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The HP1 protein interactome of mouse embryonic stem cells

ABSTRACT: The family of Heterochromatin Protein 1 (HP1) consists of highly conserved proteins, which have important functions in the nucleus of eukaryotic cells. In mammals there are three HP1 paralogs: HP1(alpha), Hp1(beta), and Hp1(gamma)They are encoded by the Cbx5, Cbx1, and Cbx3 genes, respectively. Hp1 and Hp1 stably interact with Chd4 and Adnp to form the ChAHP complex. In this project, Chd4, Adnp, and the three Cbx genes were endogenously tagged with a FLAG-Avi tag in mouse embryonic stem cells. The tagged proteins were subjected to tandem-affinity purification and analysis by mass spectrometry.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryonic Stem Cell

SUBMITTER: Vytautas Iesmantavicius

LAB HEAD: Marc Bühler

PROVIDER: PXD006226 | Pride | 2018-05-22

REPOSITORIES: Pride

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Activity-dependent neuroprotective protein recruits HP1 and CHD4 to control lineage-specifying genes.

Ostapcuk Veronika V Mohn Fabio F Carl Sarah H SH Basters Anja A Hess Daniel D Iesmantavicius Vytautas V Lampersberger Lisa L Flemr Matyas M Pandey Aparna A Thomä Nicolas H NH Betschinger Joerg J Bühler Marc M

Nature 20180523 7707

De novo mutations in ADNP, which encodes activity-dependent neuroprotective protein (ADNP), have recently been found to underlie Helsmoortel-Van der Aa syndrome, a complex neurological developmental disorder that also affects several other organ functions <sup>1</sup> . ADNP is a putative transcription factor that is essential for embryonic development <sup>2</sup> . However, its precise roles in transcriptional regulation and development are not understood. Here we show that ADNP interacts with ...[more]

PMID: 29795351

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Compartmentalization of HP1 proteins in pluripotency acquisition and maintenance

Project description:Embryonic stem cells (ESCs) have a hyperdynamic chromatin structure characterized by fewer discrete foci of heterochromatin protein 1 family of proteins (HP1) compared to somatic cells. During reprogramming of somatic cells, depletion of HP1γ early, reduced, while depletion later, enhanced the generation of induced pluripotent stem cells (iPSCs); concomitant with a change from a centromeric to nucleoplasmic localization. To identify the interactome of HP1γ in different biochemical environment in ESCs, we compared protein complexes of HP1γ at 0.42M salt (Dignam et al, 1983), with micrococcal nuclease (MCN) digestion with 0.3M NaCl, and MCN with 0.5M NaCl. To understand the effectors of the change in localization, we isolated protein complexes containing HP1γ in ESCs and compared the profile to that in partially reprogrammed intermediates called pre-iPSCs. Given the differential distribution of the HP1 family in ESCs, we also compared the protein interactome of HP1γ in ESCs with that of HP1α and HP1β. In addition, we probed histone associations of HP1γ in ESCs further by querying the histone post-translational modifications that were detectable. The HP1 proteins themselves are decorated with multiple PTMs, several of which we have found to be novel. Taken together our results reveal the complex contribution of the HP1 proteins to pluripotency.

2017-12-13 | PXD006817 | Pride

GFI1 tethers the NuRD complex to open and transcriptionally active chromatin in myeloid progenitors

Project description:GFI is a DNA binding transcriptional repressor that regulates myeloid differentiation. Here, we show that GFI1 interacts with the chromodomain helicase CHD4 and other components of the nucleosome remodeling and deacetylase (NuRD) complex. Our data demonstrated that GFI1 and GFI1/CHD4 complexes occupy sites of open chromatin enriched for histone marks associated with active transcription or different sets of genes that are either enriched for IRF1 or SPI-1 consensus binding sites. In addition, our study provided evidence that GFI1 affects the chromatin remodeling activity of the NuRD complex. Overall, our results indicate that GFI1/CHD4 complexes control chromatin openness and histone modifications differentially to regulate target genes, which govern the immune response, nucleosome organization, or metabolic processes.

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CBX3 Regulates Efficient RNA Processing Genome Wide

Project description:HP1 alpha, beta, and gamma play an evolutionarily conserved role in the formation and maintenance of heterochromatin. In addition, some HP1 family members may also participate in transcriptional regulation of genes. Recently, HP1 gamma binding to the bodies of a subset of genes has been observed in human and murine cells. However, the generality of this phenomenon and the role HP1 gamma may play in this context are unknown. Genome-wide localization analysis reveals broad and general HP1 gamma binding at actively transcribed genes, which strongly correlates with gene activity across multiple cell types. Unexpectedly, we find HP1 gamma binding is necessary for efficient RNA processing of its target genes. Loss of HP1 gamma results in defective recruitment of splicing factors including U1-70K, and leads to accumulation of unspliced nascent transcripts genome-wide. Therefore, our data argue that the primary role of HP1 gamma is to ensure efficient RNA processing in mammalian cells. Genome-wide localization and function of HP1 gamma in multiple cell types using ChIP-chip, ChIP-seq, and RNA-seq

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Multiple Overlapping Mammalian Chromatin Remodeling Systems Collaborate Genome-wide at Dynamic Chromatin Transitions (ChIP-Seq)

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2013-12-24 | E-GEOD-53583 | biostudies-arrayexpress

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2013-12-24 | E-GEOD-53584 | biostudies-arrayexpress

Proteo-Genomic Characterization and Mapping of Nucleosomes Decoded by Brd and HP1 Proteins

Project description:This SuperSeries is composed of the following subset Series: GSE39579: Proteo-Genomic Characterization and Mapping of Nucleosomes Decoded by Brd and HP1 Proteins (Chip-Seq data) GSE39580: Proteo-Genomic Characterization and Mapping of Nucleosomes Decoded by Brd and HP1 Proteins (expression data) Refer to individual Series

2012-07-30 | E-GEOD-39581 | biostudies-arrayexpress

Defining the interactome of CHD4 in fusion-positive rhabdomyosarcoma

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2020-08-11 | PXD015231 | Pride

Mus musculus

Project description:ChAHP (Adnp, Chd4, HP1) mediates sequence-specific recruitment of HP1 independent of H3K9 methylation

| PRJNA383413 | ENA

ChIP-seq of NuRD components and pluripotency factors in wild-type, Mbd3 and Sall4/1 mutant embryonic stem cells

Project description:Members of the Nucleosome Remodeling and Deacetylase (NuRD) complex Mbd3 and Mi2beta (Chd4) along with pluripotency regulators Nanog, Oct4, Klf4 and Esrrb were profiled for chromatin association by ChIP-seq in mouse embryonic stem cells mutant for Mbd3, Sall4 and/or the related Sall1 pluripotency-associated factors.

2016-08-09 | E-MTAB-4565 | biostudies-arrayexpress

ChAHP (Adnp, Chd4, HP1) mediates sequence-specific recruitment of HP1 independent of H3K9 methylation

Project description:Activity-dependent neuroprotective protein (ADNP) is one of the most frequent autism spectrum disorder-associated gene products known to date. Here we show that Adnp interacts with the chromatin remodeler Chd4 and the heterochromatin protein HP1 to form a stable complex, which we refer to as ChAHP. Genetic ablation of ChAHP components or DNA binding sites in embryonic stem cells prematurely activates lineage-specific genes, revealing an important role for Adnp in restraining the differentiation capacity of pluripotent cells. Adnp targets the ChAHP complex to specific sequence motifs at euchromatic loci, representing an H3K9 methylation-independent mechanism of HP1 recruitment and gene silencing.

2018-05-23 | GSE97945 | GEO

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