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Studies on the mechanism of cell division in animal cells.

Research Project

Project/Area Number 60065005
Research Category

Grant-in-Aid for Specially Promoted Research

Allocation TypeSingle-year Grants
Research InstitutionThe University of Tokyo

Principal Investigator

SAKAI Hikoichi  Dept. Biophys. Biochem., Fac. Sci. Univ. Tokyo Professor, 理学部, 教授 (80011477)

Co-Investigator(Kenkyū-buntansha) ENDO Sachiko  Dept. Biophys. Biochem., Fac. Sci. Univ. Tokyo Instructor, 理学部, 教務職員
MAEKAWA Shohei  Dept. Biophys. Biochem., Fac. Sci. Univ. Tokyo Assistant Professor, 理学部, 助手 (40173695)
NISHIDA Eisuke  Dept. Biophys. Biochem., Fac. Sci. Univ. Tokyo Assistant Professor, 理学部, 助手 (60143369)
MUROFUSHI Hiromu  Dept. Biophys. Biochem., Fac. Sci. Univ. Tokyo Associate Professor, 理学部, 助教授 (70101128)
Project Period (FY) 1985 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥185,000,000 (Direct Cost: ¥185,000,000)
Fiscal Year 1989: ¥10,000,000 (Direct Cost: ¥10,000,000)
Fiscal Year 1988: ¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 1987: ¥20,000,000 (Direct Cost: ¥20,000,000)
Fiscal Year 1986: ¥50,000,000 (Direct Cost: ¥50,000,000)
Fiscal Year 1985: ¥90,000,000 (Direct Cost: ¥90,000,000)
KeywordsGrowth factor / Transduction of growth signal / Protein kinase / MAP-2 kinase / Centrosome / Microtubules / Mitotic apparatu / Cleavage signal / プロテインキナーゼ / 微小管 / 細胞分裂 / DNA合成 / 輸送モーター / 輸送モータ / 蛋白リン酸化 / 有系分裂 / 分裂収縮環 / 微小管形成中心 / 染色体運動 / 増殖因子受容体 / 細胞骨格
Research Abstract

[Mechanism of signal transduction for call proliferatrion] : We demonstrated phosphorylation of high molecular weight microtubule-associated proteins (MAPS) as well as activation of a MAP-2 kinase after stimulation of cultured mammalian cells by growth factor or phorbol ester (promoter of carcinogenesis) and further identified a protein kinase which does not phosphorylate histone or casein but only MAP-2 and myelin basic-protein, proved to be a protein kinase newly identified. We further demonstrated that disassembly of microtubules produced by microtubule-disrupting drugs directly signals cell proliferation without stimulation by growth factor or pharbol ester. These results strongly suggest that MAP-2 kinase is involved in the cascade of phosphorylation in signal transduction for cell proliferation and demonstrate that microtubule cytoskeleton is directly relevant to the signal transduction, raising a now concept in the mechanism.
[Mechanism of the formation of the mitotic apparatus] … More : We identified the major protein component of the centrasons, 51-kD protein, and demonstrated its aster forming ability in the presence of some minor components of the contrasome and tubulin. The protein, which has an isoelectric point of 9.8 as basic protein, has an ability to bind guanine nucleotide, GTP and GDP, and proved to be a G protein. Furthermore, we demonstrated interconversion between GTP and GDP bound to the 51-kD protein, and the GTP-form protein is competent to be a signal protein for microtubule assembly. This provided a new standpoint in the mechanism of the formation of the mitotic apparatus, that is, the GTP<double arrow>GDP interconversion in the 51-kD protein sites governs the ability of the contrasome to initiate microtubules to build up the aster and the spindle. In fact, we demonstrated that isolated centrosomal fragments saturated with GTP (with the 51-kD protein saturated with GTP) can initiate astral microtubules much more efficiently than those saturated with GDP.
[Mechanism of transduction of cleavage signal] : We purified kinesin, which is most likely concerned with the trainduction of cleavage signal as a translocation motor, and characterized its enzymatic properties. We further isolated and purified a 260-kD protein which localizes underneath the cell membrane Just co-localized with actin bundles in the contractile ring. The 260-kD protein was shown to form curled thick actin bundles in vitro. Furthermore, an important finding is that a transmembrane glycoprotein labeled with WGA assembles as a precursor structure along the predicted contractile ring through the action of the cleavage signal which was shown to be dependent on microtubules, possibly astral microtubules. Less

