1998 Fiscal Year Annual Research Report

染色体複製開始の制御機構

Research Project

Project/Area Number	08277101
Research Category	Grant-in-Aid for Scientific Research on Priority Areas (A)
Research Institution	Osaka University
Principal Investigator	滝澤温彦大阪大学, 大学院・理学研究科, 助教授 (60154944)
Co-Investigator(Kenkyū-buntansha)	白髭克彦奈良先端科学技術大学院大学, バイオサイエンス研究科, 助手 (90273854) 正井久雄東京大学, 医科学研究所分子生物学研究部, 助教授 (40229349) 升方久夫大阪大学, 大学院・理学研究科, 助教授 (00199689) 四宮知行三菱化学生命科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)
Keywords	染色体複製 / 複製開始領域 / 複製開始装置 / 複製開始制御 / DNA複製関連遺伝子
Research Abstract	複製開始領域の同定・単離とその特性の解析:升方は分裂酵母を用いてORCが複製開始点に結合していること、MCMが細胞周期特異的に複製開始点に結合することを示した。四宮はショウジョウバエ染色体上の複製開始領域の詳細な構造を調べ、多数の開始点が不均一に分布し、いずれの開始点もAT含量の高い配列の近傍に存在することを明らかにした。白髭は出芽酵母第6染色体上にあるマルチレプリコンの複製開始の制御機構がDNA傷害や複製チェックポイント機構と密接な連関があることを明らかにした。複製開始領域と複製開始複合体タンパク質の相互作用:升方はORCを含む免疫沈降画分が分裂酵母ARS断片に特異的に結合することを発見した。複製開始に関わる因子の同定とその機能の解析:滝澤はアフリカツメガエル卵無細胞複製系を用いてXCdc45がDNAポリメラーゼαの染色体上への結合に必須な因子であり、XCdc45の染色体への結合はS期CDKの制御下にあることを明らかにした。正井は分裂酵苺とヒトのCdc7キナーゼ活性制御サブユニットの機能について検討し、これらサブユニットがキナーゼの活性に必須であり、DNA複製阻害時のチェックポイント制御などにも関与することを明かにした。複製開始の制御機構:田中は分裂酵母を用いて複製開始善複合体と相互作用する因子Spt1の遺伝子と強い遺伝学的関係を示すcdc24+遺伝子産物はPCNAやRFCと複合体を形成しDNA複製の進行と完了に必須の役割を果たす因子であることを明らかにした。片山はDNAポリメラーゼIIIによるDnaA不活性化(RIDA)の分子機構を解析し、複製中間体上で両者の蛋白質が相互作用することを示唆する結果を得た。また複製開始を促進する新規因子ActBはDnaAと直接相互作用することを明らかにした。複製関連遺伝子群の発現制御機構:松影はDREFを複眼原基で過剰発現するハエを用いてDREF遺伝子はE2F遺伝子の遺伝学的に上位で機能していることを明らかにした。またDREFの発現制御下にあるショウジョウバエのMCM3、6、7のcDNAを新たにクローン化した。

Research Products
(29 results)

All Other

All Publications (29 results)

