2007 Fiscal Year Final Research Report Summary
Repair mechanism of UV-induced DNA damage and roles of ubiquitination
| Project/Area Number |
17310040
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Risk sciences of radiation/Chemicals
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| Research Institution | Kobe University (2007)
The Institute of Physical and Chemical Research (2005-2006) |
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Principal Investigator |
SUGASAWA Kaoru Kobe University, Organization of Advanced Science ans Technology Biosignal Research Center, Professor (70202124)
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| Project Period (FY) |
2005 – 2007
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| Keywords | nucleotide excision repair / xeroderma nigmentosum / DNA damage recognition / ubinuitination / XPC / UV-DDB |
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| Research Abstract |
Biochemical analyses were carried out on ubiquitylation of XPC and DDB2 by the UVDDB-ubiquitin hose (E3) complex induced in UV-irradiated cells. Upon ubiquitylation of DDB2, UVDDB lost its strong binding activity to UV-damaged DNA, whereas ubiquitylated XPC still retained its DNA binding activity. Furthermore, elongation of polyubiquitin chains seemed to be required for in vitro nucleotide excision repair (NER) reactions only when UV-DDB was present. Based on these results, we proposed a model concerning possible roles of UVDDB and ubiquitylation in the damage recognition process in NER.
In vivo dynamics of XPC were examined by the FRAP (flumesmnce recovely after photobleaching) technique using a stably transformed cell line expressing GFP-tagged XPC. We observed immobilization of GFPXPC after UV-irradiation of cells, which showed a biphasic UV-dose dependency. SiRNA knockdown and overexpression of DDB2 indicated that the first step immobilization observed after relatively low doses (5-10 J/m^2) of UV depended on UV-DDB. Our results suggest that the UV-DDB-mediated damage recognition pathway may be predominant when only a small number of UV photolesions are induced.
To investigate biological meanings of the reversible ubiquitylation XPC induced by UV-irradiation, we screened siRNA libraries for enzymes involved in deubiquitylation of XPC. A candidate enzyme has been obtained, which is now under investigation. In addition, both thymine DNA glycosylase (TDG) and XPC have been known to be SUMOylated. Structural basis of the functional modulation of TDG by SUMOylation could be revealed by the X-ray crystal structures of SUMOylated TDG. We also determined SUMOylation sites in XPC and mutations at these sites caused partial NER defects in vivo.
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[Journal Article] Mutagenic radioadaptation in a human lymphoblastoid cell line2008
Author(s)
Yatagai, F., Umehayashi, Y., Homma, M., Sugasawa, K., Takayama, Y., Hanaoka, F
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Journal Title
Mutat. Res 638
Pages: 48-55
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] In vivo destabilization and functional defects of the xeroderma pigmentosum C protein caused by a pathogenic missense mutation2007
Author(s)
Yasuda, G., Nishi, R., Watanabe, E., Mori, T., lwai, S., Orioli, D., Stefanini, M., Hanaoka, F., Sugasawa, K
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Journal Title
Mal. CelL BioL 27
Pages: 6606-6614
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Crystal structure of SUMO-3-modified thymine-DNA glycosylase2006
Author(s)
Baba, D., Maita, N., Jce,J.-G., Uchimura, Y., Saitoh, H., Sugasawa, K., Hanaoka, F., Tochio, H., Hiroaki, H., and Shirakawa, M
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Journal Title
J. MoL BioL 359
Pages: 137-147
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The DNA repair-ubiquitin associated HR23 proteins are constituents of neuronal inclusions in specific neurodegenerative disorders without hampering DNA repair2006
Author(s)
Bergink, S., Severijnen, L.-A., Wijgers, N., Sugasawa, K., Yousaf, H., Kros, J. M., van Swieten, J., Oostra, B. A., Hoeijmakers, J. H. J., Vermeulen, W., Willernsen, R
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Journal Title
Neurobial Dis 23
Pages: 708-716
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex2005
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Chu, G., Mori, T., Iwai, S., Tanaka, K., Tanaka, K., Hanaoka, F
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Crystal structure of thymine DNA glycosylase conjugated to SUMO-12005
Author(s)
Baba, D., Maita, N., lee, J.-G., Uchimura, Y., Saitoh, H., Sugasawa, K., Hanaoka, Tochio, H., Hiroaki, H., Shirakawa, M
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Journal Title
Nature 435
Pages: 979-982
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 protein2005
Author(s)
Nishi, R., Okuda, Y., Watanabe, E., Mori, T., lwai, S., Masutani, C., Sugasawa, K., Hanaoka, F
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Journal Title
Mot Cell Biol 25
Pages: 5664-5674
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Signal transducer and activator of transcription 3 is a key regulator of keratinocyte survival and proliferation following UV irradiation2005
Author(s)
Sano, S., Chan, K. S., Kira, M., Kalaoka, K., Takagi, S., Tarutani, M., Itami, S., Kiguchi, K., Yokoi, M., Sugasawa, K., Mori, T., Hanaoka, F., Takeda, J., DiGiovanni, J
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Journal Title
Cancer Res 65
Pages: 5720-5729
Description
「研究成果報告書概要(欧文)」より
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[Presentation] In vivo dynamics and regulation of nucleotide excision repair factors2008
