DNA damage-induced cell death and DNA repair network

• Project/Area Number: 12143101
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: TANAKA Kiyoji 大阪大学, 大学院・生命機能研究科, 教授 (80144450)
• Co-Investigator(Kenkyū-buntansha): YASUI Akira 東北大学, 加齢医学研究所, 教授 (60191110) ; YAMAIZUMI Masaru 熊本大学, 発生医学研究センター, 教授 (70107093) ; SEKIGUCHI Mutsuo 福岡歯科大学, 名誉教授 (00037342) ; HANAOKA Fumio 大阪大学, 大学院・生命機能研究科, 教授 (50012670) ; OGAWA Hideyuki 岩手看護短期大学, 教授 (70028207)
• Project Period (FY): 2000 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥76,500,000 (Direct Cost: ¥76,500,000); Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000); Fiscal Year 2004: ¥14,500,000 (Direct Cost: ¥14,500,000); Fiscal Year 2003: ¥14,500,000 (Direct Cost: ¥14,500,000); Fiscal Year 2002: ¥23,000,000 (Direct Cost: ¥23,000,000); Fiscal Year 2001: ¥23,000,000 (Direct Cost: ¥23,000,000)
• Keywords: DNA damage / DNA repair / xeroderma pigmentosum / Cockayne syndrome / postreplication repair / knockout mice / nuclear matrix / transcription / DNA修復 / 損傷トレランス / 転写 / DNA複製 / ユビキチンリガーゼ / ノックアウトマウス / 機能的プロテオーム / 損傷乗り越え修復 / DNA損傷トレランス / 色素性乾皮症 / 紫外線 / DNA損傷 / 複製後修復 / 核マトリクス / DNA修復ネットワーク / 細胞死 / チミングリコール / 転写伸長 / XPA / NTH1 / リン酸化 / 突然変異

Research Abstract

2000〜2005年の研究成果に基づき研究成果報告書冊子を作成した。成果概要は以下のとおりである。RNAポリメラーゼIIによる転写をブロックし細胞死を誘発する転写鎖上のDNA損傷は、「転写と共役した修復」(TCR)により修復される。コケイン症候群(CS)はTCRを選択的に欠損し、その原因遺伝子産物であるCSA、CSBはTCRに必須であることが示唆される。CSAはDDB1、Roc1、Cullin4A、CSNと複合体を形成し、RNAポリメラーゼIIをユビキチン化するE3活性をもつことを見つけた。色素性乾皮症G群(XPG)患者にはCSを併発する場合がある。4種類のXPG変異マウスを作製し、そのC末端領域の欠損がCSの発症に関与することを明らかにした。「ゲノム全体の修復」(GGR)に関与するDDB2は、CSA同様に、DDB1、Roc1、Cullin4A、CSNと複合体を形成しユビキチンリガーゼ活性をもつこと、そのターゲットがXPCやDDB2自身であること、これらのユビキチン化がNERの促進に働くことを明らかにした。ヒト細胞核にレンズを通してレーザーを照射し、局所的に種々の活性酸素による損傷を作る技術を開発し、塩基損傷や単鎖切断の修復蛋白がリアルタイムで集積、解離するin situ修復プロセスを明らかにした。TLSに関与するヒトRAD18ユビキチンリガーゼや色素性乾皮症バリアント群の原因遺伝子産物であるDNAポリメラーゼエータ(Pol eta)を発見し、それらのTLS機能を明らかにした。マウスRAD18やPol eta欠損マウスを作成し、TLS機能の個体レベルでの役割の解析に重要な手段を提供できるようにした。また、ワークショップを主催し、計画研究班員は成果を発表した。研究総括のため、最後の班会議を開催した。2006年10月には、文部科学省で研究終了ヒアリングを受け、Aの評価を得た。

