Document Detail

RNAi-independent role for Argonaute2 in CTCF/CP190 chromatin insulator function.
MedLine Citation:
PMID:  21852534     Owner:  NLM     Status:  MEDLINE    
A major role of the RNAi pathway in Schizosaccharomyces pombe is to nucleate heterochromatin, but it remains unclear whether this mechanism is conserved. To address this question in Drosophila, we performed genome-wide localization of Argonaute2 (AGO2) by chromatin immunoprecipitation (ChIP)-seq in two different embryonic cell lines and found that AGO2 localizes to euchromatin but not heterochromatin. This localization pattern is further supported by immunofluorescence staining of polytene chromosomes and cell lines, and these studies also indicate that a substantial fraction of AGO2 resides in the nucleus. Intriguingly, AGO2 colocalizes extensively with CTCF/CP190 chromatin insulators but not with genomic regions corresponding to endogenous siRNA production. Moreover, AGO2, but not its catalytic activity or Dicer-2, is required for CTCF/CP190-dependent Fab-8 insulator function. AGO2 interacts physically with CTCF and CP190, and depletion of either CTCF or CP190 results in genome-wide loss of AGO2 chromatin association. Finally, mutation of CTCF, CP190, or AGO2 leads to reduction of chromosomal looping interactions, thereby altering gene expression. We propose that RNAi-independent recruitment of AGO2 to chromatin by insulator proteins promotes the definition of transcriptional domains throughout the genome.
Nellie Moshkovich; Parul Nisha; Patrick J Boyle; Brandi A Thompson; Ryan K Dale; Elissa P Lei
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Intramural    
Journal Detail:
Title:  Genes & development     Volume:  25     ISSN:  1549-5477     ISO Abbreviation:  Genes Dev.     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-08-19     Completed Date:  2011-12-07     Revised Date:  2013-06-28    
Medline Journal Info:
Nlm Unique ID:  8711660     Medline TA:  Genes Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1686-701     Citation Subset:  IM    
Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.
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MeSH Terms
Animals, Genetically Modified
Argonaute Proteins / genetics,  metabolism*
Binding Sites / genetics
Blotting, Western
Cell Line
Chromatin Immunoprecipitation
Cluster Analysis
Drosophila Proteins / genetics,  metabolism*
Euchromatin / genetics,  metabolism
Fluorescent Antibody Technique, Indirect
Gene Expression Profiling
Genome, Insect / genetics
Homeodomain Proteins / genetics,  metabolism
Microtubule-Associated Proteins / genetics,  metabolism*
Nuclear Proteins / genetics,  metabolism*
Promoter Regions, Genetic / genetics
Protein Binding
RNA Interference
RNA, Small Interfering / genetics,  metabolism
Repressor Proteins / genetics,  metabolism*
Reverse Transcriptase Polymerase Chain Reaction
Reg. No./Substance:
0/AGO2 protein, Drosophila; 0/Abd-B proteins, Drosophila; 0/Argonaute Proteins; 0/CP190 protein, Drosophila; 0/CTCF protein, Drosophila; 0/Drosophila Proteins; 0/Euchromatin; 0/Homeodomain Proteins; 0/Microtubule-Associated Proteins; 0/Nuclear Proteins; 0/RNA, Small Interfering; 0/Repressor Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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