1998 Fiscal Year Annual Research Report

細胞周期制御とDNA複製

Research Project

Project/Area Number	08265102
Research Institution	Nara Institute of Science and Technology
Principal Investigator	釣本敏樹奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (30163885)
Co-Investigator(Kenkyū-buntansha)	藤田雅俊愛知県がんセンター研究所, 研究員 (30270713) 大谷清東京医科歯科大, 疾患遺伝子実験センター, 助手 (30201974) 村上洋太京都大学, ウイルス研究所, 助教授 (20260622) 正井久雄東京大学, 医科学研究所, 助教授 (40229349) 加藤順也奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (00273839)
Keywords	ORC / PCNA / 複製開始点結合因子 / S期導入因子 / ライセンシング因子 / MCM / 蛋白質リン酸化 / E2F-1
Research Abstract	DNA複製およびその調節機構は細胞周期の進行制御と密接な連係を持ち、これに関与する因子は細胞のがん化、がん抑制に深く係わっている。そこでS期導入シグナル→複製開始の活性化機構(S期導入因子)→複製開始点複合体→複製装置という情報の流れに関わる因子の実体とその機能、各因子間の情報伝達のしくみの解明を行い以下の成果を得た。 1. 多くの複製因子の発現が転写因子E2Fの制御下にあることを明らかにした。またE2Fの発現誘導に伴い複製因子のクロマチン結合が誘導されることを明らかにした。 2. ライセンシング因子MCMが細胞周期依存的リン酸化を受け、これがMCMの機能発現と密接に関係していること、このリン酸化にcdc2およびCDC7キナーゼが関与していることを明らかにした。 3. 酵母の複製開始に係わるCDC7キナーゼのヒトホモログが、複製開始に必須の機能を持つことを明らかにした。 4. 複製開始因子ORCがチェックポイント機構に関与すること、核マトリクスに結合し、クロマチン構造維持機構とつながりがあることを示した。 5. S期誘導因子や複製因子と相互作用する新規因子を同定した。以上の結果、S期導入シグナルから複製装置に至る経路のつながりを浮かび上げることができた。さらにこの研究課題の成果を基にして、すでにより高度なネットワークの解析が進行中である。これらの進展により、G1期因子から複製因子までの正確な道筋を明らかにすることが期待される。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Oku, T: "Functional sites of human PCNA which interact with p21(Cip1/Waf1), DNA polymerase δ and replication factor C." Genes Cells. 3. 357-369 (1998)
[Publications] Tsurimoto, T: "PCNA, a multifunctional ring on DNA." Biochem.Biophys.Acta. 1443. 23-39 (1998)
[Publications] Fujita, M: "Cell cycle dependent topological changes of chromosomal replication origins in Saccharomy-ces cerevisiae" Genes Cells. 3. 737-749 (1998)
[Publications] Ohtani, K.: "Regulation of cell growth-dependent expression of mammalian CDC6 gene by the cell cycle transcription factor E2F." Oncogene. 17. 1777-1785 (1998)
[Publications] Ohtani, K.: "Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F." Oncogene. in press. (1999)
[Publications] Ohtani, K.: "Molecular mechanisms of promoter regulation of the gp34 gene that is trans-activated by an oncoprotein Tax of human T cell leukemia virus type I." J.Biol.Chem.273. 14119-14129 (1998)
[Publications] Urano, T: "p57 Kip2 is degraded through the proteasome in osteoblasts stimulated to proliferation by transforming growth factor β1." J.Biol.Chem.(in press). (1999)
[Publications] Akamatsu, E.: "Transcription factor E2F and cyclin E/cdk2 complex cooperate to induce chromosomal DNA replication in Xenopus oocytes." J.Biol.Chem.273. 16494-16500 (1998)
[Publications] Kurokawa, K.: "p19ARF prevents G1 cyclin-dependent kinase activation by interacting with MDM2 and activating p53 in mouse fibroblasts." Oncogene. (in press). (1999)
[Publications] Tomoda, K.: "Jab 1 targets the cyclin-dependent kinase inhibitor p27Kip 1 for degradation." J.Biol.Chem.(in press). (1999)
[Publications] Chen, L.-F.: "The capacity of polymavirus enhancer binding protein 2aB(AML1/Cbta2)to stimulate polyomavirus DNA replication is related to its affinity for the nuclear matrix." Mol.Cell.Biol. 18. 4165-4176 (1998)
[Publications] Sanchez, J.P.: "Isolation, characterization and molecular cloning of a protein(Abp2)that binds to a Schizosaccharomyces pombe origin of replication(ars3002)." Mol.Cell.Biol.18. 1670-1681 (1998)
[Publications] Tsurumi, T.: "Overexpression, purification, and Helix-destabilizing properties of Epstein-Barr virus single-stranded DNA-binding protein." J.Gen.Virol.79. 1257-1264 (1998)
[Publications] Hiraiwa, A.: "Specific distribution patterns of hCDC47 expression in cutaneous diseases" J.Cutan.Pathol.25. 285-290 (1998)
[Publications] Fujita, M.: "Cell cycle- and chromatin binding state-dependent phosphorylation of human MCM heterohexameric complexes: a role for cdc2 kinase." J.Biol.Chem.273. 17095-17101 (1998)
[Publications] Jonstone, L.: "A Cdc7-Dbf4 kinase activity is conserved from yeast to human." Progress in Cell Cycle Research. (in press). (1999)
[Publications] Kim, J.M.: "Growth regulation of the expression of mouse cDNA and gene encoding aserine/threonine ki-nase related to S.cerevisiae CDC7 essential for G1/S transition." J.Biol.Chem.273. 23248-23257 (1998)
[Publications] Ohtoshi, A.: "Two recessive modes of growth inhibition by exogenously introduced mutant genes: Analysis of mutant CDC28 and CDC7 genes in Saccharomyces cerevisiae." J.Biochem.Mol.Biol.Biophys.1. 253-263 (1998)

