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PMID: 17671087 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Let-7 microRNA-mediated mRNA deadenylation and translational repression in a mammalian cell-free system.

Genes & development ·Vol. 21 ·No. 15 ·2007-08-01 ·Pages 1857-62

Wakiyama M, Takimoto K, Ohara O, Yokoyama S

Abstract

MicroRNAs (miRNAs) are incorporated into miRNP complexes and regulate protein expression post-transcriptionally through binding to 3'-untranslated regions of target mRNAs. Here we describe a recapitulation of let-7 miRNA-mediated translational repression in a cell-free system, which was established with extracts prepared from HEK293F cells overexpressing miRNA pathway components. In this system, both the cap and poly(A) tail are required for the translational repression, and let-7 directs the deadenylation of target mRNAs. Our work suggests that let-7 miRNPs containing Argonaute and GW182 impair the synergistic enhancement of translation by the 5'-cap and 3'-poly(A) tail, resulting in translational repression.

MeSH Terms
Argonaute Proteins Autoantigens/genetics Base Sequence Cell Line Cell-Free System Eukaryotic Initiation Factor-2/genetics Gene Silencing Genes, Reporter Humans In Vitro Techniques Luciferases, Firefly/genetics MicroRNAs/genetics,metabolism Polyadenylation Protein Biosynthesis RNA Caps/chemistry,genetics,metabolism RNA, Messenger/chemistry,genetics,metabolism RNA, Small Interfering/genetics RNA-Binding Proteins
Chemicals
AGO2 protein, human Argonaute Proteins Autoantigens Eukaryotic Initiation Factor-2 MicroRNAs RNA Caps RNA, Messenger RNA, Small Interfering RNA-Binding Proteins TNRC6A protein, human mirnlet7 microRNA, human Luciferases, Firefly
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wakiyama Motoaki
RIKEN Genomic Sciences Center, Protein Research Group, Tsurumi, Yokohama 230-0045, Japan.
Takimoto Koji
Ohara Osamu
Yokoyama Shigeyuki
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1566707
Published
2007-08-01
Pages
1857-62
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1935024
Subset
IM
Corrections
ErratumIn
-
CommentIn
PMID: 17699746
External Links
PubMed source DOI: 10.1101/gad.1566707 PMC Full Text View journal details
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