1994 Fiscal Year Annual Research Report

ゲノム複製開始の制御

Research Project

Project/Area Number	04256102
Research Institution	Nara Institute of Science and Technology
Principal Investigator	吉川寛奈良先端科学技術大学院大学, バイオサイエンス研究科, 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) 杉野明雄大阪大学, 微生物病研究所, 教授 (90231737) 小笠原直毅奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (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:小笠原は枯草菌のdnaA変異株のsuppressor遺伝子の解析、プラスミドレプリコンによるdnaA及びoriC欠失を相補できる株の解析を行った。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蛋白に加えRNAポリメラーゼとも機能的に相互作用することを明らかにした。今本は大腸菌ヒストン様蛋白ファミリーがDNA局所高次構造の形成を介して、複製蛋白の機能発現を補佐していることを解析した。このように細菌染色体複製については開始制御蛋白ネットワークの解析が進んだ。山口はプラスミドpSC101のRep蛋白の分子形態の変化と複製開始における機能変化を明かにし、堀内は繊維状ファージf1の複製開始蛋白によるDNAの高次構造の逐次適変化を明らかにするなど、複製開始蛋白の詳細な解析を発展させた。 2.真核生物の研究:吉川は出芽酵母第VI染色体の自己複製配列(ARS)に対する、各種複製蛋白の変異の影響を系統的に解析し、シスとトランスの因子の相互作用の特異性を明らかにした。升方はヒトのゲノムDNAから自立複製能をエンハーンスする特異的配列を単離し、その構造を明らかにする共に、それが転写抑制機能を持つことを示した。四宮はショウジョウバエのDNA polymerase α遺伝子の下流に複製開始領域を同定し、その近傍21Kbの塩基配列を解析し、複製開始領域の両側には核骨格結合部位が存在することを明らかにした。杉野は複製の"3ポリメラーゼ説"を提唱したが、DNA polymeraseIIと相互作用する遺伝子産物を探索し、新たな必須遺伝子を同定し、それが複製のみならず細胞周期調節にも関与することを明らかにした。正井は出芽酵母のARSと相互作用する蛋白の単離を試み、いくつかの新たな蛋白遺伝子を同定した。このように、真核生物の複製研究についてもその多様な制御因子の解明が進んだ。

Research Products

(17 results)

All Other

All Publications (17 results)

[Publications] Yamazoe,M.: "A Protein which binds preferentially to single-stranded core sequence of autonomously replicating sequence is essential for respiratory function in mitochondria of S.cerevisiae." J Biol Chem. 269. 15244-15252 (1994)
[Publications] Moriya,S.: "Replication of a Bacillus subtilis oriC plasmid in vitro." Mol Microbiol. 12. 469-478 (1994)
[Publications] Rashid,M.B.: "Anatomy of the stimulative sequences flanking the ARS consensus of chromosome VI in Saccharomyces cerevisiae" Gene. 150. 213-320 (1994)
[Publications] Ishimi,Y.: "Loading of a DNA helicase on the DNA unwinding element in the yeast replication origin-mechanizm of DNA replication in a model system." Biochemistry. 33. 2733-2740 (1994)
[Publications] Ishimi,Y.: "DNA replication from initiation zones of mammalian cells in a model system." Mol Cell Biol. 14. 6489-6496 (1994)
[Publications] Wada,M.: "Increased retention of small chromosomes in a novel yeast mutant." J Biochem Tokyo. 116. 694-696 (1994)
[Publications] Sakakibara,Y.: "Suppression of thermosensitive initiation of DNA replication in a dnaR mutant of EScherichia coli by a rifamin resistance mutation in the rpoB gene." J Bacteriol. 177. 733-737 (1995)
[Publications] Shinomiya,T.: "Mapping an initiation region of DNA replication at a single-copy chromosomal locus in Drosophila melanogaster cells by two-dimensional gel methods." Mol Cell Biol. 14. 7394-7403 (1994)
[Publications] Leem,S.H.: "The yeast SaccharomycescerevisiaeDNA polymerase IV:possible involvement in double strand break DNA repair." Nucleic Acids Res. 22. 3011-3017 (1994)
[Publications] Morrison,A.: "The 3'->5' exonucleases of both DNA polymerases delta and epsilon participate in correcting errors of DNA replication in Saccharomycescerevisiae" Mol Gen Genet. 242. 289-296 (1994)
[Publications] Noskov,V.: "The RFC2 gene encoding a subunit of replication factor of Saccharomycescerevisiae" Nucleic Acids Res. 22. 1527-1535 (1994)
[Publications] Pack,R.A.: "Isolation of human DNA-unwinding elements as sites of DNA polymerase alpha/primase entry." Gene. 148. 277-284 (1994)
[Publications] Higashitani,A.: "Multiple DNA conformational changes induced by an initiator protein precede the nicking reaction in a rolling circle replication origin." J Mol Biol. 237. 388-400 (1994)
[Publications] Higashitani,A.: "A general and fast method for mapping mutations on the Escherichiacoli chromosome." Nucleic Acids Res. 22. 2426-2427 (1994)
[Publications] Masai,H.: "Escherichiacoli PriA protein is essential for inducible and constitutive stable DNA replication." EMBOJ. 13. 5338-5345 (1994)
[Publications] Ohkubo,S.: "Two enhancer elements for DNA replication of pSC101,par anda palindromic binding sequence of the rep protein." J Bacteriol. 177. 558-565 (1995)
[Publications] Suzuki,M.: "DNA polymerase alpha overcomes an error-prone pause site in the presence of replication protein-a." J Biol Chem. 269. 10225-10228 (1994)

