Liganded retinoic acid X receptor α represses connexin 43 through a potential retinoic acid response element in the promoter region
Liganded retinoic acid X receptor α represses connexin 43 through a potential retinoic acid response element in the promoter region
复制标题
配体视黄酸 X 受体 α 通过启动子区域中潜在的视黄酸反应元件抑制连接蛋白 43
DOI：
10.1038/pr.2016.47
复制
发表时间：
2016-07-01
影响因子：
3.6
通讯作者：
Huang, Guoying
中科院分区：
文献类型：
--
作者：
Gu, Ruoyi;Xu, Jun;Huang, Guoying
前言：视黄酸X受体α（RXRa）和连接蛋白43（Cx43）在心脏发生中起着至关重要的作用。方法：采用荧光素酶转染法检测Cx43中视黄酸反应元件（RARE）的激活情况。电泳迁移率变动分析（EMSA）和染色质免疫沉淀（ChIP）证明RXR α可以直接与RARE序列结合。采用实时荧光定量聚合酶链反应（qRT-PCR）和蛋白质印迹法（western blotting）分析RXR α和Cx43在细胞水平的mRNA水平和蛋白水平。结果：本研究证实了RXR α和Cx43在细胞水平的基因表达呈负相关。有趣的是，在Cx43转录起始位点上游1，426至314个碱基对的区域中检测到功能性RARE。结论：RXR α通过与Cx43启动子中的RARE序列直接结合，负调控Cx43基因的转录和表达。在Cx43启动子区域中的RARE样序列可以作为视黄酸（RA）信号通路中的功能性RARE。
INTRODUCTION: Retinoic acid X receptor alpha (RXRa) and Connexin 43 (Cx43) both play a crucial role in cardiogenesis. However, little is known about the interplay mechanism between the RXRa and Cx43.METHODS:The activations of retinoic acid response element (RARE) in Cx43 were measured by luciferase transfection assay. Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) was performed to prove that RXRa can directly bind to the RARE sequence. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting were used to analyze the RXRa and Cx43 mRNA level and protein level in cells.RESULTS: In this study, we confirmed the negative association of the gene expression between the RXRa and Cx43 in the cell level., Interestingly, a functional RARE was detected in the region from 1,426 to 314 base pairs upstream from the transcriptional start site of Cx43. Moreover, we also prove that RXRa can directly bind to this RARE sequence in vitro and in vivo.CONCLUSIONS: RXRa negatively regulates the transcription and expression by directly binding to the RARE in the promoter of Cx43. The RARE-like sequence harbored in the Cx43 promoter region may serve as a functional RARE in the retinoic acid (RA) signaling pathway.