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Molecular and Cellular Biology, July 2005, p. 5664-5674, Vol. 25, No. 13
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.13.5664-5674.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Centrin 2 Stimulates Nucleotide Excision Repair by Interacting with Xeroderma Pigmentosum Group C Protein

Ryotaro Nishi,1,2,3,4 Yuki Okuda,1,3 Eriko Watanabe,1 Toshio Mori,5 Shigenori Iwai,6 Chikahide Masutani,2,4 Kaoru Sugasawa,1,2* and Fumio Hanaoka1,2,4

Cellular Physiology Laboratory, RIKEN Discovery Research Institute,1 SORST, Japan Science and Technology Agency, Saitama,2 Graduate School of Pharmaceutical Sciences,3 Graduate School of Frontier Biosciences, Osaka University, Osaka,4 Radioisotope Center, Nara Medical University, Nara,5 Graduate School of Engineering Science, Osaka University, Osaka, Japan6

Received 30 January 2005/ Returned for modification 2 March 2005/ Accepted 5 April 2005

Xeroderma pigmentosum group C (XPC) protein plays a key role in DNA damage recognition in global genome nucleotide excision repair (NER). The protein forms in vivo a heterotrimeric complex involving one of the two human homologs of Saccharomyces cerevisiae Rad23p and centrin 2, a centrosomal protein. Because centrin 2 is dispensable for the cell-free NER reaction, its role in NER has been unclear. Binding experiments with a series of truncated XPC proteins allowed the centrin 2 binding domain to be mapped to a presumed {alpha}-helical region near the C terminus, and three amino acid substitutions in this domain abrogated interaction with centrin 2. Human cell lines stably expressing the mutant XPC protein exhibited a significant reduction in global genome NER activity. Furthermore, centrin 2 enhanced the cell-free NER dual incision and damaged DNA binding activities of XPC, which likely require physical interaction between XPC and centrin 2. These results reveal a novel vital function for centrin 2 in NER, the potentiation of damage recognition by XPC.

* Corresponding author. Mailing address: Cellular Physiology Laboratory, RIKEN Discovery Research Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. Phone: 81 48 467 9532. Fax: 81 48 462 4673. E-mail: sugasawa{at}riken.jp.

Molecular and Cellular Biology, July 2005, p. 5664-5674, Vol. 25, No. 13
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.13.5664-5674.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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