Document Detail


Molecular characterization of human Argonaute-containing ribonucleoprotein complexes and their bound target mRNAs.
MedLine Citation:
PMID:  18978028     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
microRNAs (miRNAs) regulate the expression of mRNAs in animals and plants through miRNA-containing ribonucleoprotein particles (RNPs). At the core of these miRNA silencing effector complexes are the Argonaute (AGO) proteins that bind miRNAs and mediate target mRNA recognition. We generated HEK293 cell lines stably expressing epitope-tagged human AGO proteins and other RNA silencing-related proteins and used these cells to purify miRNA-containing RNPs. Mass spectrometric analyses of the proteins associated with different AGO proteins revealed a common set of helicases and mRNA-binding proteins, among them the three trinucleotide repeat containing proteins 6 (TNRC6A,-B,-C). mRNA microarray analyses of these miRNA-associated RNPs revealed that AGO and TNRC6 proteins bind highly similar sets of transcripts enriched in binding sites for highly expressed endogenous miRNAs, indicating that the TNRC6 proteins are a component of the mRNA-targeting miRNA silencing complex. Together with the very similar proteomic composition of each AGO complex, this result suggests substantial functional redundancy within families of human AGO and TNRC6 proteins. Our results further demonstrate that we have developed an effective biochemical approach to identify physiologically relevant human miRNA targets.
Authors:
Markus Landthaler; Dimos Gaidatzis; Andrea Rothballer; Po Yu Chen; Steven Joseph Soll; Lana Dinic; Tolulope Ojo; Markus Hafner; Mihaela Zavolan; Thomas Tuschl
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-10-31
Journal Detail:
Title:  RNA (New York, N.Y.)     Volume:  14     ISSN:  1469-9001     ISO Abbreviation:  RNA     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-12-05     Completed Date:  2008-12-24     Revised Date:  2013-06-04    
Medline Journal Info:
Nlm Unique ID:  9509184     Medline TA:  RNA     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2580-96     Citation Subset:  IM    
Affiliation:
Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, New York 10065, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Eukaryotic Initiation Factors / metabolism*
Humans
Immunoprecipitation
MicroRNAs / metabolism
Proteome
RNA, Messenger / metabolism*
Ribonucleoproteins / metabolism
Ribosomes / metabolism
Transfection
Grant Support
ID/Acronym/Agency:
GM068476/GM/NIGMS NIH HHS; GM073047/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute
Chemical
Reg. No./Substance:
0/Eukaryotic Initiation Factors; 0/MIRN122 microRNA, human; 0/MicroRNAs; 0/Proteome; 0/RNA, Messenger; 0/Ribonucleoproteins
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