title: Chromatin restoration following nucleotide excision repair involves the incorporation of ubiquitinated H2A at damaged genomic sites.
abstract: Restoration of functionally intact chromatin structure following DNA damage processing is crucial for maintaining genetic and epigenetic information in human cells. Here, we show the UV-induced uH2A foci formation in cells lacking XPC, DDB2, CSA or CSB, but not in cells lacking XPA, XPG or XPF indicating that uH2A incorporation relied on successful damage repair occurring through either GGR or TCR sub-pathway. In contrast, XPA, XPG or XPF were not required for formation of gammaH2AX foci in asynchronous cells. Notably, the H2A ubiquitin ligase Ring1B, a component of Polycomb repressor complex 1, did not localize at DNA damage sites. However, histone chaperone CAF-1 showed distinct localization to the damage sites. Knockdown of CAF-1 p60 abolished CAF-1 as well as uH2A foci formation. CAF-1 p150 was found to associate with NER factors TFIIH, RPA p70 and PCNA in chromatin. These data demonstrate that successful NER of genomic lesions and prompt CAF-1-mediated chromatin restoration link uH2A incorporation at the sites of damage repair within chromatin.
authors.affiliation: Department of Radiology, The Ohio State University, Columbus, OH 43210, USA. zhu.49@osu.edu
authors.forename: Qianzheng
authors.initials: Q
authors.lastname: Zhu
authors.forename: Gulzar
authors.initials: G
authors.lastname: Wani
authors.forename: Hany H
authors.initials: HH
authors.lastname: Arab
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: El-Mahdy
authors.forename: Alo
authors.initials: A
authors.lastname: Ray
authors.forename: Altaf A
authors.initials: AA
authors.lastname: Wani
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism*; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism; D015894:Genome, Human / Q000235:genetics*; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D063150:Polycomb Repressive Complex 1; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D051716:Replication Protein A / Q000378:metabolism; D014157:Transcription Factors; D014158:Transcription, Genetic; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D054875:Ubiquitination; D014452:Ubiquitins / Q000378:metabolism
chemical_list: C533593:CHAF1B protein, human; C489780:CNOT8 protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D056504:Chromatin Assembly Factor-1; D002868:Chromosomal Proteins, Non-Histone; C117669:DDB2 protein, human; D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; C116662:PRC1 protein, human; D018809:Proliferating Cell Nuclear Antigen; D051716:Replication Protein A; D014157:Transcription Factors; D014452:Ubiquitins; D063150:Polycomb Repressive Complex 1; C489841:RNF2 protein, human; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex