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Nucleic Acids Res. 2017 Jun 20;45(11):6894-6910. doi: 10.1093/nar/gkx307.

WIG1 is crucial for AGO2-mediated ACOT7 mRNA silencing via miRNA-dependent and -independent mechanisms.

Author information

1
Department of Molecular Medicine, Medical Research Center, Inha University College of Medicine, Incheon 22212, Korea.
2
Medical Research Center, Inha University College of Medicine, Incheon 22212, Korea.
3
Laboratory of Genetics, National Institute on Aging-Intramural Research Program, NIH, Baltimore, MD 21224, USA.
4
Department of Computer Science and Engineering, Inha University, Incheon 22212, Korea.
5
Department of Molecular Biology, Dankook University, Yongin 16890, Korea.
6
Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.
7
Department of Microbiology, Inha University College of Medicine, Incheon 22212, Korea.
8
Division of Life Sciences, Korea University, Seoul 02841, Korea.

Abstract

RNA-binding proteins (RBPs) are involved in mRNA splicing, maturation, transport, translation, storage and turnover. Here, we identified ACOT7 mRNA as a novel target of human WIG1. ACOT7 mRNA decay was triggered by the microRNA miR-9 in a WIG1-dependent manner via classic recruitment of Argonaute 2 (AGO2). Interestingly, AGO2 was also recruited to ACOT7 mRNA in a WIG1-dependent manner in the absence of miR-9, which indicates an alternative model whereby WIG1 controls AGO2-mediated gene silencing. The WIG1-AGO2 complex attenuated translation initiation via an interaction with translation initiation factor 5B (eIF5B). These results were confirmed using a WIG1 tethering system based on the MS2 bacteriophage coat protein and a reporter construct containing an MS2-binding site, and by immunoprecipitation of WIG1 and detection of WIG1-associated proteins using liquid chromatography-tandem mass spectrometry. We also identified WIG1-binding motifs using photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation analyses. Altogether, our data indicate that WIG1 governs the miRNA-dependent and the miRNA-independent recruitment of AGO2 to lower the stability of and suppress the translation of ACOT7 mRNA.

PMID:
28472401
PMCID:
PMC5499809
DOI:
10.1093/nar/gkx307
[Indexed for MEDLINE]
Free PMC Article

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