Dataset Information

Searching for interactors of endogenous SLX4

ABSTRACT: The project was aimed at identifying binding partners of endogenous SLX4 in HeLa cells. Whole cell extracts from HeLa Flp-In T-Rex cells were used for immunoprecipitation performed with anti-SLX4 antibodies from Bethyl Laboratories (A302-270A and A302-269A), either used alone or in combination. To discriminate bona fide SLX4 partners from non-specific proteins recognized by the anti-SLX4 antibodies, cells were transfected with control (Luc) siRNA or a siRNA targeting SLX4. After immunoprecipitation, protein complexes were analysed by LC-MSMS.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

SUBMITTER: AUDEBERT Stephane

LAB HEAD: Pierre-Henri Gaillard

PROVIDER: PXD029084 | Pride | 2022-02-16

REPOSITORIES: Pride

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Publications

SLX4 dampens MutSα-dependent mismatch repair.

Guervilly Jean-Hugues JH Blin Marion M Laureti Luisa L Baudelet Emilie E Audebert Stéphane S Gaillard Pierre-Henri PH

Nucleic acids research 20220301 5

The tumour suppressor SLX4 plays multiple roles in the maintenance of genome stability, acting as a scaffold for structure-specific endonucleases and other DNA repair proteins. It directly interacts with the mismatch repair (MMR) protein MSH2 but the significance of this interaction remained unknown until recent findings showing that MutSβ (MSH2-MSH3) stimulates in vitro the SLX4-dependent Holliday junction resolvase activity. Here, we characterize the mode of interaction between SLX4 and MSH2, ...[more]

PMID: 35166826

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Project description:Analysis of transcriptional regulation in human cells has implicated a large number of promoter-specific DNA-binding proteins that regulate transcription via diverse mechanisms. In some cases, these DNA-sequence-specific factors associate with intermediaries that orchestrate interactions with the chromatin-modifying enzymes. One such intermediary is HCF-1 (host-cell factor-1; HCFC1). HCF-1, first identified for its involvement in herpes-simplex virus transcription and subsequently shown to be an important cell-cycle regulator, has a poorly defined role in genome-wide transcriptional regulation. We show here, by chromatin immunoprecipitation followed by high-throughput sequence analysis (ChIP-seq), that HCF-1 is a major transcriptional start site associated factor, whose promoter association correlates positively with transcriptional activity. Thus, in HeLa cells HCF-1 is observed on 5400 generally active CpG-island promoters. Examination of the DNA sequences underlying the HCF-1-binding sites revealed three sequence motifs associated with the binding of (i) ZNF143 and Ronin/THAP11, (ii) GABP, and (iii) YY1 sequence-specific transcription factors. Subsequent ChIP-seq analysis of these four transcription factors revealed a large co-association of HCF-1 with these four transcription factors at approximately 90% of HCF-1-bound promoters. These studies suggest that a relatively small number of transcription factors -- some (ZNF143 and Ronin/THAP11) in novel combinations -- play a major role in HeLa-cell transcriptional regulation in association with HCF-1. This experiment includes the sequencing data of material obtained from chromatin immunoprecipitation using antibodies against HCFC1(antibodies against the C-subunit and the N-subunit), PolII(antibody against the RPB2 subunit), H3K36ME3 and H3K4Me3 histone modifications, ZNF143, THAP11, GABPalpha and YY1. It also includes control data of the Input material. All of them were done using HeLa cycling cells.

Dataset Information

Searching for interactors of endogenous SLX4

Publications

SLX4 dampens MutSα-dependent mismatch repair.

Similar Datasets

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