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首页> 外文期刊>Molecular and Cellular Biology >Members of the Large Maf Transcription Family Regulate Insulin Gene Transcription in Islet β Cells
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Members of the Large Maf Transcription Family Regulate Insulin Gene Transcription in Islet β Cells

机译：大黑手党转录家族成员调节胰岛β细胞中的胰岛素基因转录。

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The C1/RIPE3b1 (?118/?107 bp) binding factor regulates pancreatic-β-cell-specific and glucose-regulated transcription of the insulin gene. In the present study, the C1/RIPE3b1 activator from mouse βTC-3 cell nuclear extracts was purified by DNA affinity chromatography and two-dimensional gel electrophoresis. C1/RIPE3b1 binding activity was found in the roughly 46-kDa fraction at pH 7.0 and pH 4.5, and each contained N- and C-terminal peptides to mouse MafA as determined by peptide mass mapping and tandem spectrometry. MafA was detected in the C1/RIPE3b1 binding complex by using MafA peptide-specific antisera. In addition, MafA was shown to bind within the enhancer region (?340/?91 bp) of the endogenous insulin gene in βTC-3 cells in the chromatin immunoprecipitation assay. These results strongly suggested that MafA was the β-cell-enriched component of the RIPE3b1 activator. However, reverse transcription-PCR analysis demonstrated that mouse islets express not only MafA but also other members of the large Maf family, specifically c-Maf and MafB. Furthermore, immunohistochemical studies revealed that at least MafA and MafB were present within the nuclei of islet β cells and not within pancreas acinar cells. Because MafA, MafB, and c-Maf were each capable of specifically binding to and activating insulin C1 element-mediated expression, our results suggest that all of these factors play a role in islet β-cell function.
机译：C1 / RIPE3b1（？118 /？107 bp）结合因子调节胰腺β细胞特异性和葡萄糖调节的胰岛素基因转录。在本研究中，通过DNA亲和层析和二维凝胶电泳纯化了来自小鼠βTC-3细胞核提取物的C1 / RIPE3b1活化剂。 C1 / RIPE3b1结合活性在pH 7.0和pH 4.5的约46-kDa部分中发现，并且通过肽质谱图和串联质谱法测定，每个结合物都包含与小鼠MafA的N和C末端肽。通过使用MafA肽特异性抗血清，在C1 / RIPE3b1结合复合物中检测到了MafA。此外，在染色质免疫沉淀试验中，MafA在βTC-3细胞的内源性胰岛素基因的增强子区域（？340 /？91 bp）内结合。这些结果强烈表明，MafA是RIPE3b1激活剂的β细胞富集成分。然而，逆转录-PCR分析表明，小鼠胰岛不仅表达MafA，而且表达大Maf家族的其他成员，特别是c-Maf和MafB。此外，免疫组织化学研究表明，至少MafA和MafB存在于胰岛β细胞的细胞核内，而不存在于胰腺腺泡细胞内。由于MafA，MafB和c-Maf均能够特异性结合并激活胰岛素C1元素介导的表达，因此我们的结果表明，所有这些因素均在胰岛β细胞功能中起作用。

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