Report

(6 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • 1985 Annual Research Report
  • Research Products

    (69 results)

All Other

All Publications (69 results)

  • [Publications] Miyata,Y.,Nishida,E.,Koyasu,S.,Yahara,I. & Sakai,H.: "Protein Kinase C-dependent and -independent pathways in the growth factor-induced cytoskeletal reorganization." J.Biol.Chem.264. 15565-15568 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hoshi,M.,Nishida,E. & Sakai,H.: "Characterizaation of a mitogen-activated,Ca^<2+>-sensitive microtubule-associated protein-2 kinase." Eur.J.Biochem.184. 447-486 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hoshi,M.,Nishida,E.,Matsumoto,S.,Akiyama,T.,Kawakami,M.,Yahara,I. & Sakai,H.: "Glucose and nitrogen rapidly activate a Ca^<2+>-inhibitable,serine/threonine kinase activity toward microtubule-associated protein 2 in Saccharomyces cerevisiae." FEBS Lett.258. 67-70 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Shinohara,Y.,Nishida,E. & Sakai,H.: "Initiation of DNA synthesis by microtubule disruption in quiescent rat 3Y1 cells." Eur.J.Biochem.183. 275-280 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Shioda,M.,Murofushi,H.,Murakami-Murofushi,K. & Sakai,H.: "Microtubule-associated protein-2 stimulates DNA synthesis catalyzed by the nuclrar matrix." Biochem.Biophys.Res.Commun.159. 834-841 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hamaguchi,Y.,Iwasa,F.,Toriyama,M. & Sakai,H.: "A comparative study of the distribution of fluorescently labeled calmodulin and tubulin in the meiotic apparatus of the mouse oocyte." Cell Struct.Funct.14. 241-248 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Okuhara,K.,Murofushi,H.& Sakai,H.: "Binding of Kinesin to stress fibers in fibroblasts under condition of microtubule depolymerization." Cell Motil.Cytoskel.12. 71-77 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshigaki,T.,Maekawa,S.,Endo,S.& Sakai,H.: "Localization of a high-molecular-weight actin binding protein in the sea urchin egg from fertilization through cleavage." Cell Struct.Funct.14. 363-374 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Aizawa, H., Kawasaki, H., Murofushi, H., Kotani, S., Suzuki, K. & Sakai, H.: ""A common amino acid sequence in 190-kDa microtubule-associated protein and tau for the promotion of microtubule-assembly"" J. Biol. Chem.264. 5885-5890 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Miyata, Y., Nishida, E., Koyasu, S., Yahara, I. & Sakai, H.: ""Regulation by intracellular Ca^<2+> and cyclic AMP of the growth factor-induced ruffling membrane formation and stimulation of fluid-phase endocytosis and exocytosis"" Exptl. Cell Res.181. 4544-4462 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Miyata, Y., Nishida, E., Koyasu, S., Yahara, I. & Sakai, H.: ""Protein kinase C-dependent and -independent pathways in the growth factor-induced cytoskeletal reorganization"" J. Biol. Chem.264. 15565-15568 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hoshi, M., Nishida, E. & Sakai, H.: ""Characterization of a mitogen-activated, Ca^<2+> -sensitive microtubule-associated" Eur.J. Biochem.184. 477-486 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hoshi, M., Nishida, E., Matsumoto, S., Akiyama, T., Kawakami, M., Yahara, I. & Sakai, H.: ""Glucose and nitrogen rapidly activate a Ca^<2+> -inhibitable, serine/threonine kinase activity toward microtubule-associated protein 2 in Saccgarintces cerevusuae." FEBS Lett.258. 67-70 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Shinohara, Y., Nishida, E. & Sakai, H.: ""Initiation of DNA synthesis by microtubule disruption in quiescent rat 3Y1 cells"" Eur. J. Biochem.183. 275-280 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Shioda, M., Murofushi, H., Murakami-Murofushi, K. & Sakai, H.: ""Microtubule-associated protein-2 stimulates DNA synthesis catalyzed by the nuclear matrix"" Biochem. Biophys. Res. Commun.159. 