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GAS41 interacts with transcription factor AP-2β and stimulates AP-2β-mediated transactivation

机译:GAS41与转录因子AP-2β相互作用并刺激AP-2β介导的反式激活

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摘要

Transcription factor AP-2 regulates transcription of a number of genes involving mammalian development, differentiation and carcinogenesis. Recent studies have shown that interaction partners can modulate the transcriptional activity of AP-2 over the downstream targets. In this study, we reported the identification of GAS41 as an interaction partner of AP-2β. We documented the interaction both in vivo by co-immunoprecipitation as well as in vitro through glutathione S-transferase (GST) pull-down assays. We also showed that the two proteins are co-localized in the nuclei of mammalian cells. We further mapped the interaction domains between the two proteins to the C-termini of both AP-2β and GAS41, respectively. Furthermore, we have identified three critical residues of GAS41 that are important for the interaction between the two proteins. In addition, by transient co-expression experiments using reporter containing three AP-2 consensus binding sites in the promoter region, we found that GAS41 stimulates the transcriptional activity of AP-2β over the reporter. Finally, electrophoretic mobility shift assay (EMSA) suggested that GAS41 enhances the DNA-binding activity of AP-2β. Our data provide evidence for a novel cellular function of GAS41 as a transcriptional co-activator for AP-2β.
机译:转录因子AP-2调节涉及哺乳动物发育,分化和致癌作用的许多基因的转录。最近的研究表明,相互作用伙伴可以调节AP-2在下游靶标上的转录活性。在这项研究中,我们报告了GAS41作为AP-2β相互作用伴侣的鉴定。我们记录了体内通过共免疫沉淀以及通过谷胱甘肽S-转移酶(GST)下拉测定法在体外的相互作用。我们还表明,这两种蛋白共定位在哺乳动物细胞的细胞核中。我们进一步将两个蛋白质之间的相互作用域映射到AP-2β和GAS41的C末端。此外,我们已经确定了GAS41的三个关键残基,它们对于两种蛋白质之间的相互作用至关重要。此外,通过使用在启动子区域包含三个AP-2共有结合位点的报告基因进行瞬时共表达实验,我们发现GAS41会刺激AP-2β超过报告基因的转录活性。最后,电泳迁移率迁移分析(EMSA)表明GAS41增强了AP-2β的DNA结合活性。我们的数据提供了GAS41作为AP-2β转录共激活因子的新型细胞功能的证据。

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