TY - JOUR
T1 - Chromatin restoration following nucleotide excision repair involves the incorporation of ubiquitinated H2A at damaged genomic sites
AU - Zhu, Qianzheng
AU - Wani, Gulzar
AU - Arab, Hany H.
AU - El-Mahdy, Mohamed A.
AU - Ray, Alo
AU - Wani, Altaf A.
N1 - Funding Information:
The work carried out by Q.Z., G.W., M.E., A.R. and A.A.W. was supported by Public Health Service Grants ES2388 and ES12991 from National Institute of Environmental Health Sciences and CA93413 from National Cancer Institute. We thank Haroon Haque for excellent technical assistance and Qi-en Wang, The Ohio State University, for advice on immunofluorescence experimentation. We are grateful to Yoshihiro Nakatani, Harvard Medical School, for providing HeLa-DDB2 cells.
PY - 2009/2/1
Y1 - 2009/2/1
N2 - 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 γH2AX 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.
AB - 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 γH2AX 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.
KW - CAF-1
KW - Chromatin assembly
KW - DNA damage
KW - DNA repair
KW - Nucleotide excision repair
KW - Ubiquitinated histone H2A
UR - https://www.scopus.com/pages/publications/58149159543
U2 - 10.1016/j.dnarep.2008.11.007
DO - 10.1016/j.dnarep.2008.11.007
M3 - Article
C2 - 19059499
AN - SCOPUS:58149159543
SN - 1568-7864
VL - 8
SP - 262
EP - 273
JO - DNA Repair
JF - DNA Repair
IS - 2
ER -