1999 年度実績報告書

染色体複製開始の制御機構

研究課題

研究課題/領域番号	08277101
研究機関	大阪大学
研究代表者	滝澤温彦大阪大学, 大学院・理学研究科, 助教授 (60154944)
研究分担者	升方久夫大阪大学, 大学院・理学研究科, 教授 (00199689) 四宮知行三菱化学生命化学研究所, 核機能プロジェクト, 主任研究員松影昭夫愛知県がんセンター研究所, 生物学部, 部長 (90019571) 片山勉九州大学, 薬学部, 助手 (70264059) 正井久雄東京大学, 医科学研究所・分子生物研究部, 助教授 (40229349)
キーワード	DNA複製 / MCM / ORC / CDC7 / CDC45 / 染色体 / DNAポリメラーゼ / 複製開始装置
研究概要	複製開始領域:四宮はショウジョウバエの複製開始点を1kb以下の解像度で調べ、開始点はAT含量の高い位置と一致し、核マトリックス結合部位によって挟まれていることを明らかにした。白髭は出芽酵母の第六染色体を用いて複製開始点の活性化制御機構について解析を行い、初期開始点と後期開始点ではヌクレオソームの配置が異なることを見いだした。複製開始タンパク質:升方は分裂酵母を用いてORCサブユニットOrp1が異なるC末領域を介して、Cdc18とMCMの複製開始点の結合に関わること、Cdc45の結合に新規因子が必要なことを明らかにした。滝澤はアフリカツメガエル卵無細胞複製系を用いてCdc45はDNA2本鎖を開きDNAポリメラーゼαを染色体上に結合させる過程に必須であり、DNAポリメラーゼεの染色体結合に必要であることを示した。正井は分裂酵母Cdc7キナーゼ複合体の遺伝学的な解析を行い、Cdc7キナーゼはMcm2,Rad3およびRad21と相互作用することを示した。またノックアウトマウスを用い、動物細胞のCdc7類似遺伝子がDNA複製と細胞増殖に必須であることを明らかにした。複製開始制御:田中は分裂酵母を用いて新たに同定したEso1がDNA複製複合体に局在し、新たに合成された姉妹染色体をその場で繋ぎとめると同時に、複製フォークの進行が阻害された時には障害をバイパスさせる因子であることを示した。片山はDNAポリメラーゼIIIによるDnaA不活性化の分子機構を解析し、同調培養細胞を用いて、複製周期と共役したDnaA蛋白質の活性フォームの変換が起こり複製開始後に不活性型に移行することを明らかした。複製関連遺伝子群発現制御:松影はショウジョウバエDREFのヒトホモログ(hDREF)のcDNAを単離し、hDREFタンパク質が特的なDNA配列に結合し、ヒトサイトメガロウイルスの極初期遺伝子の発現調節領域に存在するサイレンサーエレメントに結合する細胞性因子であることを証明した。

研究成果

(22件)

すべてその他

すべて文献書誌 (22件)