1994 Fiscal Year Annual Research Report

ゲノム複製開始の制御

Principal Investigator

吉川 寛 奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (70019876)

Research Products

[Publications] Yamazoe,M.: "A Protein which binds preferentially to single-stranded core sequence of autonomously replicating sequence is essential for respiratory function in mitochondria of S.cerevisiae." J Biol Chem. 269. 15244-15252 (1994)

[Publications] Moriya,S.: "Replication of a Bacillus subtilis oriC plasmid in vitro." Mol Microbiol. 12. 469-478 (1994)

[Publications] Rashid,M.B.: "Anatomy of the stimulative sequences flanking the ARS consensus of chromosome VI in Saccharomyces cerevisiae" Gene. 150. 213-320 (1994)

[Publications] Ishimi,Y.: "Loading of a DNA helicase on the DNA unwinding element in the yeast replication origin-mechanizm of DNA replication in a model system." Biochemistry. 33. 2733-2740 (1994)

[Publications] Ishimi,Y.: "DNA replication from initiation zones of mammalian cells in a model system." Mol Cell Biol. 14. 6489-6496 (1994)

[Publications] Wada,M.: "Increased retention of small chromosomes in a novel yeast mutant." J Biochem Tokyo. 116. 694-696 (1994)

[Publications] Sakakibara,Y.: "Suppression of thermosensitive initiation of DNA replication in a dnaR mutant of EScherichia coli by a rifamin resistance mutation in the rpoB gene." J Bacteriol. 177. 733-737 (1995)

[Publications] Shinomiya,T.: "Mapping an initiation region of DNA replication at a single-copy chromosomal locus in Drosophila melanogaster cells by two-dimensional gel methods." Mol Cell Biol. 14. 7394-7403 (1994)

[Publications] Leem,S.H.: "The yeast SaccharomycescerevisiaeDNA polymerase IV:possible involvement in double strand break DNA repair." Nucleic Acids Res. 22. 3011-3017 (1994)

[Publications] Morrison,A.: "The 3'->5' exonucleases of both DNA polymerases delta and epsilon participate in correcting errors of DNA replication in Saccharomycescerevisiae" Mol Gen Genet. 242. 289-296 (1994)

[Publications] Noskov,V.: "The RFC2 gene encoding a subunit of replication factor of Saccharomycescerevisiae" Nucleic Acids Res. 22. 1527-1535 (1994)

[Publications] Pack,R.A.: "Isolation of human DNA-unwinding elements as sites of DNA polymerase alpha/primase entry." Gene. 148. 277-284 (1994)

[Publications] Higashitani,A.: "Multiple DNA conformational changes induced by an initiator protein precede the nicking reaction in a rolling circle replication origin." J Mol Biol. 237. 388-400 (1994)

[Publications] Higashitani,A.: "A general and fast method for mapping mutations on the Escherichiacoli chromosome." Nucleic Acids Res. 22. 2426-2427 (1994)

[Publications] Masai,H.: "Escherichiacoli PriA protein is essential for inducible and constitutive stable DNA replication." EMBOJ. 13. 5338-5345 (1994)

[Publications] Ohkubo,S.: "Two enhancer elements for DNA replication of pSC101,par anda palindromic binding sequence of the rep protein." J Bacteriol. 177. 558-565 (1995)

[Publications] Suzuki,M.: "DNA polymerase alpha overcomes an error-prone pause site in the presence of replication protein-a." J Biol Chem. 269. 10225-10228 (1994)

吉川寛奈良先端科学技術大学院大学, バイオサイエンス研究科, 教授 (70019876)