Unknown,Transcriptomics,Genomics,Proteomics

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RNAi-independent role for Argonaute2 in CTCF/CP190 chromatin insulator function


ABSTRACT: Chromatin insulators and Polycomb group (PcG) complexes control nuclear organization to effect changes in gene expression. In Drosophila, RNA silencing pathways influence long range interactions mediated by PcG proteins and nuclear localization of the gypsy insulator; however, the underlying mechanisms are unknown. Here, we identify a singular requirement for Argonaute2 (AGO2) for the activity of the CCCTC-binding factor (CTCF)/Centrosomal protein 190 (CP190) dependent Fab-8 insulator. AGO2 and CP190 interact physically, and genome wide localization of AGO2 by chromatin immunoprecipitation and sequencing (ChIP-seq) reveals extensive colocalization of AGO2 with insulators and Polycomb Response Elements (PREs) but minimal overlap with regions of endogenous small interfering RNA (endo-siRNA) production. Finally, depletion of either CTCF or CP190 results in loss of AGO2 association with insulators, PREs, and other cis-regulatory regions. Our findings suggest that Dicer-independent recruitment of AGO2 to chromatin by insulator proteins promotes the definition of transcriptional domains throughout the genome. ChIP-seq of AGO2 in two Drosophila cell types (S2 and S3)

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Ryan Dale 

PROVIDER: E-GEOD-22623 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNAi-independent role for Argonaute2 in CTCF/CP190 chromatin insulator function.

Moshkovich Nellie N   Nisha Parul P   Boyle Patrick J PJ   Thompson Brandi A BA   Dale Ryan K RK   Lei Elissa P EP  

Genes & development 20110801 16


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 chrom  ...[more]

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