Dataset Information

EPIGENETIC MODULATION IN LIVE CELLS USING A PEPTIDE OF THE C-TERMINAL UNSTRUCTURED TAIL OF H2A.Z

ABSTRACT: H2A.Z-nucleosomes participate in both euchromatic and heterochromatic scenarios and it has proven difficult to reveal correlation of the disparate roles with the stability features imparted by H2A.Z on the nucleosomes. Using a robust in situ assay of nucleosome stability and cell lines expressing only engineered forms of the variant we show that a major fraction of H2A.Z is released from the nucleosomes of peripheral heterochromatin at unusually high salt concentration, an observation reproduced with reconstituted nucleosomes. In cells expressing H2A.Z lacking its C-terminal tail, or upon addition of the tail peptide to control nuclei, canonical stability features are restored, the peripheral heterochromatin becomes dispersed and an increase of nuclease sensitivity, mainly at lamina-associated domains, ensues. Binding of the peptide to nucleosomes was detected by fluorescence correlation spectrometry. The peptide, when introduced into live cells, also induces reorganization of chromatin and decreases MYC expression, offering means for targeted epigenetic modulation.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Uladzislau Vadadokhau

LAB HEAD: Csősz Éva

PROVIDER: PXD040998 | Pride | 2025-05-06

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20081005a_ImreL_NaCl_kezelt.raw	Raw
	20181005a_ImreL_NaCl_kezelt_Proteingroups.xlsx	Xlsx
	20210212a_SzaboG_2.raw	Raw
	20210212a_SzaboG_2_Proteingroups.xlsx	Xlsx
	20210215a_SzaboG_4.raw	Raw

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Publications

Epigenetic modulation via the C-terminal tail of H2A.Z.

Imre László L Nánási Péter P Benhamza Ibtissem I Enyedi Kata Nóra KN Mocsár Gábor G Bosire Rosevalentine R Hegedüs Éva É Niaki Erfaneh Firouzi EF Csóti Ágota Á Darula Zsuzsanna Z Csősz Éva É Póliska Szilárd S Scholtz Beáta B Mező Gábor G Bacsó Zsolt Z Timmers H T Marc HTM Kusakabe Masayuki M Balázs Margit M Vámosi György G Ausio Juan J Cheung Peter P Tóth Katalin K Tremethick David D Harata Masahiko M Szabó Gábor G

Nature communications 20241024 1

H2A.Z-nucleosomes are present in both euchromatin and heterochromatin and it has proven difficult to interpret their disparate roles in the context of their stability features. Using an in situ assay of nucleosome stability and DT40 cells expressing engineered forms of the histone variant we show that native H2A.Z, but not C-terminally truncated H2A.Z (H2A.Z∆C), is released from nucleosomes of peripheral heterochromatin at unusually high salt concentrations. H2A.Z and H3K9me3 landscapes are reor ...[more]

PMID: 39448645

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Project description:Our group is interested in epithelial-to-mesenchymal transition (EMT), in particular, TGF-beta induced EMT. TGF-beta signalling has been shown to be an important factor in the induction of EMT and it has been demonstrated that adding TGF-beta to epithelial cells in culture is a convenient way to study the process of EMT. In response to TGF-beta, Smad2 and 3 are activated, and form complexes with Smad4, which then regulate transcription of target genes through interactions with other DNA binding transcription factors. In the induction of EMT, the activated Smads mediate transcriptional regulation through three families of transcription factors, resulting in repression of epithelial marker gene expression and activation of mesenchymal gene expression (Xu J, et al. 2009) Also investigated in this study is the role of H2A.Z in EMT. H2A.Z is an evolutionary conserved and a metazoan essential histone variant of the H2A class. Mice deficient in H2A.Z die during early development but the reason for this is unknown (Faast et al. 2001). Previously, our laboratory showed that the loss of H2A.Z in Xenpous laevis impaired cell movement required for the formation of the mesoderm and neural crest (Ridgway et al. 2004). Given that mesoderm formation is critically dependent upon EMT, we therefore wondered whether H2A.Z might be a chromatin regulator of EMT. We transfected MDCK cells with a lentiviral vector to express a construct encoding an shRNA targeting canine H2A.Z as we wanted to test the hypothesis that H2A.Z is involved in the maintenance of cellular identity and that its loss might trigger de-differentiation. In order to investigate changes in histone variant H2A.Z occupancy associated with TGF-beta induced epithelial-to-mesenchymal transition (EMT) we performed H2A.Z ChIP-Seq in untreated and TGFb-treated MDCK cells. The MDCK cell line has been extensively used as a model system for EMT because they convert fully from the epithelial to the mesenchymal state in response to TGF-beta. Please note that RNA-seq data generated in conjunction to this ChIP-seq data set were also deposited at ArrayExpress under accession number E-MTAB-5628 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-5628 ).