TY - JOUR
T1 - Argonaute proteins couple chromatin silencing to alternative splicing
AU - Ameyar-Zazoua, Maya
AU - Rachez, Christophe
AU - Souidi, Mouloud
AU - Robin, Philippe
AU - Fritsch, Lauriane
AU - Young, Robert
AU - Morozova, Nadya
AU - Fenouil, Romain
AU - Descostes, Nicolas
AU - Andrau, Jean Christophe
AU - Mathieu, Jacques
AU - Hamiche, Ali
AU - Ait-Si-Ali, Slimane
AU - Muchardt, Christian
AU - Batsché, Eric
AU - Harel-Bellan, Annick
N1 - Funding Information:
The authors thank A. Krainer (Cold Spring Harbor Laboratory) for the antibody to SRSF1 (anti-SRSF1) G. Meister (Regensburg University) for anti-AGO1 no. 4B8 and anti-AGO2 no. 11A9, Z. Mourelatos (University of Pennsylvania) for anti-AGO2 no. 2A8, A. Tarakhovsky (Rockefeller University), G. Hannon (Cold Spring Harbor Laboratory) and M. Otsuka (University of Tokyo) for the kind gift of knockout MEFs, A. Polesskaya (Centre National de la Recherche Scientifique) for generating the tagged AGO2 C2C12 cells, M. Zavolan and M. Khorshid (Basel University) for their help with bioinformatics, the Taplin Biological Mass Spectrometry Facility at Harvard Medical School for MS analysis, P. de la Grange (GenoSplice) for help with bioinformatics and J.B. Weitzman (University Paris Diderot), E. Allemand (Institut Pasteur) and L. Pritchard (Centre National de la Recherche Scientifique) for critical reading of the manuscript. This work was supported by the European Commission Sixth Framework Programme (Integrated Project Silencing RNAs: Organisers and Coordinators of Complexity in Eukaryotic Organisms (SIROCCO) contract number LSHG-CT-2006-037900, to A.H.-B.) and by the Agence Nationale de la Recherche (contract number ANR-11-BSV8-0013 to C.M., J.-C.A. and A.H.-B.).
PY - 2012/10
Y1 - 2012/10
N2 - Argonaute proteins play a major part in transcriptional gene silencing in many organisms, but their role in the nucleus of somatic mammalian cells remains elusive. Here, we have immunopurified human Argonaute-1 and Argonaute-2 (AGO1 and AGO2) chromatin-embedded proteins and found them associated with chromatin modifiers and, notably, with splicing factors. Using the CD44 gene as a model, we show that AGO1 and AGO2 facilitate spliceosome recruitment and modulate RNA polymerase II elongation rate, thereby affecting alternative splicing. Proper AGO1 and AGO2 recruitment to CD44 transcribed regions required the endonuclease Dicer and the chromobox protein HP1γ, and resulted in increased histone H3 lysine 9 methylation on variant exons. Our data thus uncover a new model for the regulation of alternative splicing, in which Argonaute proteins couple RNA polymerase II elongation to chromatin modification.
AB - Argonaute proteins play a major part in transcriptional gene silencing in many organisms, but their role in the nucleus of somatic mammalian cells remains elusive. Here, we have immunopurified human Argonaute-1 and Argonaute-2 (AGO1 and AGO2) chromatin-embedded proteins and found them associated with chromatin modifiers and, notably, with splicing factors. Using the CD44 gene as a model, we show that AGO1 and AGO2 facilitate spliceosome recruitment and modulate RNA polymerase II elongation rate, thereby affecting alternative splicing. Proper AGO1 and AGO2 recruitment to CD44 transcribed regions required the endonuclease Dicer and the chromobox protein HP1γ, and resulted in increased histone H3 lysine 9 methylation on variant exons. Our data thus uncover a new model for the regulation of alternative splicing, in which Argonaute proteins couple RNA polymerase II elongation to chromatin modification.
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U2 - 10.1038/nsmb.2373
DO - 10.1038/nsmb.2373
M3 - Article
C2 - 22961379
AN - SCOPUS:84867230570
SN - 1545-9993
VL - 19
SP - 998
EP - 1005
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 10
ER -