotechnology (02732289)160(3)pp. 927-931
The single-strand conformation polymorphism (SSCP), accompanied by sequencing, is a useful methods for identifying mutations in a DNA fragment. In this study, we have developed a modified SSCP with the aid of sodium bisulfite treatment. The corresponding PCR products for exon 3 of Hb gene were sequenced and samples with homozygote and heterozygote single nucleotide substitutions were identified. The PCR products were treated with sodium bisulfite, which deaminates all the cytosine residues. The reaction mixture was then analyzed on non-denaturing polyacrylamide gels. The modified method, which is called deaminated SSCP (DSSCP), was applied successfully in analysis of mutations in the beta-globin gene at positions relevant to codon 6. DSSCP is a very effective and reproducible method providing clear results that are easy to interpret without the involvement of radioactivity. © 2009 Humana Press.
Psychiatry Research (01651781)170(2-3)pp. 271-272
The impact of a single nucleotide polymorphism (SNP) in the CD24 gene on the risk and progression of multiple sclerosis (MS) was investigated in the Iranian population. Our data revealed that the susceptibility and the progression of MS in individuals with the CD24V/V genotype were greater than in those with the CD24A/V and CD24A/A genotypes. © 2008 Elsevier Ireland Ltd. All rights reserved.
American Biotechnology Laboratory (07493223)27(4)pp. 10-11
International Journal of Human Genetics (discontinued) (09723757)9(2)pp. 115-121
Deficiency in phenylalanine hydroxylase (PAH) is the main molecular characteristic of phenylketonuria (PKU). So far over 500 different mutations in the PAH gene have been identified as associated with the disease. Mutation analysis of the PAH gene is a time-consuming and cost-effective procedure. Therefore, molecular markers which are highly linked to the PAH gene, have been used in carrier detection and prenatal diagnosis in PKU families. These markers show a population dependent based haplotype frequency. In the present study, the haplotype frequency of three markers including BglII, EcoRI and VNTR, at the PAH gene region were investigated in Iranian population. Nine different alleles for VNTR marker with 3, 5, 6, 7, 8, 9, 10, 11 and 13 core repeats were detected. Alleles 4 and 13 were found specific to the Iranian population. The haplotype frequency was calculated using FBAT, PHASE and Arlequin (haplotype frequency estimation computer programs). Among the 36 possible estimated haplotypes identified for 2 alleles (+ and -) for BglII and EcoRI; and nine alleles for VNTR, ten haplotypes showed relatively high frequency (e" 5%), based on the above programs. Therefore, a combination of BglII-EcoRI-VNTR could be suggested as an informative haplotype in performing carrier and prenatal diagnosis of the PAH gene mutations in Iranian population. © Kamla-Raj 2009.
Iranian Journal Of Biotechnology (23222921)7(3)pp. 137
In the present study, genotyping of six short tandem repeat (STR) loci including CSF1PO, D16S539, F13A01, F13B, LPL and HPRTB was performed on genomic DNA from 127 unrelated individuals from the Iranian province of Isfahan. The results indicated that the allele and genotype distributions were in accordance with Hardy-Weinberg expectations. The observed heterozygosity (Ho), expected heterozygosity (He) as well as forensic and paternity indices including power of discrimination (PD) and exclusion (PE), polymorphism information content (PIC), typical paternity index (PItypical) and probability of paternity (W) were determined for the examined STR alleles. In addition, genetic diversity index (GD) and population parameter (θ) were calculated for the six loci. The combined power of discrimination (Pd combined) and combined probability of exclusion (PE typical) were 0.9999998 and 0.999856 over the six loci, respectively. Together, the examined STR loci in this study have proven a relatively high genetic variation in Iranian population. The data could be used for construction of a forensic genetic database for Iranian population.
