Dataset Information

ANP32e binds histone H2A.Z in a cell cycle-dependent manner and regulates its protein stability in the cytoplasm

ABSTRACT: ANP32e, a chaperone of H2A.Z, is receiving increasing attention because of its link to cancer growth and progression. An unanswered question is whether ANP32e regulates H2A.Z dynamics during the cell cycle; if so, this could have clear implications for the proliferation of cancer cells. Using the human U2OS cancer cell line model system, we have confirmed that ANP32e regulates the growth of these cells. ANP32e preferentially interacts with H2A.Z during G1 phase of the cell cycle. Unexpectedly, however, ANP32e does not mediate the removal of H2A.Z from chromatin, is not a stable component of the p400 remodeling complex, and is not strongly associated with chromatin. Instead, most ANP32e is in the cytoplasm. Here, ANP32e preferentially interacts with H2A.Z in G1 phase in response to an increase in H2A.Z protein abundance and regulates its protein stability. This G1-specific interaction between ANP32e and H2A.Z is also observed in the nucleoplasm but is unrelated to any change in H2A.Z abundance. Collectively, these results challenge the idea that ANP32e is involved in regulating the abundance of H2A.Z in chromatin as part of a chromatin remodeling complex. Rather, we propose that ANP32e acts as a molecular chaperone that maintains the soluble pool of H2A.Z by regulating its protein stability and acting as a buffer in response to cell cycle-dependent changes in H2A.Z abundance.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

SUBMITTER: Gene Hart-Smith

LAB HEAD: David J Tremethick

PROVIDER: PXD033229 | Pride | 2024-05-23

REPOSITORIES: Pride

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ANP32e Binds Histone H2A.Z in a Cell Cycle-Dependent Manner and Regulates Its Protein Stability in the Cytoplasm.

Dijkwel Yasmin Y Hart-Smith Gene G Kurscheid Sebastian S Tremethick David J DJ

Molecular and cellular biology 20240314 2

ANP32e, a chaperone of H2A.Z, is receiving increasing attention because of its association with cancer growth and progression. An unanswered question is whether ANP32e regulates H2A.Z dynamics during the cell cycle; this could have clear implications for the proliferation of cancer cells. We confirmed that ANP32e regulates the growth of human U2OS cancer cells and preferentially interacts with H2A.Z during the G1 phase of the cell cycle. Unexpectedly, ANP32e does not mediate the removal of H2A.Z ...[more]

PMID: 38482865

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Project description: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 1,2, 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 surveyed whole genome expression using microarrays, in combination with H2A.Z ChIP-Seq (Illumina) data, to study the H2A.Z distribution on promoters of coding genes in different stages of the cell cycle in Trophoblast Stem (TS) cells. These data are published in Nekrasov et al., H2AZ inheritance during the cell cycle and it's impact on promoter organization and dynamics, Nature Structural and Molecular Biology (in press: accepted for publication on 24 Sep 2012). Mouse Trophoblast stem (TS) cells were grown in cell culture medium. By adding specific drugs cells were arrested at particular stages of the cell cycle: G1; S and M. Total RNA was extracted from TS cells arrested at G1; S and M stages of cell cycle using TRizol (Invitrogen) extraction protocol and purified further the by RNAeasy Extraction Kit (Qiagen). RNA quality was assesed on Bioanalyzer total RNA chip. RNA samples were reverse transcribed and resulted cDNA samples were subjected to DNA microarray analysis (in triplicate) using the GeneChip Mouse Gene 1.0 ST Array (Affymetrix). Robust Multichip Average (RMA) correction and probe set summaries were obtained using Affymetrix Power Tools software annotation (Affymetrix) based on the mm9 mouse genome. Unless otherwise stated, all subsequent expression analyses (see Supplementary Methods) were performed in the R statistical computing environment (version 2.11.1).

			Action	DRS
	7_nuc_WT_G1.raw	Raw
	8_nuc_WT_M.raw	Raw
	YD17_nuc_ANP32e_G1_1.raw	Raw
	YD18_nuc_ANP32e_G1_2.raw	Raw
	YD19_nuc_ANP32e_M1.raw	Raw