1998 Fiscal Year Annual Research Report

細胞周期制御とDNA複製

Principal Investigator

釣本 敏樹 奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (30163885)

Research Products

[Publications] Oku, T: "Functional sites of human PCNA which interact with p21(Cip1/Waf1), DNA polymerase δ and replication factor C." Genes Cells. 3. 357-369 (1998)

[Publications] Tsurimoto, T: "PCNA, a multifunctional ring on DNA." Biochem.Biophys.Acta. 1443. 23-39 (1998)

[Publications] Fujita, M: "Cell cycle dependent topological changes of chromosomal replication origins in Saccharomy-ces cerevisiae" Genes Cells. 3. 737-749 (1998)

[Publications] Ohtani, K.: "Regulation of cell growth-dependent expression of mammalian CDC6 gene by the cell cycle transcription factor E2F." Oncogene. 17. 1777-1785 (1998)

[Publications] Ohtani, K.: "Cell growth-regulated expression of mammalian MCM5 and MCM6 genes mediated by the transcription factor E2F." Oncogene. in press. (1999)

[Publications] Ohtani, K.: "Molecular mechanisms of promoter regulation of the gp34 gene that is trans-activated by an oncoprotein Tax of human T cell leukemia virus type I." J.Biol.Chem.273. 14119-14129 (1998)

[Publications] Urano, T: "p57 Kip2 is degraded through the proteasome in osteoblasts stimulated to proliferation by transforming growth factor β1." J.Biol.Chem.(in press). (1999)

[Publications] Akamatsu, E.: "Transcription factor E2F and cyclin E/cdk2 complex cooperate to induce chromosomal DNA replication in Xenopus oocytes." J.Biol.Chem.273. 16494-16500 (1998)

[Publications] Kurokawa, K.: "p19ARF prevents G1 cyclin-dependent kinase activation by interacting with MDM2 and activating p53 in mouse fibroblasts." Oncogene. (in press). (1999)

[Publications] Tomoda, K.: "Jab 1 targets the cyclin-dependent kinase inhibitor p27Kip 1 for degradation." J.Biol.Chem.(in press). (1999)

[Publications] Chen, L.-F.: "The capacity of polymavirus enhancer binding protein 2aB(AML1/Cbta2)to stimulate polyomavirus DNA replication is related to its affinity for the nuclear matrix." Mol.Cell.Biol. 18. 4165-4176 (1998)

[Publications] Sanchez, J.P.: "Isolation, characterization and molecular cloning of a protein(Abp2)that binds to a Schizosaccharomyces pombe origin of replication(ars3002)." Mol.Cell.Biol.18. 1670-1681 (1998)

[Publications] Tsurumi, T.: "Overexpression, purification, and Helix-destabilizing properties of Epstein-Barr virus single-stranded DNA-binding protein." J.Gen.Virol.79. 1257-1264 (1998)

[Publications] Hiraiwa, A.: "Specific distribution patterns of hCDC47 expression in cutaneous diseases" J.Cutan.Pathol.25. 285-290 (1998)

[Publications] Fujita, M.: "Cell cycle- and chromatin binding state-dependent phosphorylation of human MCM heterohexameric complexes: a role for cdc2 kinase." J.Biol.Chem.273. 17095-17101 (1998)

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

[Publications] Kim, J.M.: "Growth regulation of the expression of mouse cDNA and gene encoding aserine/threonine ki-nase related to S.cerevisiae CDC7 essential for G1/S transition." J.Biol.Chem.273. 23248-23257 (1998)

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

釣本敏樹奈良先端科学技術大学院大学, バイオサイエンス研究科, 助教授 (30163885)