1991 Fiscal Year Annual Research Report

DNA複製反応の促進及び抑制のメカニズムと癌化

Research Project

Project/Area Number	03151025
Research Institution	Nagoya University
Principal Investigator	吉田松年名古屋大学, 医学部, 助教授 (70090420)
Co-Investigator(Kenkyū-buntansha)	有賀寛芳北海道大学, 薬学部, 教授 (20143505) 秋山徹大阪大学, 微生物病研究所, 助教授 (70150745) 小池克郎癌研究会癌研究所, 部長 (30085625) 安藤俊夫愛知県がんセンター, 研究所, 部長 (20012693) 花岡文雄理化学研究所, 主任研究員 (50012670)
Keywords	SV40ミニクロモソ-ム / DNAポリメラ-ゼ / プライマ-ゼ / DNAヘリカ-ゼ / トポイソメラ-ゼ / HBVXタンパク / RBタンパク / DNAヘリカ-ゼ
Research Abstract	ln vitro複製系:SV40ミニ染色体を鋳型としてDNAポリメラ-ゼα・プライマ-ゼ複合体、DNAポリメラ-ゼδ、PCNA等を用いた複製再構成系を確立し、これを用いてヌクレオソ-ムの複製時における再分配の様式を詳細に検討し、複製後leading及びlagging両鎖に平等に分配される事を示した(花岡)。新規トポイソメラ-ゼII阻害剤ICRM193はこの系の複製最終段階である両鎖のdecatenation及びTer領域の複製を阻害した(安藤)。複製開始及びlagging鎖上での不連続DNA合成に際してprimerRNAを合成するプライマ-ゼの塩基配列特異性に関しては従来不明であったが、牛胸腺DNAポリメラ-ゼα・プライマ-ゼ複合体が特に効率良く複製を開始する領域を解析してみると、その部位にはパリンドロ-ム構造及びTCTCTTA(有賀らの見出したcーmycARSのcore配列)を含み、哺乳類クロマチン複製開始起点との関連が注目される(吉田)。SV40DNA複製系に於て必須であるT抗原の宿主細胞側のカウンタ-パ-トの候補の一つとして、T抗原と同様にDNAポリメラ-ゼαと結合するDNAヘリカ-ゼを同定した。今後の解析が期待される。癌遺伝子産物及び癌抑制遺伝子産物:oncoprotein及びantioncoproteinと複製との関連につき進展がみられた。B型肝炎virusXタンパクを大腸菌で過剰発現、精製して生化学的特質を検討したところ、ATP、特異なタンパク分解酵素と結合能を有し、一方、転写調節因子とも結合し種々の因子の細胞内濃度の調節機能が示唆された(小池)。RBタンパクの細胞内標的として、RB結合タンパクを入gt11libraryでスクリ-ニングし結合タンパクをcodeするクロ-ン4種を得た。その一つ、56KDaタンパクを精製した。その生化学特質に興味が持たれる。RBタンパクはcdk2キナ-ゼでリン酸化修飾され、細胞増殖周期の進行を調節するが、その詳細な分子機構の解明が進行中である(秋山)。

Research Products
(14 results)

All Other

All Publications (14 results)

