哺乳動物での活性酸素によるDNA損傷の修復とその
欠損の影響
著者
安井 明
晴乳動物での活性酸素によるDNA損傷の修復とその欠損の影響
課題番号 13480162
平成1 3年度∼平成1 5年度科学研究費補助金･基盤研究(B) (2)
研究成果報告書
平成17年2月
研究代表者
安井 明
(東北大学加齢医学研究所 教授)
はしがき
平成1 3年度から平成1 5年度にわキって交付された科学研究費補助金･基盤B
｢晴乳動物での活性酸素によるDNA損傷の修復とその欠損の影響｣の研究に依
って得られた成果を報告する｡活性酸素で生じる塩基損傷とりわけ致死的なチ
ミングリコール等を取り除き塩基除去修復を開始するマウスNTHlのノックア
ウトマウスの作成とその影響の解析､ NTHlをサポートする修復活性の同定｡
大腸菌のnthlをバックアップするendoVIIIグリコシラーゼのヒトホモローグ
NEILI, NEIL王Ⅰ及びNEILIIIの発見とその機能解析､さらにヒト細胞での単鎖
切断の修復を解析する実験系(ⅩPA-UVDEの系)の開発とそれを用いた修復プ
ロセスの発見､また､レーザーを用いた種々の活性酸素損傷をヒト細胞の核に
局所的に作成する方法の開発とそれを用いた修復プロセスのリアルタイム解析､
など､これまでに長く課題とされていた疑問に答え､将来の発展的研究を約束
する研究の展開があった｡
研究代表者及び分担者:
研究代表者:安井 明 (東北大学･加齢医学研究所･教授)
研究分担者:高尾 雅 (東北大学･加齢医学研究所･講師)
研究経費:
交付決定額
直接経費 間接経費 合計平成1 3年度
平成1 4年度
平成1 5年度
合計
5,100 0 4,700 0 4,700 0 14,500 0 5,100 4,700 4,700 14,500研究発表:
ア 学会誌等
(1) Okano S, Lan L, Tomkinson AE, and Yasui A. Translocation of XRCCl and DNA
日gase =a from centrosdomes to chromosomes in response to DNA damage､in
mitotic human ce"S. Nuc/e/'c Ac/'ds Res. In press.
(2) Wilson TM, Vaisman A, Martomo SA, Su"ivan P, Lan L Hanaoka F, Yasui A, Wbodgate R, and Gearhart PJ. MSH2-MSH6 stimu一ates DNA polymerase ll,
suggesting a roIe for A-T mutation in jmmunoglobulin genes. J. Exp. Med. In PreSS･
(3) Jans J, Schul W, Sert Y-G, Rijksen Y, Rebel H, EkerAPM, Nakajjma S, van Steeg
H, de GrLjijトFR, Yasui A, Hoeijmakers JHJ, van der Horst GTJ. PoweげuJ skin
cancer protection by a CPD-photolyase transgene. Cull. Bjo/ogy, 15, 1-20, 2005. (4) Zhang QM, Yonekura S, Takao M, Yasui A, Sugiyama H, and Yonei S. DNA
glycosylase activities for thymine residues oxidized in the methyl group are
functionsくJf the hNE暮Ll and hNTHl enzymes in human ceHs. DNA月epair, 4,
71-79, 2005.
(5) Nakajima S, Lan Li, Kanno S, Takao M, Yamamoto K, Eker APM, and Yasui A.
UV-induced DNA damage and to一erance for survival of nucleotide excision
repair-deficient human ce"S. J. BI'0/. Chem. 279, 46674-46677, 2004.
(6) Lan L, Nakajjma S, Oohata Y, Takao M, Okano S, Masutani M, Wi一son SH, and
Yasui A. /n situ analysis of repair processes for oxidative DNA damage in mammaJian ceHs. Proc. Nat/. Acad. Sci. USA. 101, 13738113743, 2004,
(7) Lan L. Hayashil T, Rokshana M. R, Nakajima S, Kanno S, Takao M, Matsunaga T, Yoshinq M, Ichikawa M, te Riete H, Tsuchiya S, Tanaka K, and Yasui A. Functional and physicaHnteractions between ERCCl and MSH2 complexes for resistance to c/'S-diammjnedichloroplatinum(lI) in mammalian ce"S. DNA Repa/'r3.
135-143, 2004.
(8) Filch ME, Nakajima S, Yasui A, and Ford JM. /n vJ'vo recruitment of XPC to UV-induced cyclobutane pyrimidine dimmers by the DDB2 gene product. J. BID/.
α7em 278, 46906-46910, 2003
(9) Okano S, Lan L, Caldecot K, Mori T, and Yasui A. Spatial and temporal assembly of the proteins involved in repalr Of single-strand breaks in human ce"S･ Mo/･ CeIJ･
Bio/. 23. 3974-3981. 2003.
(10) Takahashi H, Umeda N, Tsutumi Y, Fukumura R, Ohkaze H, Sujino M, van
der Horst, G, Yasui A, ]noue ST, Fujimori A, Ohhata T, Araki R, and Abe M. Mouse dexarmethasone･induced RAS protein 1 gene is expressed in a circadian rhythmic manner in the suprachiasmatjc nucleus. Mo/. BraI'n Resl 110, i-6, 2003･
(ll) Takao M, KannoS, Kobayashi K, Zhang QIM,, Yonej･S, vanderHorstG,and
Yasui A. A back-up glycosylase in Nthl-knockout mice is a functional Nei (endonuclease VHl) homologue. J. BID/. Chem･ 277, 42205-42213, 2002･
(12) Chigancas V, Batista LF, Brumattj G, Amarante-Mendes GP, YasuiA, Menck
CF. Photorepair of RNA polymerase arrest and apoptosis after ultraviolet
irradiation in normal and XPB deficient rodent ce"S. Gen Dea肋 Differ. 9,
109911107, 2002.