[Publications] Mimura,S.: "Xenopus Cdc45-dependent loading of DNA polymerase α onto chromatin under the control of S-phase cdk." EMBO,J.17. 5699-5707 (1998)
[Publications] Katayama,T.: "The initiator function of DnaA protein is negatively regulated by the sliding clamp of the E.coli chromosomal replicase." Cell. 94(1). 61-71 (1998)
[Publications] Kurokawa,K.: "A stimulation factor for hydrolysis of ATP bound to DnaA protein,the initiator of chromosomal DNA replication in Escherichia coli." Biochem.Biophys.Res.Commun.243(1). 90-95 (1998)
[Publications] Mizushima,T.: "Site-directed mutational analysis for the ATP binding of DnaA protein: functions of two conserved amino acids (Lys-178 and Asp-235)located" J.Biol.Chem.273(33). 20847-20851 (1998)
[Publications] Sawado,T.: "The DNA replication-related element (DRE)/ DRE-binding factor system is a transcriptiona regulator ofthe Drosophila E2F gene." J.Biol.Chem.273. 26042-26051 (1998)
[Publications] Sawado,T.: "dE2F, a novel E2F-family transcription factor in Drosophila melanogaster." Biochem.Biophys.Res.Commun.251. 409-415 (1998)
[Publications] C.Obuse: "Interaction of transcription factor YY1 with a replication-enhancing element,REE1,in an autonomously replicating human chromosome" Nucle.Acids Res.1. 253-253 (1998)
[Publications] Sasaki,T.: "Specification of Initation Regions during Embryogenesis in the 65 kilobase DNA pol α-dE2F Locus of Drosophila melanogaster." Mol.Cel.Biol.,. 19,1. 547-555 (1999)
[Publications] Y.Ogawa,T: "Association of autonomous replication activity with replication origins in a human chromosome." Exp.Cell Res.243. 50-58 (1998)
[Publications] Horie,S.: "The Schizosaccharomyces pombe mei4+ gene encodes a meiosis-specific transcription factor containing a forkhead" Mol.Cell.Biol.18. 2118-2129 (1998)
[Publications] Tahara,S.: "Functional domains of Rep2,a transcriptional activator subunit for Res2-Cdc10,controlling the cell cycle“start"." Mol.Biol.Cell.9. 1577-1588 (1988)
[Publications] Shirahige,K: "Regulation of DNA replication origins during cell-cycle progression." Nature. 395. 618-621 (1998)
[Publications] Horie,T.: "A 5'-monophosphate form of bredinin selectively inhibits the activities of mammalian DNA polymerases in vitro." Intn.J.Mol.Med.1. 83-90 (1998)
[Publications] Ohta,K.: "Sulfoquinovosyldiacylglycerol,KM043,a new potent inhibitor of eukaryotic DNA polymerases and HIV-reverse transcriptase type I from" Gigartina tenella.Chem.Pharm.Bull.46. 684-686 (1998)
[Publications] Mizushina,Y.: "The inhibitory effect of novel triterpenoid compounds,fomitellic acids, on DNA polymerase b." Biochem.J.330. 1325-1332 (1998)
[Publications] Mizushina,Y.: "The biochemical inhibition mode of bredinin-5'-monophosphate on DNA polymerase b.Biochim." Biophys.Acta.1403. 5-11 (1998)
[Publications] Ohno,K.: "cDNA cloning and expression during development of Drosophila melanogaster MCM3,MCM6 and MCM7." Gene. 217. 177-186 (1998)
[Publications] Mizushina,Y.: "A mushroom fruiting body-inducing substance inhibits activities of replicative DNA polymerases." Biochem.Biophys.Res.Commun.249. 17-22 (1998)
[Publications] Ohtoshi,A: "Two recessive modes of growth inhibitionby exogenously introduced mutant genes: Analysis of mutant CDC28 and CDC7 genes" J.Biochem.Mol.Biol.Biophys.1. 253-263 (1998)
[Publications] Kim,J.M.: "Growth regulation ofthe expression of mouse cDNA and gene encoding a serine/threonine kinase related toS.cerevisiae CDC7 essential for" J.Biol.Chem.273. 23248-23257 (1998)
[Publications] Jonstone,L.: "A Cdc7-Dbf4 kinase activity is conserved from yeast to human" Progress in Cell Cycle Research. Vol.4(in press). (1999)
[Publications] 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)
[Publications] Mizushina,Y.: "Studies on inhibotors of mammalian DNA polymerase a and b. Sulfolipids from a pteridophyte,Athyrium niponicum." Biochem.Pharmacol.55. 537-541 (1998)
[Publications] Togashi,H.,: "4-Hydroxy-17-methylincisterol,and inhibitor of DNA polymerase-a activity and the growth of human cancer cell in vitro." Biochem.Pharmacol.56. 583-590 (1998)
[Publications] Tanaka,H.,: "Fission yeast Cdc24 is a replication factor C- and proliferating cell nuclear antigen-interacting factor essential for S phase completion." Mol.Cell.Biol.,. 19. 1038-1048 (1999)
[Publications] 片山勉: "DNAポリメラーゼの新機能:染色体複製開始の負の制御" 蛋白質核酸酵素. (印刷中). (1999)
[Publications] 滝澤温彦: "MCM/P1ファミリー蛋白質とDNA複製のライセンス化" 実験医学. 16巻8号. 61-66 (1998)
[Publications] 白髭克彦: "複製開始領域の構成と染色体構築-出芽酵母の場合" 実験医学. 16. 31-40 (1998)
[Publications] 白髭克彦: "酵母染色体のマルチレプリコン構造とチェックポイント制御細胞周期研究の新しい展開(田矢,野島編) (印刷中)" 羊土社,

1998 Fiscal Year Annual Research Report

染色体複製開始の制御機構

Principal Investigator

滝澤 温彦 大阪大学, 大学院・理学研究科, 助教授 (60154944)

Research Products

[Publications] Mimura,S.: "Xenopus Cdc45-dependent loading of DNA polymerase α onto chromatin under the control of S-phase cdk." EMBO,J.17. 5699-5707 (1998)

[Publications] Katayama,T.: "The initiator function of DnaA protein is negatively regulated by the sliding clamp of the E.coli chromosomal replicase." Cell. 94(1). 61-71 (1998)

[Publications] Kurokawa,K.: "A stimulation factor for hydrolysis of ATP bound to DnaA protein,the initiator of chromosomal DNA replication in Escherichia coli." Biochem.Biophys.Res.Commun.243(1). 90-95 (1998)

[Publications] Mizushima,T.: "Site-directed mutational analysis for the ATP binding of DnaA protein: functions of two conserved amino acids (Lys-178 and Asp-235)located" J.Biol.Chem.273(33). 20847-20851 (1998)

[Publications] Sawado,T.: "The DNA replication-related element (DRE)/ DRE-binding factor system is a transcriptiona regulator ofthe Drosophila E2F gene." J.Biol.Chem.273. 26042-26051 (1998)

