title: Acetyltransferase p300 collaborates with chromodomain helicase DNA-binding protein 4 (CHD4) to facilitate DNA double-strand break repair.
abstract: Chromatin remodelling is critical for repairing DNA damage and maintaining genomic integrity. Previous studies have reported that histone acetyltransferase p300 and ATP-dependent chromatin remodeler chromodomain helicase DNA-binding protein 4 (CHD4) functions, respectively, in DNA double-strand breaks (DSBs) repair. But the physiological significance of their interaction remains elusive. Here, we showed that p300 and CHD4 were both recruited to the sites of DSBs. Their ablation led to impaired DSBs repair and sensitised cells to laser and the anti-cancer drug, etoposide. Using DR-GFP and EJ5-GFP reporter systems, we found that knockdown of p300 or CHD4 impaired the homologous recombination (HR) repair but no the non-homologous end joining (NHEJ) repair. Furthermore, p300 or CHD4 knockdown respectively suppressed the recruitment of replication protein A (RPA), a key protein for HR, to the DSB sites. In addition, immunofluorescence results showed that knockdown of p300 reduced the recruitment of CHD4 at DSB sites. In turn, CHD4 knockdown also decreased p300 assembly. Moreover, immunoprecipitation and purified protein pull down assay revealed that p300 physically interacted with CHD4 at DNA damage sites, and this interaction was dependent on the chromodomain and ATPase/helicase domain of CHD4 and the CH2, Bd and HAT domains of p300. These results indicate that p300 and CHD4 could function cooperatively at DSB sites and provide a new insight into the detailed crosstalk among the chromatin remodelling proteins.
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and Department of Bioscience, Changchun Normal University, Changchun, Jilin, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Qi
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Chen
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and.
authors.forename: Ting
authors.initials: T
authors.lastname: Xiao
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and.
authors.forename: Ruoxi
authors.initials: R
authors.lastname: Wang
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and.
authors.forename: Ting
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Bioscience, Changchun Normal University, Changchun, Jilin, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Han
authors.affiliation: Institute of Genetics and Cytology, School of Life Sciences, Northeast Normal University, #5268, Renmin Street, Changchun, Jilin, China and zengx779@nenu.edu.cn.
authors.forename: Xianlu
authors.initials: X
authors.lastname: Zeng
mesh_terms: D001324:Autoantigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D042002:Chromatin Assembly and Disassembly; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004260:DNA Repair; D015870:Gene Expression; D059765:Homologous Recombination; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D034622:RNA Interference; D034741:RNA, Small Interfering; D050880:p300-CBP Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000099224:p300-CBP-Associated Factor
chemical_list: D001324:Autoantigens; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D034741:RNA, Small Interfering; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C115323:CHD4 protein, human