TY - JOUR
T1 - Sequential assembly of the nucleotide excision repair factors in vivo
AU - Volker, Marcel
AU - Moné, Martijn J.
AU - Karmakar, Parimal
AU - Van Hoffen, Anneke
AU - Schul, Wouter
AU - Vermeulen, Wim
AU - Hoeijmakers, Jan H.J.
AU - Van Driel, Roel
AU - Van Zeeland, Albert A.
AU - Mullenders, Leon H.F.
N1 - Funding Information:
The authors would like to thank Drs. Harry Vrieling and Niels de Wind for critical reading of the manuscript. This work was supported by the Netherlands Organization for Scientific Research (ALW) program “Nucleotide Excision Repair in Its Nuclear Context” and by European Union project number QRLT-1999-00181.
PY - 2001
Y1 - 2001
N2 - Here, we describe the assembly of the nucleotide excision repair (NER) complex in normal and repair-deficient (xeroderma pigmentosum) human cells, employing a novel technique of local UV irradiation combined with fluorescent antibody labeling. The damage recognition complex XPC-hHR23B appears to be essential for the recruitment of all subsequent NER factors in the preincision complex, including transcription repair factor TFIIH. XPA associates relatively late, is required for anchoring of ERCC1-XPF, and may be essential for activation of the endonuclease activity of XPG. These findings identify XPC as the earliest known NER factor in the reaction mechanism, give insight into the order of subsequent NER components, provide evidence for a dual role of XPA, and support a concept of sequential assembly of repair proteins at the site of the damage rather than a preassembled repairosome.
AB - Here, we describe the assembly of the nucleotide excision repair (NER) complex in normal and repair-deficient (xeroderma pigmentosum) human cells, employing a novel technique of local UV irradiation combined with fluorescent antibody labeling. The damage recognition complex XPC-hHR23B appears to be essential for the recruitment of all subsequent NER factors in the preincision complex, including transcription repair factor TFIIH. XPA associates relatively late, is required for anchoring of ERCC1-XPF, and may be essential for activation of the endonuclease activity of XPG. These findings identify XPC as the earliest known NER factor in the reaction mechanism, give insight into the order of subsequent NER components, provide evidence for a dual role of XPA, and support a concept of sequential assembly of repair proteins at the site of the damage rather than a preassembled repairosome.
UR - http://www.scopus.com/inward/record.url?scp=17944361949&partnerID=8YFLogxK
U2 - 10.1016/S1097-2765(01)00281-7
DO - 10.1016/S1097-2765(01)00281-7
M3 - Article
C2 - 11511374
AN - SCOPUS:17944361949
SN - 1097-2765
VL - 8
SP - 213
EP - 224
JO - Molecular Cell
JF - Molecular Cell
IS - 1
ER -