H19 inhibits RNA polymerase II-mediated transcription by disrupting the hnRNP U-actin complex
H19 inhibits RNA polymerase II-mediated transcription by disrupting the hnRNP U-actin complex
复制标题
H19 通过破坏 hnRNP U-肌动蛋白复合物来抑制 RNA 聚合酶 II 介导的转录
DOI：
10.1016/j.bbagen.2013.06.026
复制
发表时间：
2013-10-01
影响因子：
3
通讯作者：
Sun, Shu-han
中科院分区：
文献类型：
--
作者：
Bi, Hai-shan;Yang, Xiang-yue;Sun, Shu-han
背景：H19是最早被发现的，也是研究最多的长非编码RNA之一。据推测，H19对调节发育和疾病状况是必不可少的，它与许多类型的癌症发生有关。然而，这种保守的RNA的生物学功能和调控机制，特别是它对转录的影响，在很大程度上还不清楚。方法：我们通过RNA下拉、RNA免疫沉淀和缺失定位来鉴定与H19相关的蛋白质。结果：我们的研究进一步证实了H19与hnRNP U结合，这种相互作用位于H19的5‘882个核苷酸区域。此外，H19干扰hnRNP U与肌动蛋白之间的相互作用，从而抑制RNA聚合酶II(Pol II)C末端结构域(CTD)Ser5的磷酸化，从而阻止RNA Pol II介导的转录。结论：在本研究中，我们证明了H19通过破坏hnRNP U-肌动蛋白复合体来抑制RNA Pol II介导的转录。一般意义：这些数据表明H19在生物体内调节一般转录并发挥广泛的作用。(C)2013爱思唯尔B.V.保留所有权利。
Background: H19 was one of the earliest identified, and is the most studied, long noncoding RNAs. It is presumed that H19 is essential for regulating development and disease conditions, and it is associated with carcinogenesis for many types. However the biological function and regulatory mechanism of this conserved RNA, particularly with respect to its effect on transcription, remain largely unknown.Methods: We performed RNA pulldown, RNA immunoprecipitation and deletion mapping to identify the proteins that are associated with H19. In addition, we employed EU (5-ethynyl uridine) incorporation, immunoprecipitation and Western blotting to investigate the functional aspects of H19.Results: Our research further verifies that H19 is bound to hnRNP U, and this interaction is located within the 5' 882 nt region of H19. Moreover, H19 disrupts the interaction between hnRNP U and actin, which inhibits phosphorylation at Ser5 of the RNA polymerase II (Pol II) C-terminal domain (CTD), consequently preventing RNA Pol II-mediated transcription. We also showed that hnRNP U is essential for H19-mediated transcription repression.Conclusions: In this study, we demonstrate that H19 inhibits RNA Pol II-mediated transcription by disrupting the hnRNP U-actin complex.General significance: These data suggest that H19 regulates general transcription and exerts wide-ranging effects in organisms. (c) 2013 Elsevier B.V. All rights reserved.