Structural basis of the ubiquitination dependent modulation of XPC DNA repair protein.

Research Project

Project/Area Number	20770082
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Single-year Grants
Research Field	Structural biochemistry
Research Institution	Kyoto University
Principal Investigator	MAITA Ayako Kyoto University, 大学院・ヘルスバイオサイエンス研究科, 助教 (60415106)
Project Period (FY)	2008 – 2009
Project Status	Completed (Fiscal Year 2009)
Budget Amount *help	¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000) Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000) Fiscal Year 2008: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	分子認識及び相互作用 / DNA損傷 / ユビキチン修飾 / 立体構造 / NMR / X線結晶解析
Research Abstract	XPC protein specifically recognizes a certain secondary structure of the damaged DNA during the NER (Nucleotide excision repair). The ubiquitin ligase (E3) complex (cullin 4A, DDB1, DDB2 and Roc1) stimulated its activity by UV damage catalyzes ubiquitination of XPC. The ubiquitin modification of XPC promotes the DNA damage recognition by itself. However, the role of the ubiquitin modification of XPC in the NER pathway is not clear. In this study, I have attempted to elucidate the regulation of DNA damage recognition through ubiquitination of XPC using biochemical and structural analysis. Biochemical approaches was taken to identify main region of ubiquitination in XPC. On the other hand, I have tried to prepare expression constructs of several regions of XPC for the purpose of structural analysis. However, I did not succeed in preparing expression constructs for structural analysis of XPC because of low level of protein expression and protein precipitation during purification.