Author(s)
Nishi, R., Alekseev, S., Dinant, C., Hoogstraten, D., Hoeijmakers, J. H. J., Vermeulen, W., Hanaoka, F., Sugasawa, K
Organizer
Workshop on DNA Recombination and Chromosomal Rearrangement 2007, and The 19th Workshop on DNA Replication and Segregation
Place of Presentation
Izu, Japan
Year and Date
20080504-07
Description
「研究成果報告書概要(欧文)」より
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[Presentation] In vivo dynamics and regulation of nucleotide excision repair factors2007
Author(s)
Nishi, R., Alekseev, S., Dinant, C., Hoogstraten, D., Hoeijmakers, J. H. J., Vermeulen, W., Hanabka, F., Sugasawa, K
Organizer
BMB2007
Place of Presentation
Yokohama, Japan
Year and Date
20071211-15
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Structural Basis of functional modulation by SUMO of DNA repair and transcription factors2007
Author(s)
hirakawa, M., Baba, D., Arita, K., Tateishi, Y., Saitoh, H., Sugasawa, K., Nakai, A., Sekiyama, N., Tochio, T., Ariyoshi, M., Hiroaki, H
Organizer
BMB2007
Place of Presentation
Yokohama, Japan
Year and Date
20071211-15
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Damage recognition factors and their functions in nucleotide excision repair2006
Author(s)
Sugasawa, K., Yasuda, G., Watanabe, E., Nishi, R., Hanaoka, F
Organizer
The Molecular Biology Society of Japan Forum 2006
Place of Presentation
Nagoya, Japan
Year and Date
20061206-08
Description
「研究成果報告書概要(欧文)」より
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[Presentation] A molecular mechanism underlying recognition of UV-induced DNA damage2006
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 20th IUBMB International Congress of Biochemistry and Molecular Biology and the 11th FAOBMB Comgress
Place of Presentation
Kyoto, Japan
Year and Date
20060618-23
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Biochemical studies on UV-DDB using nucleosomal DNA substrates2006
Author(s)
Yasuda, T., Sugasawa, K., Yasuda, G., Nishi, R., lwai, S., Shiomi, T., Hanaoka, F
Organizer
The 20th IUBMB International Congress of Biochemistry and Molecular Biology and the 11th FAOBMB Comgress
Place of Presentation
Kyoto, Japan
Year and Date
20060618-23
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Roles of ubiquitylation in recognition of UV-induced DNA damage2006
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
Meeting on DNA Repair: from Molecular Mechanism to Human Disease
Place of Presentation
Noordwijkerhout, The Netherlands
Year and Date
20060402-07
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Damage recognition mechanism and roles of ubiquitylation in nucleotide excision repair2005
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 28th Annual Meeting of the Molecular Biology Society of Japan
Place of Presentation
Fukuoka, Japan
Year and Date
20051207-10
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Structural and functional modulation of DNA repair enzyme TDG by SUMO2005
Author(s)
Baba, D., Maita, N., lee, J.-G., Uchimura, Y., Saitoh, H., Sueasawa, K., Hanaoka, F., Tochio, T., Hiroaki, H., Shirakawa, M
Organizer
The 28th Annual Meeting of the Molecular Biology Society oflapan
Place of Presentation
Fukuoka, Japan
Year and Date
20051207-10
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Structure and molecular recognition of the UBL-UBA domains2005
Author(s)
Shirakawa, M., Fujiwara, K., Ohno, A., Kobayashi, H., Sugasawa, K., Hanaoka, F., Tenno, T., Tochio, T., Hiroaki, H
Organizer
The 28th Annual Meeting of the Molecular Biology Society oflapan
Place of Presentation
Fukuoka, Japan
Year and Date
20051207-10
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Functional defects of a mutant XPC containing a pathogenic single amino acid substitution2005
Author(s)
Yasuda, G., Nishi, R., Watanabe, F., Mori, T., lwai, S., Sugasawa, K., Hanaoka, F
Organizer
The 28th Annual Meeting of the Molecular Biology Society ofJapan
Place of Presentation
Fukuoka, Japan
Year and Date
20051207-10
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Roles of ubiquitylation in mammalian nucleotide excision repair2005
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 78th Annual Meeting of the Japanese Biochemical Society Kobe
Place of Presentation
Japan
Year and Date
20051019-22
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Functional defects of the mutant XPC protein harboring a single amino acid substitution2005
Author(s)
Yasuda, G., Nishi, R., Watanabe, E., Mori, T., lwai, S., Sugasawa, K., Hanaoka, F
Organizer
The 77th Annual Meeting of the Genetics Society oflapan Tokyo
Place of Presentation
Japan
Year and Date
20050927-29
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Damage recognition mechanism and roles of ubiquitylation in nucleotide excision repair2005
Author(s)
Sugasawa, K., Saijo, M., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 64th Annual Meeting of the Japanese Cancer Association
Place of Presentation
Sapporo
Year and Date
20050914-16
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Ubiquitylation of damage recognition factors involved in nucleotide excision repair2005
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 9th International Conference on Environmental Mutagens San Francisco
Place of Presentation
U.S.A
Year and Date
20050903-08
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Roles of ubiquitylation in mammalian nucleotide excision repair2005
Author(s)
Sugasawa, K., Okuda, Y., Saijo, M., Nishi, R., Matsuda, N., Tanaka, K., Tanaka, K., Hanaoka, F
Organizer
The 58th Annual Meeting of die Japan Society for Cell Biology
Place of Presentation
Saitama
Year and Date
20050615-17
Description
「研究成果報告書概要(欧文)」より
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