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G9a, ZNF644 and WIZ ChIP-seq results

ABSTRACT: The G9a mediates mono- and dimethylation of Lys9 of histone H3 at specific gene loci, which is associated with transcriptional repression. ZNF644 and WIZ contain multiple zinc finger motifs that recognize consensus DNA sequences.

ORGANISM(S): Homo sapiens

PROVIDER: GSE62616 | GEO | 2015/03/20

SECONDARY ACCESSION(S): PRJNA264545

REPOSITORIES: GEO

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A role for Widely Interspaced Zinc finger (WIZ) in retention of the G9a methyltransferase on chromatin

Project description:G9a and GLP lysine methyltransferases form a heterodimeric complex that is responsible for the bulk of cellular mono- and di-methylation on histone H3 lysine 9 (H3K9me1/me2). Widely Interspaced Zinc finger (WIZ) associates with the G9a/GLP protein complex, but its role in lysine methylation is poorly defined. Here, we show that WIZ regulates global H3K9me2 levels through a mechanism that involves retention of G9a on chromatin. We also show that WIZ-mediated chromatin loss of G9a/GLP results in altered gene expression and protein-protein interactions that are distinguishable from that of using a molecule-induced enzymatic inhibitor towards G9a/GLP – thus providing evidence that the G9a/GLP/WIZ complex has unique functions when bound to chromatin that are independent of the H3K9me2 mark DMSO, UNC0638, NS, and siWIZ treatments to HEK293T cells, assessing G9a and H3K9me2 localization, each performed in duplicate, plus one input control for each cell condition. 30 samples total.

2015-09-03 | E-GEOD-67317 | biostudies-arrayexpress

A role for Widely Interspaced Zinc finger (WIZ) in retention of the G9a methyltransferase on chromatin

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HEK293T cells treated with a G9a small molecule inhibitor (UNC0638) or vehicle (DMSO), WIZ siRNA, G9a siRNA, or scrambled (control) siRNA

Project description:Transcriptional profiling of HEK293T cells following treatment with a small molecule inhibitor of catalytic activity of the G9a lysine methyltransferase (UNC0638, 1 micromolar, 48 hours), or siRNA against WIZ or G9a (72 hours treatment).

2015-07-15 | GSE70914 | GEO

Unexpected distinct roles of the related histone H3 lysine 9 methyltransferases G9a and G9a-Like Protein in myoblasts

Project description:Lysine methyltransferases G9a and GLP (G9a- Like Protein) form functional heterodimeric complexes that establish mono- and dimethylation on histone H3 lysine 9 (H3K9me1, H3K9me2) in euchromatin. Here we describe unexpected opposite individual roles for G9a and GLP during skeletal muscle terminal differentiation. Indeed, gain- or loss-of-function assays in myoblasts showed that in consistency with previous reports that G9a inhibits terminal differentiation. But GLP plays a more complicated role in muscle differentiation since both knockdown and overexpression inhibits terminal differentiation. Unexpectedly, in contrast to G9a, we show that GLP overexpression promotes abnormal expression of muscle differentiationspecific genes in proliferating myoblasts. Transcriptomic analysis indicates that G9a and GLP regulate different sets of genes. GLP, but not G9a, inhibits at the transcriptional level proteasome subunit-encoding genes resulting in an inhibition of the proteasome activities. Subsequently, GLP, but not G9a, overexpression stabilizes MyoD that is likely to be responsible for muscle markers expression in proliferating myoblasts.

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Dynamic phosphorylation of G9a by Plk1 and PP2A modulates its catalytic activity and chromosome accessibility during mitosis

Project description:Phosphorylation of Ser10 of histone H3 (H3S10p), together with the adjacent dimethylation of Lys9 (H3K9me2), has been proposed to function as a ‘phospho-methyl switch’ to regulate mitotic chromosome dynamics and control peripheral heterochromatin localization (1, 2). Despite of immensely understanding the regulation of H3S10 phosphorylation, how this modification temporarily masks the H3K9me2 mark during mitosis is poorly understood. Herein, we report that the master mitotic kinase Plk1 phosphorylates G9a at Thr1045 (pT1045) in vitro and in vivo at early mitosis. T1045p inhibits G9a catalytic activity, leading to decrease H3K9me2 levels. RNA-sequencing analysis showed that the phospho-mimic T1045 mutant of G9a (T1045E) compromised its repressive activity on several targeted genes, which was confirmed by luciferase reporter assays. Using genome-wide epigenomic technologies, we found that T1045E mutant displayed reduced H3K9me2 level, accompanied with increased chromatin accessibility, compared to the wild-type G9a cells. Finally, we found that T1045 phosphorylation could be removed by the phosphatase PP2A at late mitosis, which inversely reactivated G9a activity on H3K9me2 level, correlated with lower levels of H3S10p. Therefore, our result may provide a mechanistic explanation for ‘phospho-methyl switch’ and highlight the importance of Plk1 and PP2A-mediated dynamic regulation of G9a during mitosis.

2023-07-17 | GSE223359 | GEO

A WIZ/Cohesin/CTCF complex anchors DNA loops to define gene expression and cell identity [RNA-Seq]

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A WIZ/Cohesin/CTCF complex anchors DNA loops to define gene expression and cell identity [Hi-C]

Project description:Purpose: Chromatin architecture is an important regulator of gene expression, but details of how DNA loops form and function remain unclear. The purpose of this study is to assess the role of Widely Interspaced Zinc-fingers protein (WIZ) in gene regulation and DNA looping. Method: Genome wide chromatin looping patterns in the presence or absence of WIZ were assessed via Hi-C in both wildtype and CRISPR-Cas9 genome edited WIZ deletion mouse embryonic stem cells (mESCs).

2020-04-14 | GSE137274 | GEO

A WIZ/Cohesin/CTCF complex anchors DNA loops to define gene expression and cell identity [ChIP-Seq]

Project description:Purpose: Chromatin architecture is an important regulator of gene expression, but details of how DNA loops form and function remain unclear. The purpose of this study is to assess the role of Widely Interspaced Zinc-fingers protein (WIZ) in gene regulation and DNA looping. Method: Genome wide binding patterns of known architectural proteins in the presence or absence of WIZ were assessed via chromatin immunoprecipitation followed by high throughput sequencing (ChIP-seq) in both wildtype and CRISPR-Cas9 genome edited WIZ deletion mouse embryonic stem cells (mESCs).

2020-04-14 | GSE137272 | GEO

Genomic sensitization in response to G9a repression following repeated cocaine exposure

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G9a regulates temporal preimplantation developmental program and lineage segregation in blastocyst

Project description:Early mouse development is regulated and accompanied by dynamic changes in chromatin modifications, including G9a-mediated histone H3 lysine 9 dimethylation (H3K9me2). Previously, we provided insights into its role in post-implantation development (Zylicz et al., 2015). Here we explore the impact of depleting the maternally inherited G9a in oocytes on development shortly after fertilisation. We show that H3K9me2 normally accumulates at 8- cell stage to promote timely repression of a subset of 4-cell stage specific genes. Loss of maternal inheritance of G9a disrupts the gene regulatory network resulting in developmental delay and destabilisation of inner cell mass lineages by the late blastocyst stage. Our results indicate a vital role of this maternally inherited epigenetic regulator in creating conductive conditions for developmental progression and on cell fate choices.

2018-05-15 | GSE106790 | GEO

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