Hira-dependent histone H3.3 deposition facilitates PRC2 recruitment at developmental loci in ES cells.
Polycomb repressive complex 2 (PRC2) regulates gene expression during lineage specification through trimethylation of lysine 27 on histone H3 (H3K27me3). In Drosophila, polycomb binding sites are dynamic chromatin regions enriched with the histone variant H3.3. Here, we show that, in mouse embryonic stem cells (ESCs), H3.3 is required for proper establishment of H3K27me3 at the promoters of developmentally regulated genes. Upon H3.3 depletion, these promoters show reduced nucleosome turnover measured by deposition of de novo synthesized histones and reduced PRC2 occupancy. Further, we show H3.3-dependent interaction of PRC2 with the histone chaperone, Hira, and that Hira localization to chromatin requires H3.3. Our data demonstrate the importance of H3.3 in maintaining a chromatin landscape in ESCs that is important for proper gene regulation during differentiation. Moreover, our findings support the emerging notion that H3.3 has multiple functions in distinct genomic locations that are not always correlated with an "active" chromatin state.
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
Laura A
LA
Banaszynski
Howard Hughes Medical Institute, Ansary Stem Cell Institute, and Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, 10065 USA.
Duancheng
D
Wen
Genomics Resource Center, The Rockefeller University, New York, New York, 10065 USA.
Scott
S
Dewell
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
Sarah J
SJ
Whitcomb
Department of Genetics, Albert Einstein College of Medicine, Bronx, New York, 10461 USA.
Mingyan
M
Lin
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
Nichole
N
Diaz
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
Simon J
SJ
Elsässer
Molecular Medicine Unit, Institute of Child Health, London WC1N 1EH, UK.
Ariane
A
Chapgier
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
Aaron D
AD
Goldberg
The Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Isreal.
Eli
E
Canaani
Howard Hughes Medical Institute, Ansary Stem Cell Institute, and Department of Genetic Medicine, Weill Cornell Medical College, New York, New York, 10065 USA.
Shahin
S
Rafii
Department of Genetics, Albert Einstein College of Medicine, Bronx, New York, 10461 USA.|Departments of Neurology and Neuroscience, Albert Einstein College of Medicine, Bronx, New York, 10461 USA.
Deyou
D
Zheng
Laboratory of Chromatin Biology & Epigenetics, The Rockefeller University, New York, New York, 10065 USA.
C David
CD
Allis
D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism; D002454:Cell Differentiation; D002843:Chromatin / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D056488:Histone Chaperones / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D063151:Polycomb Repressive Complex 2 / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D015854:Up-Regulation
D018797:Cell Cycle Proteins; D002843:Chromatin; C484187:Hira protein, mouse; D056488:Histone Chaperones; D014157:Transcription Factors; D063151:Polycomb Repressive Complex 2; D012319:RNA Polymerase II