Journal Of Cancer Research And Clinical Oncology (01715216)135(8)pp. 991-996
Introduction: p16 INK4A is a tumor suppressor encoding the Cdk inhibitor protein, which acts to repress Cdk4/6 and pRb phosphorylation. p16 INK4A gene can be inactivated by a variety of events, including promoter hypermethylation. Materials and methods: To investigate the methylation status of the p16 INK4A gene in Iranian patients with breast carcinoma, promoter methylation was studied by methylation-specific PCR (MSP) and restriction enzyme-related PCR (REP). In addition, p16 INK4A promoter was analyzed by PCR-SSCP in order to detection of mutation and single nucleotide polymorphisms. Results: Analysis of 70 patients by MPS and REP showed hypermethylation of p16 INK4A promoter in 35.7% (25/70) and 40% (28/70) of samples, respectively. Comparison of the molecular data and pathological information of the samples suggested that p16 INK4A gene might be inactivated at the early stages in breast cancer. Conclusion: Therefore, it could be suggested that hypermethylation of p16 INK4A promoter is one of the epigenetic factors affecting the progress of sporadic breast carcinogenesis in Iranian patients. © 2008 Springer-Verlag.
Iranian Journal Of Public Health (22516085)38(4)pp. 136-139
Background: The haplotype phasing is more useful than genotyping markers independently at carrier detection and prenatal diagnosis of diseases. The PAH gene region contains several markers used in detection of PKU disease. In the present study, the efficiency of BglII-EcoRI-VNTR haplotype phasing in Iranian family trios was investigated. Then, this information was compared with those obtained for unrelated individuals. Methods: Blood samples were collected from 20 healthy family trios and 60 unrelated individuals. The genomic DNA was extracted by use of salting-out procedure. The two markers BglII and EcoRI were genotyped by use of PCR-RFLP. The genotype of VNTR marker was identified by use of PCR and electrophoresis. The genotyping data obtained from family trios was used to infer haplotype phase. We also compared this data with results obtained from a widely used method for haplotype frequency inference from unrelated individuals, the PHASE program. Results: The haplotype phase of all members was only ascertained at eight family trios. The comparison of this data with the results obtained by use of PHASE program showed that eight haplotypes [211, 221, 215, 216, 214, 121, 225 and 111] were informative haplotypes in Iranian population. Conclusion: Since diversity of BglII-EcoRI-VNTR haplotypes was high in Iranian population, haplotype phasing at family trios was difficult. The results of this study showed that the genotyping data obtained from family trios could not provide enough information for BglII-EcoRI-VNTR haplotype phasing at Iranian PKU families and the genotyping of other family members was necessary at most cases.
Iranian Journal Of Biotechnology (23222921)6(3)pp. 151-156
A single nucleotide polymorphism (SNP) in CD24 has been associated with multiple sclerosis (MS) in a population based study. This SNP results in the replacement of alanine (CD24A) by valine (CD24V) at amino acid 57 in the resulting polypeptide chain. In the current study, the genotyping of this SNP and its contribution to MS in 217 patients and 200 healthy individuals of an Iranian population was investigated. The correlation of the SNP alleles with the progression of the disease was determined using the expanded disability status scale (EDSS) and progression index (PI). The data revealed that individuals with the CD24V/V genotype showed a 2-fold increase in the relative risk of MS compared to patients with the CD24A/V (0.27) and CD24A/A (0.25) genotypes (P = 0.0193, Odds Ratio 2.4882, 95% CI: 1.416-4.3722). Moreover, the progression of the disease in patients with CD24V/V was much faster than other patients that were examined by ANOVA and the least significant difference (LSD) test. However, in the CD24V/V patients LSD analysis was statistically significant (p<0.05 and p<0.01). These results support the hypothesis that CD24 may function as a genetic modifier for susceptibility and progression of MS through the CD24V/V genotype.
Journal of Applied Sciences (discontinued) (18125654)8(17)pp. 3026-3031
In the present study, a new economic, rapid and inexpensive bacterial micro-assay for simultaneous detection and quantitative measurement of serum galactose was developed. Quantitative measurement of galactose in the serum of galactosemia patients is necessary to confirm the disease. Analysis of the standard curve showed a broad linearity range for galactose from 2 to 180 mg dL-1 with a regression equation of Y=0.013-0.083; R2=0.962. The advantage of the method is its ability to measure serum galactose quantitatively. The cost per sample is about 20-50 cents, which is much less than HPLC and enzymatic commercial kits. The method can be automated which is suitable for galactosemia neonatal and mass screening especially in developing countries in which funding is a limiting factor. © 2008 Asian Network for Scientific Information.
