STAT5a promotes the transcription of mature mmu-miR-135a in 3T3-L1 cells by binding to both miR-135a-1 and miR-135a-2 promoter elements
STAT5a promotes the transcription of mature mmu-miR-135a in 3T3-L1 cells by binding to both miR-135a-1 and miR-135a-2 promoter elements
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STAT5a 通过结合 miR-135a-1 和 miR-135a-2 启动子元件促进 3T3-L1 细胞中成熟 mmu-miR-135a 的转录
DOI：
10.1016/j.biocel.2016.06.003
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发表时间：
2016-08-01
影响因子：
4
通讯作者：
Jiang, Siwen
中科院分区：
文献类型：
--
作者：
Wei, Xiajie;Cheng, Xiaoyan;Jiang, Siwen
尽管对miR-135 a在生物过程中的作用进行了广泛的研究，但很少有人关注其转录的调控。我们以前报道过miR-135 a通过直接靶向腺瘤性结肠息肉病（APC）基因并激活经典的Wnt/β-连环蛋白信号通路来抑制3 T3-Li前脂肪细胞分化和脂肪形成，但是对miR-135 a的两种亚型（miR-135 a-1和miR-135 a-2）表达的调控元件仍然知之甚少。在这里，通过使用缺失分析，我们预测了两个结合位点miR-135 a-1和miR-135 a-2核心启动子内转录因子信号转导子和转录激活子5a（STAT 5a）（-874/-856和-2020/-2002）（-1128/-556和2264/-1773），随后的定点突变表明两个STAT 5a结合位点调节miR-135 a-1和miR-135 a-2启动子的活性。通过电泳迁移率变动分析（EMSA）和染色质免疫沉淀（ChIP）分析鉴定了STAT 5a与miR-135 a-1/2核心启动子在体外和细胞培养中的结合。STAT 5a的过表达和RNAi敲低表明该转录因子调节内源性miR-135 a的表达。STAT 5a和APC的表达时程表明它们之间存在潜在的负反馈。总之，本研究的总体结果表明，STAT 5a通过与miR-135 a-1和miR-135 a-2启动子元件结合来调节miR-135 a的转录，这些发现为miR-135 a在脂肪形成过程中的分子调控机制提供了新的见解。(C)2016爱思唯尔有限公司版权所有
Despite extensive research on the role of miR-135a in biological processes, very little attention has been paid to the regulation of its transcription. We have previously reported that miR-135a suppresses 3T3-Li preadipocyte differentiation and adipogenesis by directly targeting the adenomatous polyposis coli (APC) gene and activating the canonical Wnt/beta-catenin signaling pathway, but the regulatory elements that regulate the expression of the two isoforms of miR-135a (miR-135a-1 and miR-135a-2) remain poorly understood. Here, by using deletion analysis, we predicted two binding sites (-874/-856 and -2020/-2002) for the transcription factor Signal Transducers and Activators of Transcription 5a (STAT5a) within the core promoters of miR-135a-1 and miR-135a-2 (-1128/-556 and 2264/-1773), and the subsequent site-directed mutagenesis indicated that the two STAT5a binding sites regulated the activity of the miR-135a-1 and miR-135a-2 promoters. The binding of STAT5a to the miR-135a-1/2 core promoters in vitro and in cell culture was identified by electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) assays. Overexpression and RNAi knockdown of STAT5a showed that the transcription factor regulated the endogenous miR-135a expression. Additionally, The expression time frame of STAT5a and APC indicated a potential negative feedback between them. In sum, the overall results from this study indicate that STAT5a regulates miR-135a transcription by binding to both miR-135a-1 and miR135a-2 promoter elements and the findings provide novel insights into the molecular regulatory mechanisms of miR-135a during adipogenesis. (C) 2016 Elsevier Ltd. All rights reserved.