A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.

H2A.Z is an essential histone variant that has been implicated to have multiple chromosomal functions. To understand how H2A.Z participates in such diverse activities, we sought to identify downstream effector proteins that are recruited to chromatin via H2A.Z. For this purpose, we developed a nucle...

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Autores principales:	Ryan Draker, Marlee K Ng, Elizabeth Sarcinella, Vladimir Ignatchenko, Thomas Kislinger, Peter Cheung
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Genetics QH426-470
Acceso en línea:	https://doaj.org/article/59175a5876e24bd7965f118470c9bc53
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spelling	oai:doaj.org-article:59175a5876e24bd7965f118470c9bc532021-11-18T06:20:34ZA combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.1553-73901553-740410.1371/journal.pgen.1003047https://doaj.org/article/59175a5876e24bd7965f118470c9bc532012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23144632/pdf/?tool=EBIhttps://doaj.org/toc/1553-7390https://doaj.org/toc/1553-7404H2A.Z is an essential histone variant that has been implicated to have multiple chromosomal functions. To understand how H2A.Z participates in such diverse activities, we sought to identify downstream effector proteins that are recruited to chromatin via H2A.Z. For this purpose, we developed a nucleosome purification method to isolate H2A.Z-containing nucleosomes from human cells and used mass spectrometry to identify the co-purified nuclear proteins. Through stringent filtering, we identified the top 21 candidates, many of which have conserved structural motifs that bind post-translationally modified histones. We further validated the biological significance of one such candidate, Brd2, which is a double-bromodomain-containing protein known to function in transcriptional activation. We found that Brd2's preference for H2A.Z nucleosomes is mediated through a combination of hyperacetylated H4 on these nucleosomes, as well as additional features on H2A.Z itself. In addition, comparison of nucleosomes containing either H2A.Z-1 or H2A.Z-2 isoforms showed that significantly more Brd2 co-purifies with the former, suggesting these two isoforms engage different downstream effector proteins. Consistent with these biochemical analyses, we found that Brd2 is recruited to AR-regulated genes in an H2A.Z-dependent manner and that chemical inhibition of Brd2 recruitment greatly inhibits AR-regulated gene expression. Taken together, we propose that Brd2 is a key downstream mediator that links H2A.Z and transcriptional activation of AR-regulated genes. Moreover, this study validates the approach of using proteomics to identify nucleosome-interacting proteins in order to elucidate downstream mechanistic functions associated with the histone variant H2A.Z.Ryan DrakerMarlee K NgElizabeth SarcinellaVladimir IgnatchenkoThomas KislingerPeter CheungPublic Library of Science (PLoS)articleGeneticsQH426-470ENPLoS Genetics, Vol 8, Iss 11, p e1003047 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Genetics QH426-470
spellingShingle	Genetics QH426-470 Ryan Draker Marlee K Ng Elizabeth Sarcinella Vladimir Ignatchenko Thomas Kislinger Peter Cheung A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
description	H2A.Z is an essential histone variant that has been implicated to have multiple chromosomal functions. To understand how H2A.Z participates in such diverse activities, we sought to identify downstream effector proteins that are recruited to chromatin via H2A.Z. For this purpose, we developed a nucleosome purification method to isolate H2A.Z-containing nucleosomes from human cells and used mass spectrometry to identify the co-purified nuclear proteins. Through stringent filtering, we identified the top 21 candidates, many of which have conserved structural motifs that bind post-translationally modified histones. We further validated the biological significance of one such candidate, Brd2, which is a double-bromodomain-containing protein known to function in transcriptional activation. We found that Brd2's preference for H2A.Z nucleosomes is mediated through a combination of hyperacetylated H4 on these nucleosomes, as well as additional features on H2A.Z itself. In addition, comparison of nucleosomes containing either H2A.Z-1 or H2A.Z-2 isoforms showed that significantly more Brd2 co-purifies with the former, suggesting these two isoforms engage different downstream effector proteins. Consistent with these biochemical analyses, we found that Brd2 is recruited to AR-regulated genes in an H2A.Z-dependent manner and that chemical inhibition of Brd2 recruitment greatly inhibits AR-regulated gene expression. Taken together, we propose that Brd2 is a key downstream mediator that links H2A.Z and transcriptional activation of AR-regulated genes. Moreover, this study validates the approach of using proteomics to identify nucleosome-interacting proteins in order to elucidate downstream mechanistic functions associated with the histone variant H2A.Z.
format	article
author	Ryan Draker Marlee K Ng Elizabeth Sarcinella Vladimir Ignatchenko Thomas Kislinger Peter Cheung
author_facet	Ryan Draker Marlee K Ng Elizabeth Sarcinella Vladimir Ignatchenko Thomas Kislinger Peter Cheung
author_sort	Ryan Draker
title	A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
title_short	A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
title_full	A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
title_fullStr	A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
title_full_unstemmed	A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.
title_sort	combination of h2a.z and h4 acetylation recruits brd2 to chromatin during transcriptional activation.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/59175a5876e24bd7965f118470c9bc53
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A combination of H2A.Z and H4 acetylation recruits Brd2 to chromatin during transcriptional activation.

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