Biotechnology (discontinued) (1682296X)6(4)pp. 497-504
Poly (hydroxy alkanoic acid) (PHA) polymers are synthesized by numerous microorganisms. These compounds have similar properties to synthetic plastics with excellent biodegradability. Here we report identification and cloning of phaC1 and phaC2 genes of type II PHA synthase gene from the Iranian isolate of P. aeroginosaPTCC 1310 bacterium. The sequences of both genes were isolated using PCR amplification with specific primers and cloned into pTZ57R cloning vectors as pTZPHAC1 and pTZPHAC2. The correct sequence of the cloned genes was confirmed by restriction digestion followed by partial sequencing. The vectors could be used for future sub-cloning and expression analysis purposes. © 2007 Asian Network for Scientific Information.
Journal of Applied Genetics (12341983)47(1)pp. 79-83
Phenylketonuria is an inherited metabolic disease, which is characterized by increased level of serum phenylalanine (Phe). The quantitative measurement of Phe in the serum is necessary to confirm the disease, and to distinguish phenylketonuria from other forms of hyperphenylalaninemia. In this study, we report a rapid and inexpensive micro-assay for simultaneous detection and quantitative measurement of serum Phe in dry blood-spots. Analysis of the standard curve showed a broad linear Phe range of 120-1800 μmol L -1. Application of this method in conjunction with the standard Guthrie bacterial inhibition assay and high-pressure liquid chromatography in analyzing 34 samples from phenylketonuria patients and control samples produced comparable results, with the regression equation of Y= 0.994 + 0.996. The advantage of this method over the Guthrie bacterial inhibition assay is its ability to measure the serum Phe quantitatively without false positive results. The method was successfully applied to dried blood-spots as well as serum and whole blood samples. The cost per sample is about 20-50 US cents, which is much less than those of high-pressure liquid chromatography and enzymatic commercial kits. The method can be automated, which is suitable for neonatal and mass phenylketonuria screening, especially in developing countries, where funding is a limiting factor.
Genetic Resources and Crop Evolution (09259864)53(7)pp. 1477-1484
Microsatellite markers were used to analyse the biodiversity of 57 accessions of different subspecies and varieties of wild Aegilops tauschii (2n = 2x = 14; D genome) collected across the major areas where it grows in Iran. Levels of diversity were high, with numbers of alleles averaging 7.3 (ranging up to 12) and polymorphism information contents averaging 0.6591. One accession was notably more similar to two of the D genome in hexaploid wheats (Triticum aestivum) used as outgroups. Within the Ae. tauschii accessions, no markers were characteristic for taxa or geographical origin, suggesting high gene flow between the subspecies and varieties, although some groupings, which could be related to geographical origin, were evident. This survey demonstrates the high diversity present in wild goatgrass in Iran, and indicates that there is value in sampling for useful genes for wheat breeding. © 2006 Springer Science+Business Media, Inc.
Pakistan Journal of Biological Sciences (18125735)9(1)pp. 115-118
In this study, we isolated a mutant of Kluyveromyces marxianus resistance to glucose repression. To screen for depression mutants, the strains were treated with UV rays. Fifteen resistant mutant strains were isolated. The mutants were further screened for glucose-repression-resistant mutants in the presence of 2-deoxy-D-glucose, an analog to glucose and lactose as the sole carbon source. In this condition, one glucose-repression-resistant mutant was isolated. The enzyme activity in this mutant strain and the wild type strain was compared using different mediums containing 4% of each lactose and glucose and 2% glucose+2% lactose. The results demonstrated significant decreasing in glucose repression in the mutant strain as compared to the wild type. This mutant was unable to grow anaerobically on glucose in present of antimycin A, the property of rag1 mutants. This mutant is, therefore, capable of constitutive expression of β-galactosidase, which makes it suitable for industrial purposes. © 2006 Asian Network for Scientific Information.
