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)
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: | https://repository.cshl.edu/id/eprint/26438 |
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