Genomics

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298

H2A.Z inheritance during the cell cycle and its impact on promoter organization and dynamics [ChIP-seq]


ABSTRACT: While it has been clearly established that well positioned H2A.Z-containing nucleosomes flank the nucleosome depleted region (NDR) at the transcriptional start site (TSS) of active mammalian genes, how this chromatin-based information is transmitted through the cell cycle is unknown. We show here that in trophoblast stem (TS) cells, the level of H2A.Z at promoters decreases during S phase coinciding with homotypic (H2A.Z/H2A.Z) nucleosomes flanking the TSS becoming heterotypic (H2A.Z/H2A). Surprisingly, these nucleosomes remain heterotypic at M phase. At the TSS, we identify an unstable heterotypic H2A.Z-containing nucleosome in G1 which, strikingly, is lost following DNA replication. These dynamic changes in H2A.Z at the TSS mirror a global expansion of the NDR at S and M which, unexpectedly, is unrelated to transcriptional activity. Coincident with the loss of H2A.Z at promoters, it is targeted to the centromere when mitosis begins We performed ChIP-Seq experiments (on mouse Trophoblast Stem cells arrested at G1; S and M stages of thecell cycle) using antibodies against histone variant H2A.Z and sequentional ChIP-re-ChIP-Seq experiments using H2A.Z antibody and H2A antibody in sequence. Combining those data sets with microarray gene expression expression data allowed us to see H2A.Z distribution over promoters of mouse coding genes in cell cycle dependant manner. Interestingly, Input also showed cell-cycle dependent effects, but histone H3 could be used as a cell-cycle independent normalisation factor. We also performed ChIP-seq with a CTCF pull-down to investigate its cell-cycle dependent relationship with heterochromatin.

ORGANISM(S): Mus musculus  

SUBMITTER: Rohan Williams   Brian Parker  Cameron Jack  Max Nekrasov 

PROVIDER: E-GEOD-41371 | ArrayExpress | 2012-11-23

SECONDARY ACCESSION(S): SRP016007GSE41371PRJNA176705

REPOSITORIES: GEO, ArrayExpress, ENA

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Publications

Histone H2A.Z inheritance during the cell cycle and its impact on promoter organization and dynamics.

Nekrasov Maxim M   Amrichova Jana J   Parker Brian J BJ   Soboleva Tatiana A TA   Jack Cameron C   Williams Rohan R   Huttley Gavin A GA   Tremethick David J DJ  

Nature structural & molecular biology 20121021 11


Although it has been clearly established that well-positioned histone H2A.Z-containing nucleosomes flank the nucleosome-depleted region (NDR) at the transcriptional start site (TSS) of active mammalian genes, how this chromatin-based information is transmitted through the cell cycle is unknown. We show here that in mouse trophoblast stem cells, the amount of histone H2A.Z at promoters decreased during S phase, coinciding with homotypic (H2A.Z-H2A.Z) nucleosomes flanking the TSS becoming heteroty  ...[more]

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