Clinical Chemistry and Laboratory Medicine (14346621)44(1)pp. 76-79
Phenylketonuria is an inherited metabolic disease, which is characterized by an increased level of serum phenylalanine. Quantitative measurement of phenylalanine in the serum of phenylketonuria patients is necessary to confirm the disease, and to distinguish phenylketonuria from other forms of hyperphenylalaninemia. In this study, we report a rapid and inexpensive micro-assay for simultaneous detection and quantitative measurement of serum phenylalanine on dry blood-spots. Analysis of the standard curve showed a broad linear range for phenylalanine from 120 to 1800 μmol/L. Application of this method, the standard Guthrie bacterial inhibition assay and a high-performance liquid chromatography (HPLC) method for analysis of 34 samples from phenylketonuria patients and control samples produced comparable results, with the regression equation Y = 0.994X + 0.996. The advantage of this method over the Guthrie bacterial inhibition assay is its ability to measure serum phenylalanine quantitatively without false positive results. The method was successfully applied to dried blood-spots, serum and whole blood. The cost per sample is approximately 20-50 US cents, which is much less than for HPLC and commercial enzyme kits. The method can be automated, and is thus suitable for neonatal and mass screening for phenylketonuria, especially in developing countries where funding is a limiting factor. © 2006 by Walter de Gruyter.
Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis (13861964)526(1-2)pp. 45-52
Phenylalanine hydroxylase (PAH) deficiency is caused by mutations in the PAH gene (12q22-q24) resulting in a primary deficiency of the PAH enzyme activity, intolerance to the dietary intake of phenylalanine (Phe) and production of the phenylketonuria (PKU) disease. To date there have been no reports on the molecular analysis of PKU in Iranian population. In this study, the states of the PKU disease in terms of prevalence and mutation spectrum among patients reside in the institutions for mentally retarded in Isfahan was investigated. In the first step, 611 out of 1541 patients with PKU phenotype or severe mental retardation were screened for the PKU disease using the Guthrie bacterial inhibition assay (GBIA) followed by HPLC. Among the patients screened 34 (5.56%) were found positive with abnormal serum Phe of above 7mg/dl. In the next step, the presence of 18 common mutations of the PAH gene in 26 of the patients with classical PKU (serum Phe above 20mg/dl) was investigated, using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Of the 52 independent mutant alleles that were analyzed, 34 (65.38%) were genotyped showing 8 mutations as follows: R252W (15.38%), Q232Q (13.46%), R261Q (7.69%), delL364 (7.69%), IVS10-11g > a (5.77%), L333F (5.77%), V245V (5.77%) and S67P (3.85%). The results from this study may serve as a reference to analyze the PKU mutations in other part of Iran, and to establish diagnostic tests for carrier detection and prenatal diagnosis of the PKU disease in Iranian population. © 2003 Elsevier Science B.V. All rights reserved.
Obstetrics and Gynecology (00297844)91(3)pp. 319-323
Objective: To determine whether the mechanism for the retention of interstitial fluid in trisomy 21 fetuses presenting with nuchal translucency at 10-14 weeks' gestation is an alteration in the composition of collagen type VI, which is normally a triple helix formed of three single chains, α1, α2, and α3. The genes responsible for the α1 and α2 chains, COL6A1 and COL6A2, are located on chromosome 21 and therefore may be overexpressed in trisomy 21, whereas COL6A3 is located in chromosome 2. Methods: Skin tissue was obtained after termination of pregnancy at 11-16 weeks' gestation in five fetuses with trisomy 21 and five normal controls. Total RNA was extracted and the steady-state levels of COL6A1 and COL6A3 mRNA expression of the gene transcripts were determined. Additionally, the distribution of collagen type VI in the skin of trisomy 21 and normal fetuses was analyzed using an immunohistochemical method. Results: The ratio of the normalized densitometric scores for the mRNA expression of COL6A1 to COL6A3 in the skin of trisomy 21 fetuses was twice as high as in normal fetuses. Immunohistochemistry demonstrated that in trisomy 21 fetuses collagen type VI formed a dense network extending from the epidermal basement membrane to the subcutis, whereas in normal fetuses dense staining was confined to the upper region of the dermis. Conclusion: The distribution for collagen type VI is different from normal in the skin of trisomy 21 fetuses, and there is overexpression of COL6A1 compared with COL6A3.