(13) Oster H, Yasui A, van der Hors,t GTJ, and Albrecht UI Disruption of mCrJQ
restores circadian rhythmicity in mPeI2 mutant mice. Genes Dev, 16, 2633-2638,
2002.
(14) AIbus H, Bonnefont X, Chaves I, YasuiA, DoczyJ, vander HorstGT, Meijer JH. Cryptochrome-deficient mice 一ack cjrcadjan electrical activity in the
suprachiasmatic nuclei. Curl BID/. 9, 1130-1133, 2002･
(15) Schul W, JansJ, Rijksen YMA, Klemann HM, EkerAPM, deWitJ, NikaidoO,
Nakajima S, Yasui A, Hoeijmakers JHJ, and van der Horst GTJ. Enhanced repair of cyclobutane pyrimidine djmers and improved UV resistance in photolyase
transgenic mice. EMBO J. 21 , 471 9-4729, 2002.
(16) Miiyabe I., Zhang QM., Kino K., Sugiyama H" Takao M･, YasujA" and Yonei
S. Identification of 5-formyluracil DNA g)ycosylase activity of human hNTHI
protein. Nuc/e/'c AcJ'ds月es. 30, 3454-3463, 2002･
(17) Takao M, Kanno S, Shiromoto T, Hasegawa R, lde H, ljkeda S, SarkerAH, Seki S, Xing JZ, Le XC, Weinfeld M Kobayashi K, Miyazaki J, Muijtjens M, Hoeijmakers JHJ, van der Horst G, and Yasui. A Novel nuclear and mitochondrial glycosylases revea)ed by disruption of the mouse Nthl gene encoding an endonuclease lII homologue for repalr Of thymine glycol. EMBO J. 21･ 3486-3493, 2002
(18) You YH, Lee DH, Yoon JH, Nakajima S, Yasui A, and Pfeifer GP･
Cyclobutane pyrimidine dimers are responsible for the vast majority of mutations induced by UVB irradiation in mammalian ce"S･ J Bjo/ Chem･ 276. 44688-44694,
2001.
(19) Nara, K., Nagashima, F., and Yasui, A. Highly elevated UV-induced mutation
frequency ln isolated Chinese hamster ceJJ fines defective in nucleotide excision
repalr and mismatch repalr PrOteins･ Cancer月es･ 61, 50-52, 2001 ･
(20) Matumoto, Y., Zhang, Q.M., Takao, M., Yasui, A., and Yonei, SI EscherJ'chia co//'Nth and human hNTHI DNA gIycosylases are involved in removal of 8-0Xoguanine from 810XOguanine/guanine mlSPair in DNA･ Nucleic Ac/'ds月es･ 29,
1975-1981, 2001.
(21) Okano, S., Kanno, S., Nakajima, S., and Yasui. A. Cellular responses and
repalr of sJngle-strand breaks introduced by UV damage endonucJease in mammalian ce"S. J. BJ'0/. Chem. 275, 32635-32641 , 2000.
(22) Chigansas, V., Miyaji, E.N., Muotri, A.R., Jacysyn, J.F., Amarante-Mendes,
G.P., Yasui, A., and Menck, C. F. M. Photorepalr Prevents UV-induced apoptosis in human cells expressJng the marsuplal photolyase gene. Cancer Res. 60,
2458-2463, 2000.
イ 口頭発表(国際会議での招待講演のみ)
1. Spatia暮and temporal assembly of the proteins involved in repair of sing一e-strand
breaks in human ce=S. Satoshi Okano, Li Lan, Toshio Mori, and Akira Yasui. First Japan-US DNA Repair Meeting, October 2002, Sendai.
2. Assembly of proteins and repaIr Process at OXidative DNA damage in mammalian celJs. Li Lan, Satoshi Nakajima, Yoshitugu Oohata, Toshio Mori, Satoshi Okano, Shin-ic-hiro Kanno, Masashi Takao and Akira Yasui. The 4th lnternationaJ 3R Symposium, November 9-13, 2003, Awajishjma Yumebutai.
3. Ce"ular responses to oxidative DNA damage in human ce"S. Li Lan, Satoshi Okano, Satoshi Nakajima and Akira Yasui. Second Japan-US DNA Repair Meeting, June 4-8, 2004, Honolulu. Hawaii.
4. /n s/'tu and real time analysIS Of repair pathways for base damage and single-strand
breaks in mammalian celts. Li Lan, Satoshi Nakajima, Satoshi Okano, S.H. Wilson
and Akira Yasui. ASM meeting: Conference on DNA repaFr and Mutagenesis,
November 14-20, 2004, Bermuda.
5. UV-induced DNA damage and toIerance for survival of nucleotide excision
repai卜deficient in human ceHs. Satoshi Nakajima, Li Lan, and Akira Yasui･ ASM
meeting: Conference on DNA repair and Mutagenesis. November 14120, 2004,
Bermuda.
6. /n s/'tu analysis of oxidative DNA damage repalr in human cells: Central roles of XRCCl in the responses of human ceHs to single-strand breaksI Li Lan, Satoshi Nakajima, Satoshi Okano and Akira Yasui. International Symposium on Genomic Maintenance and DNA Repair Network, January 26. 2005, Kyoto1
7. Dynamic Responses to Single-Strand<Breaks in Human CellsI Li Lan, Satoshi Okano,
Satoshi Nakajima and Akira Yasui. Keystone革ymposia; Genome lnstability and
Repair, March 15120, 2005, Taos, New Mexico.