[Publications] Sawado,T.: "dE2F, a novel E2F-family transcription factor in Drosophila melanogaster." Biochem.Biophys.Res.Commun.251. 409-415 (1998)

[Publications] C.Obuse: "Interaction of transcription factor YY1 with a replication-enhancing element,REE1,in an autonomously replicating human chromosome" Nucle.Acids Res.1. 253-253 (1998)

[Publications] Sasaki,T.: "Specification of Initation Regions during Embryogenesis in the 65 kilobase DNA pol α-dE2F Locus of Drosophila melanogaster." Mol.Cel.Biol.,. 19,1. 547-555 (1999)

[Publications] Y.Ogawa,T: "Association of autonomous replication activity with replication origins in a human chromosome." Exp.Cell Res.243. 50-58 (1998)

[Publications] Horie,S.: "The Schizosaccharomyces pombe mei4+ gene encodes a meiosis-specific transcription factor containing a forkhead" Mol.Cell.Biol.18. 2118-2129 (1998)

[Publications] Tahara,S.: "Functional domains of Rep2,a transcriptional activator subunit for Res2-Cdc10,controlling the cell cycle“start"." Mol.Biol.Cell.9. 1577-1588 (1988)

[Publications] Shirahige,K: "Regulation of DNA replication origins during cell-cycle progression." Nature. 395. 618-621 (1998)

[Publications] Horie,T.: "A 5'-monophosphate form of bredinin selectively inhibits the activities of mammalian DNA polymerases in vitro." Intn.J.Mol.Med.1. 83-90 (1998)

[Publications] Ohta,K.: "Sulfoquinovosyldiacylglycerol,KM043,a new potent inhibitor of eukaryotic DNA polymerases and HIV-reverse transcriptase type I from" Gigartina tenella.Chem.Pharm.Bull.46. 684-686 (1998)

[Publications] Mizushina,Y.: "The inhibitory effect of novel triterpenoid compounds,fomitellic acids, on DNA polymerase b." Biochem.J.330. 1325-1332 (1998)

[Publications] Mizushina,Y.: "The biochemical inhibition mode of bredinin-5'-monophosphate on DNA polymerase b.Biochim." Biophys.Acta.1403. 5-11 (1998)

[Publications] Ohno,K.: "cDNA cloning and expression during development of Drosophila melanogaster MCM3,MCM6 and MCM7." Gene. 217. 177-186 (1998)

[Publications] Mizushina,Y.: "A mushroom fruiting body-inducing substance inhibits activities of replicative DNA polymerases." Biochem.Biophys.Res.Commun.249. 17-22 (1998)

[Publications] Ohtoshi,A: "Two recessive modes of growth inhibitionby exogenously introduced mutant genes: Analysis of mutant CDC28 and CDC7 genes" J.Biochem.Mol.Biol.Biophys.1. 253-263 (1998)

[Publications] Kim,J.M.: "Growth regulation ofthe expression of mouse cDNA and gene encoding a serine/threonine kinase related toS.cerevisiae CDC7 essential for" J.Biol.Chem.273. 23248-23257 (1998)

[Publications] Jonstone,L.: "A Cdc7-Dbf4 kinase activity is conserved from yeast to human" Progress in Cell Cycle Research. Vol.4(in press). (1999)

[Publications] 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)

[Publications] Mizushina,Y.: "Studies on inhibotors of mammalian DNA polymerase a and b. Sulfolipids from a pteridophyte,Athyrium niponicum." Biochem.Pharmacol.55. 537-541 (1998)

[Publications] Togashi,H.,: "4-Hydroxy-17-methylincisterol,and inhibitor of DNA polymerase-a activity and the growth of human cancer cell in vitro." Biochem.Pharmacol.56. 583-590 (1998)

[Publications] Tanaka,H.,: "Fission yeast Cdc24 is a replication factor C- and proliferating cell nuclear antigen-interacting factor essential for S phase completion." Mol.Cell.Biol.,. 19. 1038-1048 (1999)

[Publications] 片山 勉: "DNAポリメラーゼの新機能:染色体複製開始の負の制御" 蛋白質核酸酵素. (印刷中). (1999)

[Publications] 滝澤温彦: "MCM/P1ファミリー蛋白質とDNA複製のライセンス化" 実験医学. 16巻8号. 61-66 (1998)

[Publications] 白髭克彦: "複製開始領域の構成と染色体構築-出芽酵母の場合" 実験医学. 16. 31-40 (1998)

[Publications] 白髭克彦: "酵母染色体のマルチレプリコン構造とチェックポイント制御 細胞周期研究の新しい展開(田矢,野島編) (印刷中)" 羊土社,

滝澤温彦大阪大学, 大学院・理学研究科, 助教授 (60154944)

[Publications] 片山勉: "DNAポリメラーゼの新機能:染色体複製開始の負の制御" 蛋白質核酸酵素. (印刷中). (1999)

[Publications] 白髭克彦: "酵母染色体のマルチレプリコン構造とチェックポイント制御細胞周期研究の新しい展開(田矢,野島編) (印刷中)" 羊土社,