Vallian borujeni, S.,
Gäken, J.A.,
Gingold, E.B.,
Kouzarides, T.,
Chang k.-s., K.,
Farzaneh, F. Oncogene (14765594)16(22)pp. 2843-2853
The growth and transformation suppressor function of promyelocytic leukemia (PML) protein are disrupted in acute promyelocytic leukemia (APL) as a result of its fusion to the RARα gene by t(15;17) translocation. There is significant sequence homology between the dimerization domain of PML and the Fos family of proteins, which imply that PML may be involved in AP-1 activity. Here we show that PML can cooperate with Fos to stimulate its AP-1-mediated transcriptional activity. Cotransfection of PML, with GAL4/Fos strongly induced Fos-mediated activation of GAL4-responsive reporters, indicating a functional interaction between Fos and PML in vivo. Deletion analysis of Fos and PML demonstrated that the intact C-terminal domain of Fos (containing the dimerization domain), and the RING-finger, B1 box and nuclear localization domains of PML are involved in the cooperative activity of Fos and PML. Immunoprecipitation and electrophoretic mobility shift assay showed that PML is associated with the AP-1 complex. PMLRARα was also found to enhance the transcriptional activity of GAL4/Fos. The addition of retinoic acid abrogated the PMLRARα, but not PML-induced stimulation of GAL4/Fos activity in a dose-dependent manner. This study demonstrated that PML is involved in the AP-1 complex and can modulate Fos-mediated transcriptional activity, which may contribute to its growth suppressor function.
Oncogene (14765594)16(14)pp. 1839-1849
Our previous studies demonstrated that the promyelocytic leukemia gene, PML which involved in the 15;17 translocation in acute promyelocytic leukemia (APL) is a growth and transformation suppressor. In this study, recombinant PML adenovirus, Ad-PML was constructed and used to infect human breast cancer cells in vitro and in vivo, the anti-oncogenic function of PML and its mechanism of growth suppressing effect in breast cancer cells were examined. We showed that Ad-PML effectively infected the MCF-7 and SK-BR-3 cells. A high level of PML protein was expressed within 24 h post-infection and a detectable level remained at day 16. Ad-PML significantly suppressed the growth rate, clonogenicity, and tumorigenicity of breast cancer cells. Intratumoral injections of MCF-7-induced tumors by high titer Ad-PML suppressed tumor growth in nude K mice by about 80%. The injection sites expressed high level of PML and associated with a massive apoptotic cell death. To elucidate the molecular mechanism of PML's growth suppressing function, we examined the effect of Ad-PML on cell cycle distribution in MCF-7 and SK-BR-3 cells. We found that Ad-PML infection caused a cell cycle arrest at the G1 phase. We further showed that G1 arrest of MCF-7 cells is associated with a significant decrease in cyclin D1 and CDK2. An increased expression of p53, p21 and cyclin E was found. The Rb protein became predominantly hypophosphorylated 48 h post-infection. These findings indicate that PML exerts its growth suppressing effects by modulating several key G1 regulatory proteins. Our study provides important insight into the mechanism of tumor suppressing function of PML and suggests a potential application of Ad-PML in human cancer gene therapy.