[文献書誌] Mimura, S.: "Central role for Cdc45 in establishing an initiation compex of DNA replication in Xenopus egg extracts"Genes Cells in press.
[文献書誌] Ohta, K.: "Action of a new mammalian DNA polymer ase inhibitor, sulfoquinovosl diacylglycerol"Biol. Pharm. Bull.. 22. 111-116 (1999)
[文献書誌] Takahashi, Y.: "Identification of three conserved regions in the DREF transcription factors from Drosophila melanogaster and Drosophila virilis"Nucleic Acids Res.. 27. 510-516 (1999)
[文献書誌] Hirose, F.: "Targeted expression of the DNA binding domain of Drosophila transcription factor DREF attenuates DNA replication"Mol. Cell. Biol.. 18. 6020-6028 (1999)
[文献書誌] Mizushima, Y.: "The cyanogenic glucoside, prenasin (D-mandelorritril-D-glucoside), is a novel inhibitor of DNA polymerase b"J. Biochem.. 126. 430-436 (1999)
[文献書誌] Yamaguchi, M.: "Ectopic expression of human p23 inhibits entry into S-phase and induces apoptosis in the Drosophila eye imaginal disc"Oncogene. 18. 6767-6775 (1999)
[文献書誌] Hayashi, Y.: "A binding site for transcription factor Granyhead/nuclear transcription factor-1 contributes to regulation"J. Biol. Chem.. 274. 35080-35088 (1999)
[文献書誌] Masai, H.: "Regulation of DNA replication duringsell sysle and environmental stresses"IUBMB Life, in press.
[文献書誌] Masai, H.: "Cdc7/Dbf4-related kinase complex as a molecular switch for initiation of DNA replication"Frontiers in Bioscience. 4. 834-840 (1999)
[文献書誌] Takada, T.: "A Fission yeast gane, him1+/dfp1+, encoding a regulatory subunit for Hsk1 kinase, play essential roles in S phase initiation"Mol. Cell. Biol.. 19. 5535-5547 (1999)
[文献書誌] Masai, H.: "Escherichia coil and Bacillus subtilis PriA proteins essential for recombination-dependent DNA replication : involvement of ATPase"Biochimie. 81. 847-857 (1999)
[文献書誌] Kumagai, H.: "A novel growth-and cell cycle-regulated protein, ASK, activates human Cdc7-related kinase and is essential for G1/S transition"Mol. Cell. Biol.. 19. 5083-5095 (1999)
[文献書誌] Johnstone, L.: "First the Cdk's, now the Ddk's"Trends in Cell Biology. 9. 249-252 (1999)
[文献書誌] Y. Okuno: "Clustered adenine/thymine stretches are essential for the function of a fission yeast replication orgin"Mol. Cell. Biol.. 19・10. 6699-6709 (1999)
[文献書誌] Y. Ogawa: "Association of fission yeast Orp1 and Mcm6 proteins with chromosomal replication origins"Mol. Cell. Biol.. 19・10. 7228-7236 (1999)
[文献書誌] Masuda, M.: "Genetic studies with the fission yeastSchizosaccharomyces pombe suggest involvement of Wee1, Ppa2, and Rad24 in induction of cell"J. Virol.. 74 (in press).
[文献書誌] Koichi Tanaka: "Fission Yeast Eso1p is required for establishing sister chromatid cohesion during S phase"Mol. Cell. Biol.. 20 (in press).
[文献書誌] Shirahige, K.: "The Cloning and characterization of CDC50 gane family in S. cerevisiae"Yeast. (in press).
[文献書誌] Takata, M.: "MutantDnaA prteins defective in opening of oriC, the origin of chromosomal DNA replication in Escherichia coil"Mol. Microbiol.. 35・2. 454-462 (2000)
[文献書誌] Kurokawa, K.: "Replication cycle-coordinated change of the adenine nusleotide-bound forms of DnaA Protein in Escherichia coli"ENBO J.. 18・23. 6642-6652 (1999)
[文献書誌] Guo, L.: "Isolation and characterization of novel cold-sensitive dnaA mutants of Escherichia coli"FEMS Microbiol. lett.. 176. 357-366 (1999)
[文献書誌] Katayama, T.: "Inactivation of Escherichia coli DnaA protein by DNA polymerase III, and negative regulations for initiation of chromosomal replication"Biochimie. 81・8. 835-840 (1999)

1999 年度 実績報告書

染色体複製開始の制御機構

研究代表者

滝澤 温彦 大阪大学, 大学院・理学研究科, 助教授 (60154944)

研究成果

[文献書誌] Mimura, S.: "Central role for Cdc45 in establishing an initiation compex of DNA replication in Xenopus egg extracts"Genes Cells in press.