Research Products

• [Journal Article] Disruption of mouse XAB2 gene involved in pre-mRNA splicing, transcription and transcription-coupled DNA repair results in preimplantation lethality (2005)
  • Author(s): Rie Yonemasu
  • Journal Title: DNA Repair 4 Pages: 479-491
• [Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin lipase complex (2005)
  • Author(s): Kaoru Sugasawa
  • Journal Title: Cell 121 Pages: 387-400
• [Journal Article] Translocation of XRCC1 and DNA ligase IIIa from centrosomes to chromosomes in response to DNA damage (2005)
  • Author(s): S.Okano
  • Journal Title: Nucl.Acid Res. 33 Pages: 422-429
• [Journal Article] Differential regulation of Rad18 through Rad6-dependent mono- and polyubiquitination (2005)
  • Author(s): K.Watanabe
  • Journal Title: J.Biol.Chem. 280 Pages: 515-524
• [Journal Article] Centrin 2 stimulates nucleotide excision repair by interacting with xeroderma pigmentosum group C protein. (2005)
  • Author(s): R.Nishi
  • Journal Title: Mol.Cell.Biol. 13 Pages: 5664-5674
• [Journal Article] Severe growth retardation and short life span of double mutant mice lacking Xpa and Exon 15 of XPG (2005)
  • Author(s): Naoko Shiomi
  • Journal Title: DNA Repair 4 Pages: 351-357
• [Journal Article] Disruption of mouse XAB2 gene involved in pre-mRNA splicing, transcription and transcription-coupled DNA repair results in preimplantation lethality. (2005)
  • Author(s): Rie Yonemasu., Mitsuyoshi Minami., Yoshimichi Nakatsu., Masayo Takeuchi., Isao Kuraoka, Yoichi Matsuda., Yujiro Higashi., Hisato Kondoh, Kiyoji Tanaka
  • Journal Title: DNA Repair 4 Pages: 479-491
• [Journal Article] Complete absence of Cockayne syndrome group B gene product gives rise to UV-sensitive syndrome but not Cockayne syndrome. (2004)
  • Author(s): Katsuyoshi Horibata., Yuka Iwamoto, Isao Kuraoka., Nicolaas G.J., Jaspers., Masahiro Kurimasa., Mitsuo Oshimura., Masamitsu Ichihashi, Kiyoji Tanaka.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 15410-15415
• [Journal Article] In situ analysis of repair processes for oxidative DNA damage in mammalian cells. (2004)
  • Author(s): Lan., L., Nakajima., S., Oohata, Y., Takao., M., Okano, S., Masutani., M., Wilson., S.H., Yasui, A.
  • Journal Title: Proc.Natl.Acad.Sci.USA 101 Pages: 13738-13743
• [Journal Article] UV Light-induced DNA Damage and Tolerance for the Survival of Nucleotide Excision Repair-deficient Human Cells (2004)
  • Author(s): Satoshi Nakajima., Li Lan, Shin-ichiro Kannno, Masashi Takano, Kazuo Yamamoto, Andre P.M.Eker., Akira Yasui
  • Journal Title: The Journal of Biological Chemistry 279 Pages: 46674-46677
• [Journal Article] Rad18 guides pol eta to replication stalling sites through physical interaction and PCNA monoubiquitination. (2004)
  • Author(s): Kenji Watanabe, Satoshi Tateishi., Michiko Kawasuji, Toshiki Tsurimoto., Hirokazu Inoue, Masaru Yamaizumi.
  • Journal Title: The EMBO Journal 23 Pages: 3886-3896
• [Journal Article] UV-induced ubiquitylation of XPC protein mediated by UV-DDB-ubiquitin ligase complex.
  • Author(s): Kaoru Sugasawa., Yuki Okuda., Masafumi Saijo., Ryotaro Nishi., Noriyuki Matsuda., Gilbert Chu., Toshio Mori., Shigenori Iwai., Keiji Tanaka., Kiyoji Tanaka, Fumio Hanaoka.
  • Journal Title: Cell (in press.)
• [Publications] Isao Kuraoka: "Effects of endogeneous DNA base lesions on transcription elongation by mammalian RNA polymerase II : Implications for transcription-coupled DNA repair and transcriptional mutagenesis."J.Biological Chemistry. 278. 7294-7299 (2003)
• [Publications] Regina Groisman: "The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage."Cell. 113. 357-367 (2003)
• [Publications] Satoshi Okano: "Spatial and temporal assembly of the proteins involved in repair of single-strand breaks in human cells."Molecular and Cellular Biology. 23. 3974-3981 (2003)
• [Publications] Li Lan: "Functional and physical Interactions between ERCC1 and MSH2 complexes for resistance to cis-diamminedichloroplatinum(II) in mammalian cells."DNA Repair. 3. 135-143 (2004)
• [Publications] Satoshi Tateishi: "Enhanced genomic instability and defective postreplication repair in RAD18 knockout mouse embryonic stem cells."Molecular and Cellular Biology. 23. 474-487 (2003)
• [Publications] Yuichiro Shimizu: "Xeroderma pigmentosum group C protein interacts physically and functionally with thymine DNA glycosylase."EMBO J.. 22. 164-173 (2003)
• [Publications] Regina Groisman: "The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage"Cell. 113. 357-367 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Shinya Kamiuchi: "Translocation of Cockayne syndrome group A protein to the nuclear matrix : possible relevance to transcription-coupled DNA repair"Proc. Natl. Acad. Sci., USA. 99. 201-206 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Machiko Murai: "Early postnatal ataxia and abnormal cerebellar development in mice lacking xeroderma pigmentosum group A and the Cockayne syndrome group B DNA repair genes"Proc. Natl. Acad. Sci., USA. 98. 13379-13384 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Masashi Takao: "Novel nuclear and mitochondrial glycosylases revealed by disruption of the mouse Nth1 gene encoding an endonuclease III homologue for repair of thymine glycol"EMBO J.. 21. 