Molecular and Cellular Biology (02707306)18(12)pp. 7147-7156
The promyelocytic leukemia protein (PML) is a nuclear phosphoprotein with growth- and transformation-suppressing ability. Having previously shown it to be a transcriptional repressor of the epidermal growth factor receptor (EGFR) gene promoter, we have now shown that PML's repression of EGFR transcription is caused by inhibition of EGFR's Sp1-dependent activity. On functional analysis, the repressive effect of PML was mapped to a 150-hp element (the sequences between -150 and -16, relative to the ATG initiation site) of the promoter. Transient transfection assays with Sp1-negative Drosophila melanogaster SL2 cells showed that the transcription of this region was regulated by Sp1 and that the Sp1-dependent activity of the promoter was suppressed by PML in a dose-dependent manner. Coimmunoprecipitation and mammalian two-hybrid assays demonstrated that PML and Sp1 were associated in vivo. In vitro binding by means of the glutathione S-transferase (GST) pull-down assay, using the full-length and truncated GST- Sp1 proteins and in vitro-translated PML, showed that PML and Sp1 directly interacted and that the C-terminal (DNA-binding) region of Sp1 and the coiled-coil (dimerization) domain of PML were essential for this interaction. Analysis of the effects of PML on Sp1 DNA binding by electrophoretic mobility shift assay (EMSA) showed that PML could specifically disrupt the binding of Sp1 to DNA. Furthermore, cotransfection of PML specifically repressed Sp1, but not the E2F1-mediated activity of the dihydrofolate reductase promoter. Together, these data suggest that the association of PML and Sp1 represents a novel mechanism for negative regulation of EGFR and other Sp1 target promoters.
Carcinogenesis (01433334)18(11)pp. 2063-2069
Our previous studies demonstrated that PML is a growth suppressor that suppresses oncogenic transformation of NIH/3T3 cells and rat embryo fibroblasts. PML is a nuclear matrix-associated phosphoprotein whose expression is regulated during the cell cycle. Disruption of PML function by t(15;17) in acute promyelocytic leukemia (APL) plays a critical role in leukemogenesis. To further study the role of PML in the control of cell growth, we have stably overexpressed PML protein in the HeLa cell line. This overexpression of PML significantly reduced the growth rate of HeLa cells and suppressed anchorage-independent growth in soft agar. We consequently investigated several parameters correlated with cell growth and cell cycle progression. We found that, in comparison with the parental HeLa cells, HeLa/PML stable clones showed proportionally more cells in G1 phase, fewer cells in S phase and about the same number in G2/M phase. The HeLa/PML clones showed a significantly longer doubling time as a result of a lengthening of the G1 phase. No effect on apoptosis was found in HeLa cells overexpressing PML. This observation indicates that PML suppresses cell growth by increasing cell cycle duration as a result of G1 elongation. To further understand the mechanism of the effect of PML on HeLa cells, expression of cell cycle-related proteins in HeLa/PML and parental HeLa cells was analyzed. We found that Rb phosphorylation was significantly reduced in PML stable clones. Expression of cyclin E, Cdk2 and p27 proteins was also significantly reduced. These studies indicate that PML affects cell cycle progression by mediating expression of several key proteins that normally control cell cycle progression. These results further extend our current understanding of PML function in human cells and its important role in cell cycle regulation.
Vallian borujeni, S.,
Gäken, J.A.,
Trayner, I.D.,
Gingold, E.B.,
Kouzarides, T.,
Chang k.-s., K.,
Farzaneh, F. Experimental Cell Research (10902422)237(2)pp. 371-382
Acute promyelocytic leukemia is characterized by the presence of a t(15; 17) chromosomal translocation which results in the expression of a chimeric gene product, PMLRARα, consisting of an N-terminal-truncated retinoic acid receptor-α fused to a C-terminal-truncated PML. Several structural features, and regions of homology to known transcription factors, suggest that PML may be involved in the regulation of gene expression. In this study we have analyzed the transcriptional regulatory activity of PML using chimeric GAL4/PML constructs and GAL4-responsive reporter plasmids. The data presented demonstrate that PML, when fused to the DNA-binding domain of GAL4 (GAL4/PML), inhibits transcription from GAL4-responsive promoters. The magnitude of this repression is cell type and promoter dependent, and deletion studies show that the putative coiled-coil and part of the serine- rich regions of PML are required for this activity. These regions are also shown to be responsible for the repression of transcription activity from the EGFR promoter. The data presented also demonstrate that GAL4/PML can recruit PMLRARα resulting in the retinoid-inducible transcriptional activation of a GAL4-responsive promoter, a function dependent on the presence of the coiled- coil region of PMLRARα.