[文献書誌] Ohta, K.: "Action of a new mammalian DNA polymer ase inhibitor, sulfoquinovosl diacylglycerol"Biol. Pharm. Bull.. 22. 111-116 (1999)

[文献書誌] Takahashi, Y.: "Identification of three conserved regions in the DREF transcription factors from Drosophila melanogaster and Drosophila virilis"Nucleic Acids Res.. 27. 510-516 (1999)

[文献書誌] Hirose, F.: "Targeted expression of the DNA binding domain of Drosophila transcription factor DREF attenuates DNA replication"Mol. Cell. Biol.. 18. 6020-6028 (1999)

[文献書誌] Mizushima, Y.: "The cyanogenic glucoside, prenasin (D-mandelorritril-D-glucoside), is a novel inhibitor of DNA polymerase b"J. Biochem.. 126. 430-436 (1999)

[文献書誌] Yamaguchi, M.: "Ectopic expression of human p23 inhibits entry into S-phase and induces apoptosis in the Drosophila eye imaginal disc"Oncogene. 18. 6767-6775 (1999)

[文献書誌] Hayashi, Y.: "A binding site for transcription factor Granyhead/nuclear transcription factor-1 contributes to regulation"J. Biol. Chem.. 274. 35080-35088 (1999)

[文献書誌] Masai, H.: "Regulation of DNA replication duringsell sysle and environmental stresses"IUBMB Life, in press.

[文献書誌] Masai, H.: "Cdc7/Dbf4-related kinase complex as a molecular switch for initiation of DNA replication"Frontiers in Bioscience. 4. 834-840 (1999)

[文献書誌] Takada, T.: "A Fission yeast gane, him1+/dfp1+, encoding a regulatory subunit for Hsk1 kinase, play essential roles in S phase initiation"Mol. Cell. Biol.. 19. 5535-5547 (1999)

[文献書誌] Masai, H.: "Escherichia coil and Bacillus subtilis PriA proteins essential for recombination-dependent DNA replication : involvement of ATPase"Biochimie. 81. 847-857 (1999)

[文献書誌] Kumagai, H.: "A novel growth-and cell cycle-regulated protein, ASK, activates human Cdc7-related kinase and is essential for G1/S transition"Mol. Cell. Biol.. 19. 5083-5095 (1999)

[文献書誌] Johnstone, L.: "First the Cdk's, now the Ddk's"Trends in Cell Biology. 9. 249-252 (1999)

[文献書誌] Y. Okuno: "Clustered adenine/thymine stretches are essential for the function of a fission yeast replication orgin"Mol. Cell. Biol.. 19・10. 6699-6709 (1999)

[文献書誌] Y. Ogawa: "Association of fission yeast Orp1 and Mcm6 proteins with chromosomal replication origins"Mol. Cell. Biol.. 19・10. 7228-7236 (1999)

[文献書誌] Masuda, M.: "Genetic studies with the fission yeastSchizosaccharomyces pombe suggest involvement of Wee1, Ppa2, and Rad24 in induction of cell"J. Virol.. 74 (in press).

[文献書誌] Koichi Tanaka: "Fission Yeast Eso1p is required for establishing sister chromatid cohesion during S phase"Mol. Cell. Biol.. 20 (in press).

[文献書誌] Shirahige, K.: "The Cloning and characterization of CDC50 gane family in S. cerevisiae"Yeast. (in press).

[文献書誌] Takata, M.: "MutantDnaA prteins defective in opening of oriC, the origin of chromosomal DNA replication in Escherichia coil"Mol. Microbiol.. 35・2. 454-462 (2000)

[文献書誌] Kurokawa, K.: "Replication cycle-coordinated change of the adenine nusleotide-bound forms of DnaA Protein in Escherichia coli"ENBO J.. 18・23. 6642-6652 (1999)

[文献書誌] Guo, L.: "Isolation and characterization of novel cold-sensitive dnaA mutants of Escherichia coli"FEMS Microbiol. lett.. 176. 357-366 (1999)

[文献書誌] Katayama, T.: "Inactivation of Escherichia coli DnaA protein by DNA polymerase III, and negative regulations for initiation of chromosomal replication"Biochimie. 81・8. 835-840 (1999)

1999 年度実績報告書

滝澤温彦大阪大学, 大学院・理学研究科, 助教授 (60154944)