3486-3493 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Satoshi Tateishi: "Dysfunction of human Radl8 results in defective postreplication repair and hypersensitivity to multiple mutagens"Proc. Natl. Acad. Sci. USA. 97. 7927-7932 (2000)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Satoshi Tateishi: "Enhanced genomic instability and defective postreplication repair in RAD 18-knockout mouse-embryonic stem cells"Mol. Cell. Biol.. 23. 474-481 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Regina Groisman: "The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage"Cell. 113. 357-367 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Shinya Kamiuchi: "Translocation of Cockayne syndrome group A protein to the nuclear matrix : possible relevance to transcription-coupled DNA repair"Proc. Natl. Acad. Sci., USA. 99. 201-206 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Machiko Murai: "Early postnatal ataxia and abnormal cerebellar development in mice lacking xeroderma pigmentosum group A and the Cockayne syndrome group B DNA repair genes"Proc. Natl. Acad.Sci., USA. 98. 13379-13384 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Masashi Takao: "Novel nuclear and mitochondrial glycosylases revealed by disruption of the mouse Nth1 gene encoding an endonuclease III homologue for repair of thymine glycol"EMBO J.. 21. 3486-3493 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Satoshi Tateishi: "Dysfunction of human Rad18 results in defective postreplication repair and hypersensitivity to multiple mutagens"Proc. Natl. Acad. Sci. USA. 97. 7927-7932 (2000)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Satoshi Tateishi: "Enhanced genomic instability and defectivepostreplication repair in RAD18・knockout mouse-embryonic stem cells"Mol. Cell. Biol.. 23. 474-481 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Yoshimichi Nakatsu: "XAB2, a novel tetratricopeptide repeat protein, involved in transcription-coupled DNA repair and transcription"J. Biol. Chem.. 275. 34931-34937 (2000)
• [Publications] Shinya Kamiuchi: "Translocation of Cockayne syndrome group A protein to the nuclear matrix : possible relevance to transcription-coupled DNA repair"Proc. Natl. Acad. Sci., USA. 99. 201-206 (2002)
• [Publications] Takao M: "A back-up glycosylase in Nth1-knockout mice is a functional Nei (endonuclease VIII) homologue"J. Biol. Chem.. 277. 42205-42213 (2002)
• [Publications] Takao M: "Novel nuclear and mitochondrial glycosylases revealed by disruption of the mouse Nth1 gene encoding an endonuclease III homologue for repair of thymine glycol"EMBO J.. 21. 3486-3493 (2002)
• [Publications] Tateishi S: "Dysfunction of human Rad18 results in defective postreplication repair and hypersensitivity to multiple mutagens"Proc. Natl. Acad. Sci. USA. 97. 7927-7932 (2000)
• [Publications] Tateishi S: "Enhanced genomic instability and defectivepostreplication repair in RAD 18-knockout mouse-embryonic stem cells"Mol. Cell. Biol.. 23. 474-481 (2003)
• [Publications] Shinya Kamiuchi: "Translocation of Cockayne syndrome group A protein to the nuclear matrix : possible relevance to transcription-coupled DNA repair"Proc. Natl. Acad. Sci., USA. 99. 201-206 (2002)
• [Publications] Nara, K.: "Highly elevated UV-induced mutation frequency in isolated Chinese hamster cell lines defective in nucleotide excision repair and mismatch repair proteins"Cancer Research. 61. 50-52 (2001)
• [Publications] Kishi, H.: "Osmotic Shock Induces G1 Arrest through p53 Phosphorylation at Ser33 by Activated p38MAPK without Phosphorylation at Ser15 and Ser20"J. Biol. Chem.. 276. 39115-39122 (2001)
• [Publications] Sugasawa, K.: "A molecular mechanism of DNA damage recognition by the xeroderma pigmentosum group C protein complex"DNA Repair. 1. 95-107 (2002)
• [Publications] Wakasugi, M.: "DDB accumulates at DNA damage sites immediately after UV irradiation and directly stimulates nucleotide excision repair"J. Biol. Chem.. 277. 1637-1640 (2001)
• [Publications] Shiomi, N.: "Disruption of Xpg increases spontaneous mutation frequency, particularly A : T-to-C : G transversion"Mutat. Res.. 487. 127-135 (2001)
• [Publications] Nakatsu,Y.: "XAB2, a novel tetratricopeptide repeat protein, involved in transcription-coupled DNA repair and transcription"J.Biol.Chem.. 275. 34931-34937 (2000)
• [Publications] Okano,S.: "Cellular responses and repair of single-strand breaks introduced by UV damage endonuclease in mammalian cells"J.Biol.Chem.. 275. 32635-32641 (2000)
• [Publications] Tateishi,S.: "Dysfunction of human Rad18 results in defective postreplication repair and hypersensitivity to multiple mutagens"Proc.Natl.Acad.Sci.USA. 97. 7927-7932 (2000)
• [Publications] Sugasawa,K.: "A multi-step damage recognition mechanism for global genomic nucleotide excision repair"Genes Dev.,. (in press).
• [Publications] Masutani,C.: "Mechanisms of accurate translesion synthesis by human DNA polymerase n"EMBO J.. 19. 3100-3109 (2000)
• [Publications] Wakasugi,M.: "DDB stimulates the excision of cyclobutane pyrintidine dimers in vitro in concert with XPA and RPA"J.Biol.Chem.. (in press).