1992 Fiscal Year Annual Research Report

ゲノム複製開始の制御

Research Project

Project/Area Number	04256102
Research Institution	Osaka University
Principal Investigator	吉川寛大阪大学, 医学部, 教授 (70019876)
Co-Investigator(Kenkyū-buntansha)	山口和男金沢大学, 遺伝子実験施設, 教授 (00019879) 堀内嵩基礎生物学研究所, 教授 (60108644) 堀内賢介国立遺伝研, 細胞遺伝, 教授 (70219210) 今本文男京都薬科大, 生命薬学研, 教授 (00029761) 小笠原直毅大阪大学, 医学部, 講師 (10110553)
Keywords	ゲノム複製 / 複製開始点 / ARS配列 / 複製遺伝子 / 複製酵素
Research Abstract	1.原核生物の研究:正井らはプライミングシグナル上での精製蛋白によるプライマー合成反応の組識的な研究によって、大腸菌の多種類のprimosomeをその分子構成と識別配列の違いで分類することに成功した。小笠原は枯草菌のoriCプラスミドの分離に成功し、これを素材にして、複製開始のin vitro実験に成功した。これは大腸菌に次ぐ第二の系であり、開始の不偏性と多様性の分子機構の研究に進展が期待できる。榊原は大腸菌の新しく同定した複製開始遺伝子dnaR(PRPP合成酵素遺伝子)がDnaA蛋白と相互作用することを発見、ゲノムレベルの複製開始の調節に特異的に機能する可能性を示唆した。 2.真核生物の研究:吉川は出芽酵母第VI染色体の全ての自己複製配列(ARS)を分離同定、ゲノム上の位置を決定し、さらにすべてのARSについて、開始に必須なシス調節領域の構造を明かにした。升方はARSの存在すら殆ど分かっていないヒトのゲノムDNAから組織的なARSの分離に挑戦し、10-15kbの巨大なARSプラスミドの分離に成功している。杉野は酵母の複製酵素群の分離精製と機能の解析を組織的に行い、3種類のポリメラーゼの機能分担を明かにし、複製の″3ポリメラーゼ説″を提唱した。特に最近、トポイソメラーゼの分離精製に成功し、これを従来の酵素群に加えて、優れたin vitro複製系を初めて確立した。その他のin vitro複製では、SV40ウイルスゲノムをモデルとして、複製におけるクロマチン(ヌクレオソーム)の調節的機能の研究が進展した。 3.新しい方法の開発:真核生物のゲノムの複製開始点の同定を目標に2次元電気泳動方の改良、複製領域及び複製順序の決定に有効なFISH法の改良が行われ、班員間での共同研究の可能性が開けた。

Research Products
(15 results)

All Other

All Publications (15 results)

[Publications] Shirahige,K.: "Location and characterization of autonomously replicationg sequences(ARSs)from chromosome VI of Saccharomyces serecisiae" Mol.Cel.Biol. (1993)
[Publications] Ishimi,Y.: "Preincubation of T-antigen with DNA overcomes repression of SV40DNA replication by nucleosome assembly" J.Biol.Chem.267. 10910-10913 (1992)
[Publications] Ishimi,Y.: "Topoisomerase-II plays anessential role as a swivelase in the late stage of SV40 chromosome replication in vitro" J.Biol.Chem.267. 462-466 (1992)
[Publications] Sugasawa,K.: "Nonconservarive segregation of parental nucleosomes during simian virus-40 chromosome replication in vitro" Proc.Natl.Acad.Sci.USA. 89. 1055-1059 (1992)
[Publications] Yasuzawa,K.: "Histone-like proteins are required for cell growth and constraint of supercoils in DNA" Gene. 122. 9-15 (1992)
[Publications] Moriya,S.: "Cloning of autonomously replicating sequence(ars)fron the Bacillus subtilis chromosome" Mol.Microbiol.6. 309-315 (1992)
[Publications] Ogasawara,N.: "Genes and their organization in replication origin region of bacterial chromosome" Mol.Microbiol.6. 629-634 (1992)
[Publications] Fujita,M.Q.: "Structure of the DnaA and DnaA-box region in the Mycoplasma capricolum chromosome-Conservation and variations in the course of evolution" Gene. 110. 17-23 (1992)
[Publications] Araki,H.: "DNA polymerase II,the probable homolog of mammalian DNA polymerase ε, replicates chromosomal DNA in yeast SaccharomyCES cerecisiae" EMBO J.11. 733-740 (1992)
[Publications] Kitada,K.: "Temperature-sensitive cdc7 mutatuions of Saccharomyces cerevisiae are suppressed by the DBF4 Gene,which Is required for the G1/S cell cycle transition" Genetics. 131. 21-29 (1992)
[Publications] Higashitani,A.: "A single amino acid substitution reduces the superhelicity requirement of a replication initiator protein" Nucleic Acids Res.20. 2685-2691 (1992)
[Publications] Higashitani,N.: "SOS induction in Escherichia coli by infection with mutant filamentous phage that are defective in initiation of complementary-strand DNA synthesis" J.Bacteriol.174. 1612-1618 (1992)
[Publications] Hidaka,M.: "Termination complex in Escherichia coli inhibits SV40DNA replication in vitro by impeding the action of T-antigen helicase" J.Biol.Chem.267. 5361-5365 (1992)
[Publications] Kobayashi,T.: "Identification of a site required for DNA replication fork blocking activity in the rRNA gene cluster in Saccharomyces cerevisiae" Mol.Gen.Genet.233. 355-362 (1992)
[Publications] Maki,H.: "MutT protein specifically hydrolyzes 8-oxodGTP,a potent mutagenic substrate for DNA synthesis" Nature. 355. 273-275 (1992)

