Expression of TNRC6 (GW182) Proteins Is Not Necessary for Gene Silencing by Fully Complementary RNA Duplexes
Expression of TNRC6 (GW182) Proteins Is Not Necessary for Gene Silencing by Fully Complementary RNA Duplexes
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DOI：
10.1089/nat.2019.0815
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发表时间：
2019-12-01
影响因子：
4
通讯作者：
Corey, David R.
中科院分区：
文献类型：
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作者：
Liu, Zhongtian;Johnson, Samantha T.;Corey, David R.
包含6(TNRC6)蛋白家族的三核苷酸重复序列是RNA干扰(RNAi)的核心成分，由三个平行序列(TNRC6A、TNRC6B和TNRC6C)组成。TNRC6类似物与核心RNAi因子ArgAerte(AGO)蛋白相关，并连接其与其他蛋白质的相互作用。我们获得了TNRC6A和TNRC6B单基因和双基因敲除细胞系，以研究TNRC6同源基因对RNAi的作用。我们发现，当双链RNA完全互补时，基因沉默不需要TNRC6蛋白。TNRC6的表达是microRNA调控所必需的。针对基因启动子的双重RNA的转录激活需要TNRC6A而不是TNRC6B的表达。相比之下，AGO2是所有三种基因表达途径所必需的。TNRC6A可以影响DICER在胞浆和胞核的定位，但三个TNRC6对AGO2的胞核定位都不是必需的。我们的数据表明，TNRC6对映体的作用在某些细节上是不同的，并且对于涉及完全互补的双链RNA的临床治疗沉默机制来说，TNRC6不是必需的。
The trinucleotide repeat containing 6 (TNRC6) family of proteins are core components of RNA interference (RNAi) and consist of three paralogs (TNRC6A, TNRC6B, and TNRC6C). The TNRC6 paralogs associate with argonaute (AGO) protein, the core RNAi factor, and bridge its interactions with other proteins. We obtained TNRC6A and TNRC6B single and double knockout cell lines to investigate how the TNRC6 paralogs contribute to RNAi. We found that TNRC6 proteins are not required for gene silencing when duplex RNAs are fully complementary. TNRC6 expression was necessary for regulation by a microRNA. TNRC6A, but not TNRC6B, expression was necessary for transcriptional activation by a duplex RNA targeting a gene promoter. By contrast, AGO2 is required for all three gene expression pathways. TNRC6A can affect the Dicer localization in cytoplasm versus the nucleus, but none of the three TNRC6 paralogs was necessary for nuclear localization of AGO2. Our data suggest that the roles of the TNRC6 paralogs differ in some details and that TNRC6 is not required for clinical therapeutic silencing mechanisms that involve fully complementary duplex RNAs.