Changes in Chromatin Structure and Its Regulation in DNA Repair

• Project/Area Number: 16209003
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Osaka University
• Principal Investigator: HANAOKA Humio Osaka University, Graduate School of Frontier Biosciences, Professor (50012670)
• Co-Investigator(Kenkyū-buntansha): MASUTANI Chikahide Osaka University, Graduate School of Frontier Biosciences, Associate Professor (40241252)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥47,970,000 (Direct Cost: ¥36,900,000、Indirect Cost: ¥11,070,000); Fiscal Year 2006: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000); Fiscal Year 2005: ¥15,990,000 (Direct Cost: ¥12,300,000、Indirect Cost: ¥3,690,000); Fiscal Year 2004: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
• Keywords: DNA repair / chromatin structure / nucleotide excision repair / XPC protein / UV-DDB / ubiquitylation / centrin 2 / condensin / ヒストンアセチル化 / CBP / NER / DDB因子

Research Abstract

In order to investigate how chromatin structure influences DNA repair mechanism such as nucleotide excision repair (NER) and translesion synthesis (TLS), we analyzed molecular mechanisms of NER and TLS with special reference to chromatin structure. Followings are summary of our results.
1) In global genome NER, two mammalian homologs of Rad23, HR23A and HR23B, were found to be equivalent in terms of NER functions. In other words, either one of them is enough for NER, if the protein level is sufficient.
2) The specific binding of the XPC complex to (6-4) photoproducts was competitively inhibited by the addition of a large excess ofundamaged naked DNA. In contrast, the addition of undamaged nucleosomal DNA as a competitor suppressed the inhibitory effect. Although nucleosomes positioned on the damaged site inhibited the binding of the XPC complex, the presence of nucleosomes in undamaged DNA regions may help specific binding of the XPC complex to damaged sites by excluding its non-specific binding to damaged DNA regions.
3) XPC undergoes reversible ubiquitylation upon UV irradiation of cells and that this depends on the presence of functional UV-DDB activity. XPC and UV-DDB were demonstrated to interact physically, and both are polyubiquitylated by the recombinant UV-DDB-ubiquitin ligase complex.
4) We have identified three amino acid residues in human XPC protein that are essential for the binding to centrin 2. Alanine substitutions of these amino acids in XPC protein abrogated interaction with centrin 2. Human cell lines stably expressing the mutant XPC protein exhibited a significant reduction on global genome NER activity. Centrin 2 enhanced the cell-free NER dual incision and damaged DNA binding activities of XPC.
5) CK2 phosphorylates condensin I during interphase and reduced its supercoiling activity. On the other hand, CK2-dependent phosphorylation decreases on chromosomes during mitosis. Thus condensin I is regulated by CK2 phosphorylation.

Research Products

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  • Journal Title: EMBO J. 25(22) Pages: 5339-5348
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Conserved XPB core structure and motifs for DNA unwinding :implications for pathway selection of transcription or excision repair (2006)
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  • Journal Title: Mol.Cell 22(1) Pages: 27-37
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Negative regulation of condensin I by CK2-mediated phosphorylation. (2006)
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  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Conserved XPB core structure and motifs for DNI unwinding : implications for pathway selection o transcription or excision repair. (2006)
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  • Journal Title: Mol. Cell 22(1) Pages: 27-37
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• [Journal Article] Negative regulation of condensin I by CK2-mediated phosphorylation. (2006)
  • Author(s): Takemoto, A. et al.
  • Journal Title: EMBO J. 25(22) Pages: 5339-5348
• [Journal Article] Crystal structure of SUMO-3-modified thymine-DNA glycosylase. (2006)
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• [Journal Article] Conserved XPB core structure and motifs for DNA unwinding : implications for pathway selection of transcription or excision repair. (2006)
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  • Journal Title: Mol. Cell 22(1) Pages: 27-37
• [Journal Article] Centrin 2 stimulates nucleotide excision repair by interacting with xeroderma pigmentosum group C protein (2005)
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  • Journal Title: Mol.Cell.Biol. 25(13) Pages: 5664-5674
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] UV-induced ubiquitylayion of XPC protein mediated by UV-DDB-ubiquitin ligase complex (2005)
  • Author(s): Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Chu, G., Mori, T., Iwai, S., Tanaka, K., Tanaka, K., and Hanaoka, F.
  • Journal Title: Cell 121(3) Pages: 387-400
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Nucleosomal structure of undamaged DNA regions suppresses the non-specific DNA binding of the XPC complex (2005)
  • Author(s): Ysuda, T., Sugasawa, K., Shimizu, Y., Iwai, S., Shiomi, T., and Hanaoka, F.
  • Journal Title: DNA Repair 4(3) Pages: 389-395
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Centrin 2 stimulates nucleotide excision repair by interacting with xeroderma pigmentosum group C protein. (2005)
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  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex. (2005)
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  • Journal Title: Cell 121(3) Pages: 387-400
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nucleosomal structure of undamaged DNA regions suppresses the non-specific DNA binding of the XPC complex. (2005)
  • Author(s): Ysuda, T., Sugasawa, K., Shimizu, Y., Iwai, S., Shiomi, T., Hanaoka, F.
  • Journal Title: DNA Repair 4(3) Pages: 389-395
  • Description: 「研究成果報告書概要(欧文)」より
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• [Journal Article] Centrin 2 stimulates nucleotide excision repair by interacting with xeroderma pigmentosum group C protein. (2005)
  • Author(s): Nishi, R. et al.
  • Journal Title: Mol.Cell.Biol. 25 Pages: 5664-5674
• [Journal Article] Crystal Structure of SUMO1-conjugated thymine DNA glycosylase. (2005)
  • Author(s): Baba, D. et al.
  • Journal Title: Nature 435 Pages: 979-982
• [Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex. (2005)
  • Author(s): Sugasawa, K.. et al.
  • Journal Title: Cell 121 Pages: 387-400
• [Journal Article] UV-induced ubiquitylation of XPC protein modiated by UV-DDB-ubiquitin ligase complex. (2005)
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  • Journal Title: Cell (in press)
• [Journal Article] Nucleosomal structure of undamaged DNA regions suppresses the non-specific DNA binding of the XPC complex. (2005)
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  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Relative levels of the two mammalian Rad23 homologs determine composition and stability of the xeroderma pigmentosum group C protein complex. (2004)
  • Author(s): Okuda, Y., Nishi, R., Ng, J. M. Y., Vermeulen, W., van der Horst, G. T. J., Mori, T., Hoeijmakers, J. H. J., Hanaoka, E. Sugasawa, K.
  • Journal Title: DNA Repair 3(10) Pages: 1285-1295
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• [Journal Article] Relative levels of the two mammalian Rad23 homologs determine composition and stability of the xeroderma pigmentosum group C protein complex. (2004)
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• [Book] がん研究のいま(1)発がんの分子機構と防御 (2006)
  • Author(s): 笹月 健彦, ら
  • Total Pages: 197
  • Publisher: 東京大学出版会
  • Description: 「研究成果報告書概要(和文)」より
• [Book] がん研究のいま(1)発がんの分子機構と防御 (2006)
  • Author(s): 笹月 健彦ら
  • Total Pages: 197
  • Publisher: 東京大学出版会