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microRNA-mediated messenger RNA deadenylation contributes to translational repression in mammalian cells
microRNA-mediated messenger RNA deadenylation contributes to translational repression in mammalian cells
- Source :
- PLoS ONE, Vol 4, Iss 8, p e6783 (2009), PLoS ONE
- Publication Year :
- 2009
- Publisher :
- Public Library of Science (PLoS), 2009.
-
Abstract
- Animal microRNAs (miRNAs) typically regulate gene expression by binding to partially complementary target sites in the 3' untranslated region (UTR) of messenger RNA (mRNA) reducing its translation and stability. They also commonly induce shortening of the mRNA 3' poly(A) tail, which contributes to their mRNA decay promoting function. The relationship between miRNA-mediated deadenylation and translational repression has been less clear. Using transfection of reporter constructs carrying three imperfectly matching let-7 target sites in the 3' UTR into mammalian cells we observe rapid target mRNA deadenylation that precedes measureable translational repression by endogenous let-7 miRNA. Depleting cells of the argonaute co-factors RCK or TNRC6A can impair let-7-mediated repression despite ongoing mRNA deadenylation, indicating that deadenylation alone is not sufficient to effect full repression. Nevertheless, the magnitude of translational repression by let-7 is diminished when the target reporter lacks a poly(A) tail. Employing an antisense strategy to block deadenylation of target mRNA with poly(A) tail also partially impairs translational repression. On the one hand, these experiments confirm that tail removal by deadenylation is not strictly required for translational repression. On the other hand they show directly that deadenylation can augment miRNA-mediated translational repression in mammalian cells beyond stimulating mRNA decay. Taken together with published work, these results suggest a dual role of deadenylation in miRNA function: it contributes to translational repression as well as mRNA decay and is thus critically involved in establishing the quantitatively appropriate physiological response to miRNAs.
- Subjects :
- Untranslated region
Small interfering RNA
Science
Biology
03 medical and health sciences
0302 clinical medicine
Molecular Biology/Translational Regulation
microRNA
Protein biosynthesis
Humans
RNA, Messenger
Psychological repression
DNA Primers
030304 developmental biology
0303 health sciences
Messenger RNA
Multidisciplinary
Base Sequence
Reverse Transcriptase Polymerase Chain Reaction
Translation (biology)
Argonaute
Molecular biology
Cell biology
MicroRNAs
Protein Biosynthesis
030220 oncology & carcinogenesis
Molecular Biology/Post-Translational Regulation of Gene Expression
Medicine
Molecular Biology/mRNA Stability
Molecular Biology/RNA-Protein Interactions
Poly A
Research Article
Molecular Biology/Translation Mechanisms
HeLa Cells
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 4
- Issue :
- 8
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....3c563593660c29a2befcacd07de7e588