Mammalian miRNA RISC Recruits CAF1 and PABP to Affect PABP-Dependent Deadenylation

Fabian, M. R., Mathonnet, G., Sundermeier, T., Mathys, H., Zipprich, J. T., Svitkin, Y. V., Rivas, F., Jinek, M., Wohlschlegel, J., Doudna, J. A., Chen, C. Y. A., Shyu, A. B., Yates Iii, J. R., Hannon, G. J., Filipowicz, W., Duchaine, T. F., Sonenberg, N. (2009) Mammalian miRNA RISC Recruits CAF1 and PABP to Affect PABP-Dependent Deadenylation. Molecular Cell, 35 (6). pp. 868-880. ISSN 10972765 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/19716330
DOI: 10.1016/j.molcel.2009.08.004

Abstract

MicroRNAs (miRNAs) inhibit mRNA expression in general by base pairing to the 3′UTR of target mRNAs and consequently inhibiting translation and/or initiating poly(A) tail deadenylation and mRNA destabilization. Here we examine the mechanism and kinetics of miRNA-mediated deadenylation in mouse Krebs-2 ascites extract. We demonstrate that miRNA-mediated mRNA deadenylation occurs subsequent to initial translational inhibition, indicating a two-step mechanism of miRNA action, which serves to consolidate repression. We show that a let-7 miRNA-loaded RNA-induced silencing complex (miRISC) interacts with the poly(A)-binding protein (PABP) and the CAF1 and CCR4 deadenylases. In addition, we demonstrate that miRNA-mediated deadenylation is dependent upon CAF1 activity and PABP, which serves as a bona fide miRNA coactivator. Importantly, we present evidence that GW182, a core component of the miRISC, directly interacts with PABP via its C-terminal region and that this interaction is required for miRNA-mediated deadenylation. © 2009 Elsevier Inc. All rights reserved.

Item Type: Paper
Uncontrolled Keywords: PROTEINS RNA Animals Ascites Autoantigens Binding Sites Carcinoma, Krebs 2 Cell-Free System Eukaryotic Initiation Factor-2 Eukaryotic Initiation Factor-4G Gene Silencing Hela Cells Humans Kinetics Mice MicroRNAs Poly(A)-Binding Proteins Protein Biosynthesis Protein Structure, Tertiary Receptors, CCR4 RNA Processing, Post-Transcriptional RNA Stability RNA, Messenger RNA-Induced Silencing Complex Transfection Mammalia
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > miRNA
CSHL Authors:
Communities: CSHL labs > Hannon lab
Depositing User: Matt Covey
Date: 2009
Date Deposited: 09 Jan 2013 16:43
Last Modified: 09 Jan 2013 16:43
PMCID: PMC2803087
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26438

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