[Publications] Yoshida,S.Iwata,M.: "Sequence-dependent termination of mammlian DNA polymerase reaction by a new platinum compound (-)-(R)-2-aminomethylpyrrolidine(1.1-cyclobutane-dicarboxylato)platinum(II) monohydrate." Jpn.J.Cancer Res.82. 433-498 (1991)
[Publications] Izuta,S.: "The 5'ーtriphosphate of 3'ーazidoー3'ーdeoxythymidine and 2',3'ーdideoxynucleosides inhibit DNA polymerase γ by different mechanisms." Biochem.Biophys.Res.Commun.179. 776-783 (1991)
[Publications] Shioda,M.: "Stimulation of DNA polymerase α activity by microtubuleーassociated proteins." Biochemistry. 30. 11403-11412 (1991)
[Publications] Fumio Hanaoka Yukio Ishimi: "Replication of the simian virus 40 chromosome with purified proteins" J.Biol.Chem.266. 16141-16148 (1991)
[Publications] Kaoru Sugasawa: "Non-conservative segregation of parental nucleosomes during simian virus 40 chromosome replication in vitro" Proc.Natl.Acad.Sci.USA. in press.
[Publications] Anfoh,T.Ishimi,Y.: "Characterization of a camptothecin-resistant human DNA topoisomerase I in an in vitro system for simian virus 40 DNA replication." Eur.J.Biochem.
[Publications] Tsurumi,T.: "Characterization of 3'-to-5' exonuclease activity associated with Epstein-Barr virus DNA polymerase." Virology. 182. 376-381 (1991)
[Publications] Hirose,F.: "Structure and expression during development of Drosophila melanogaster gene for DNA polymerase." Nucl.Acids Res.19. 4991-4998 (1991)
[Publications] T.Akiyama: "The transforming potential of the c-erbB-2 protein is tegulated by its autophosphorylation at the carboxyl-terminal domain." Mol.Cell.Biol.11. 833-842 (1991)
[Publications] Ariga,H.Taira,T.: "c-myc protein complex binds to two sites of the human hsp 70 promoter region." Biochem.Biophys.Acta.
[Publications] Kitaura,H.: "Activation of c-myc promoter by c-myc protein in serum starved cells." FEBS Lett.290. 147-152 (1991)
[Publications] Katsuro Koike Masayuki Arii: "Identification of three essential regions of hepatitis B virus X protein for trans-activation function" Oncogene.
[Publications] Arai,K.Kitamura,T.: "Expression cloning of human IL-3 receptor cDNA and reconstitution of high affinity receptors for human IL-3 and GM-CSF" Cell. 66. 1165-1174 (1991)
[Publications] Abe,E.: "Eleven-base pair DNA sequence motif unique to human IL-4 gene confers responsiveness to T cell activation signals." Proc.Natl.Acad.Sci.USA.

1991 Fiscal Year Annual Research Report

DNA複製反応の促進及び抑制のメカニズムと癌化

Principal Investigator

吉田 松年 名古屋大学, 医学部, 助教授 (70090420)

Research Products

[Publications] Yoshida,S.Iwata,M.: "Sequence-dependent termination of mammlian DNA polymerase reaction by a new platinum compound (-)-(R)-2-aminomethylpyrrolidine(1.1-cyclobutane-dicarboxylato)platinum(II) monohydrate." Jpn.J.Cancer Res.82. 433-498 (1991)

[Publications] Izuta,S.: "The 5'ーtriphosphate of 3'ーazidoー3'ーdeoxythymidine and 2',3'ーdideoxynucleosides inhibit DNA polymerase γ by different mechanisms." Biochem.Biophys.Res.Commun.179. 776-783 (1991)

[Publications] Shioda,M.: "Stimulation of DNA polymerase α activity by microtubuleーassociated proteins." Biochemistry. 30. 11403-11412 (1991)

[Publications] Fumio Hanaoka Yukio Ishimi: "Replication of the simian virus 40 chromosome with purified proteins" J.Biol.Chem.266. 16141-16148 (1991)

[Publications] Kaoru Sugasawa: "Non-conservative segregation of parental nucleosomes during simian virus 40 chromosome replication in vitro" Proc.Natl.Acad.Sci.USA. in press.

[Publications] Anfoh,T.Ishimi,Y.: "Characterization of a camptothecin-resistant human DNA topoisomerase I in an in vitro system for simian virus 40 DNA replication." Eur.J.Biochem.

[Publications] Tsurumi,T.: "Characterization of 3'-to-5' exonuclease activity associated with Epstein-Barr virus DNA polymerase." Virology. 182. 376-381 (1991)

[Publications] Hirose,F.: "Structure and expression during development of Drosophila melanogaster gene for DNA polymerase." Nucl.Acids Res.19. 4991-4998 (1991)

[Publications] T.Akiyama: "The transforming potential of the c-erbB-2 protein is tegulated by its autophosphorylation at the carboxyl-terminal domain." Mol.Cell.Biol.11. 833-842 (1991)

[Publications] Ariga,H.Taira,T.: "c-myc protein complex binds to two sites of the human hsp 70 promoter region." Biochem.Biophys.Acta.

[Publications] Kitaura,H.: "Activation of c-myc promoter by c-myc protein in serum starved cells." FEBS Lett.290. 147-152 (1991)

[Publications] Katsuro Koike Masayuki Arii: "Identification of three essential regions of hepatitis B virus X protein for trans-activation function" Oncogene.

[Publications] Arai,K.Kitamura,T.: "Expression cloning of human IL-3 receptor cDNA and reconstitution of high affinity receptors for human IL-3 and GM-CSF" Cell. 66. 1165-1174 (1991)

[Publications] Abe,E.: "Eleven-base pair DNA sequence motif unique to human IL-4 gene confers responsiveness to T cell activation signals." Proc.Natl.Acad.Sci.USA.

吉田松年名古屋大学, 医学部, 助教授 (70090420)