1992 Fiscal Year Annual Research Report

ゲノム複製開始の制御

Principal Investigator

吉川 寛 大阪大学, 医学部, 教授 (70019876)

Research Products

[Publications] Shirahige,K.: "Location and characterization of autonomously replicationg sequences(ARSs)from chromosome VI of Saccharomyces serecisiae" Mol.Cel.Biol. (1993)

[Publications] Ishimi,Y.: "Preincubation of T-antigen with DNA overcomes repression of SV40DNA replication by nucleosome assembly" J.Biol.Chem.267. 10910-10913 (1992)

[Publications] Ishimi,Y.: "Topoisomerase-II plays anessential role as a swivelase in the late stage of SV40 chromosome replication in vitro" J.Biol.Chem.267. 462-466 (1992)

[Publications] Sugasawa,K.: "Nonconservarive segregation of parental nucleosomes during simian virus-40 chromosome replication in vitro" Proc.Natl.Acad.Sci.USA. 89. 1055-1059 (1992)

[Publications] Yasuzawa,K.: "Histone-like proteins are required for cell growth and constraint of supercoils in DNA" Gene. 122. 9-15 (1992)

[Publications] Moriya,S.: "Cloning of autonomously replicating sequence(ars)fron the Bacillus subtilis chromosome" Mol.Microbiol.6. 309-315 (1992)

[Publications] Ogasawara,N.: "Genes and their organization in replication origin region of bacterial chromosome" Mol.Microbiol.6. 629-634 (1992)

[Publications] Fujita,M.Q.: "Structure of the DnaA and DnaA-box region in the Mycoplasma capricolum chromosome-Conservation and variations in the course of evolution" Gene. 110. 17-23 (1992)

[Publications] Araki,H.: "DNA polymerase II,the probable homolog of mammalian DNA polymerase ε, replicates chromosomal DNA in yeast SaccharomyCES cerecisiae" EMBO J.11. 733-740 (1992)

[Publications] Kitada,K.: "Temperature-sensitive cdc7 mutatuions of Saccharomyces cerevisiae are suppressed by the DBF4 Gene,which Is required for the G1/S cell cycle transition" Genetics. 131. 21-29 (1992)

[Publications] Higashitani,A.: "A single amino acid substitution reduces the superhelicity requirement of a replication initiator protein" Nucleic Acids Res.20. 2685-2691 (1992)

[Publications] Higashitani,N.: "SOS induction in Escherichia coli by infection with mutant filamentous phage that are defective in initiation of complementary-strand DNA synthesis" J.Bacteriol.174. 1612-1618 (1992)

[Publications] Hidaka,M.: "Termination complex in Escherichia coli inhibits SV40DNA replication in vitro by impeding the action of T-antigen helicase" J.Biol.Chem.267. 5361-5365 (1992)

[Publications] Kobayashi,T.: "Identification of a site required for DNA replication fork blocking activity in the rRNA gene cluster in Saccharomyces cerevisiae" Mol.Gen.Genet.233. 355-362 (1992)

[Publications] Maki,H.: "MutT protein specifically hydrolyzes 8-oxodGTP,a potent mutagenic substrate for DNA synthesis" Nature. 355. 273-275 (1992)

吉川寛大阪大学, 医学部, 教授 (70019876)