Gäken, J.A.,
Tavassoli, M.,
Gan, S.,
Vallian borujeni, S.,
Giddings, I.,
Darling, D.C.,
Galea-lauri, J.,
Thomas, M.G.,
Abedi, H.,
Schreiber, V. Journal of Virology (10985514)70(6)pp. 3992-4000
Integration of proviral DNA into the host cell genome is a characteristic feature of the retroviral life cycle. This process involves coordinate DNA strand break formation and rejoining reactions. The full details of the integration process are not yet fully understood. However, the endonuclease and DNA strand-joining activities of the virus-encoded integrase protein (IN) are thought to act in concert with other, as-yet-unidentified, endogenous nuclear components which are involved in the DNA repair process. The nuclear enzyme poly(ADP-ribose) polymerase (PARP), which is dependent on DNA strand breaks for its activity, is involved in the efficient repair of DNA strand breaks, and maintenance of genomic integrity, in nucleated eukaryotic cells. In the present work, we examine the possible involvement of PARP in the retroviral life cycle and demonstrate that inhibition of PARP activity, by any one of three independent mechanisms, blocks the infection of mammalian cells by recombinant retroviral vectors. This requirement for PARP activity appears to be restricted to processes involved in the integration of provirus into the host cell DNA. PARP inhibition does not affect viral entry into the host cell, reverse transcription of the viral RNA genome, postintegration synthesis of viral gene products, synthesis of the viral RNA genome, or the generation of infective virions. Therefore, efficient retroviral infection of mammalian cells is blocked by inhibition of PARP activity.
Arif, S.,
Vallian borujeni, S.,
Farzaneh, F.,
Zanone, M.M.,
James, S.L.,
Pietropaolo, M.,
Hettiarachchi, S.,
Vergani, D.,
Conway, G.S.,
Peakman, M. Journal of Clinical Endocrinology and Metabolism (0021972X)81(12)pp. 4439-4445
Autoantibodies directed against steroid hormone-producing cells (SCA) detectable by immunofluorescence are typically found in a small proportion of patients with premature ovarian failure (POF) as well as in other endocrine autoimmune diseases. The SCA pattern stains cells in the outer zones of the adrenal cortex, ovary, and testis. To identify the molecular target of SCA, an adrenal complementary DNA expression library was screened using SCA- positive serum, and the steroid enzyme 3β-hydroxysteroid dehydrogenase (3βHSD) was identified. Only 1 of 48 (2%) patients with idiopathic POF, not preselected for the presence of other autoimmune diseases, had SCA by immunofluorescence, whereas 10 of 48 (21%) had anti-3βHSD autoantibodies detectable by immunoblot using recombinant human enzyme compared with 6 of 115 (5%) control subjects (P = 0.002). Absorption of SCA-positive serum with recombinant human 3βHSD abolished the immunofluorescence pattern. We also examined the prevalence of anti-3βHSD autoantibodies in other endocrine autoimmune diseases. Two of 112 (2%) diabetic patients, but none of the thyroid or Addisonian patients, had SCA by immunofluorescence. Twenty-six (23%) diabetic subjects (P < 0.001 vs. controls), 3 of 18 thyroid patients (P > 0.05 vs. controls), and none of 4 Addisonian patients had anti-3βHSD autoantibodies. 3βHSD is the first steroid cell autoantigen defined at the molecular level to be associated with idiopathic POF occurring in the absence of other polyglandular diseases. Autoantibodies to 3βHSD in patients with other organ-specific autoimmune diseases indicate that the enzyme behaves as a typical target of polyendocrine autoimmunity. Anti-3βHSD autoantibodies in patients with POF may provide a marker of those subjects whose ovarian failure is autoimmune in origin and, as recent studies suggest, may be salvageable with glucocorticoid treatment.