834-841 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hamaguchi, Y., Iwasa, F., Toriyama, M. & Sakai, H.: ""A comparative study of the distribution of fluorescently labeled calmodulin and tubulin in the meiotic apparatus of the mouse oocyte"" Cell Struct. Funct.14. 241-248 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Okuhara, K., Murofushi, H. & Sakai, H.: ""Binding of kinesin to stress fibers in fibroblasts under condition of microtubule depolymerization"" Cell Motil Cytoskel.12. 71-77 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yoshigaki, T., Maekawa, S., Endo, S. & Sakai, H.: ""Localization of a high-molecular-weight actin binding protein in the sea urchin egg from fertilization through cleavage"" Cell Struct. Funct.14. 363-374 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Miyata,Y.,Nishida,E.,Koyasu,S.,Yahara,I.& Sakai,H.: "Protein kinase C-dependent and -independent pathways in the growth factor-induced cytoskeletal reorganization." J.Biol.Chem.264. 15565-15568 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Hoshi,M.,Nishida,E.& Sakai,H.: "Characterizaation of a mitogen-activated,Ca^<2+>-sensitive microtubuleーassociated protein-2 kinase." Eur.J.Biochem.184. 477-486 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Hoshi,M.,Nishida,E.,Matsumoto,S.,Akiyama,T.,Kawakami,M.,Yahara,I.& Sakai,H.: "Glucose and nitrogen rapidly activate a Ca^<2+>-inhibitable,serine/threonine kinase activity toward microtubuleーassociated protein 2 in Saccharomyces cerevisiae." FEBS Lett.258. 67-70 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Shinohara,Y.,Nishida,E.& Sakai,H.: "Initiation of DNA synthesis by microtubule disruption in quiescent rat 3Y1 cells." Eur.J.Biochem.183. 275-280 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Shioda,M.,Murofushi,H.,Murakami-Murofushi,K.& Sakai,H.: "Microtubule-associated protein-2 stimulates DNA synthesis catalyzed by the nuclear matrix." Biochem.Biophys.Res.Commun.159. 834-841 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Hamaguchi,Y.,Iwasa,F.,Toriyama,M.& Sakai,H.: "A comparative study of the distribution of fluorescently labeled calmodulin and tubulin in the meiotic apparatus of the mouse oocyte." Cell Struct.Funct.14. 241-248 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Okuhara,K.,Murofushi,H&Sakai,H.: "Binding of kinesin to stress fibers in fibroblasts under condition of microtubule depolymerization." Cell Motil.Cytoskel.12. 71-77 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yoshigaki,T.,Maekawa,S.,Endo,S.& Sakai,H.: "Localization of a high-molecular-weight actin binding protein in the sea urchin egg from fertilization through cleavage." Cell Struct.Funct.14. 363-374 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Minako,Hoshi;Eisuke,Nishida;Hikoichi,Sakai: The Journal of Biological Chemistry. 263. 5396-5401 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Minako,Hoshi;Tetsu,Akiyama;Yukiko,Shinohara;Yoshihiko Miyata;Hiroshi,Ogawara;Eisuke,Nishida;Hikoichi,Sakai: European Journal of Biochemistry. 174. 225-230 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Eisuke,Nishida;Kazuyuki,Tobe;Takashi,Kadowaki;Masato,Kasuga;Chikako Sato;Hikoichi Sakai: Cell Structure and Function. 13. 417-423 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Yukiko,Shinohara;Eisuke,Nishida;Hikoichi,Sakai: FEBS LETTERS. 236. 19-22 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Kunihiro,Ohta;Masaru,Toriyama;Sachiko,Endo;Hikoichi,Sakai: Zoological Science. 5. 613-621 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Masarau,Toriyama;Kunihiro,Ohta;Sachiko,Endo;Hikoichi,Sakai: Cell Motility and the Cytoskeleton. 9. 117-128 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Kunihiro,Ohta;Masaru,Toriyama;Sachiko,Endo;Hikoichi,Sakai: Cell Motility and the Cytoskeleton. 10. 496-505 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Hiromu,Murofushi;Atsushi,Ikai;Koji,Okuhara;Susumu,Kotani;Hiroyuki,Aizawa;Konosuke,Kumakura;Hikoichi,Sakai: The Journal of Biological Chemistry. 263. 12744-12750 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Kadowaki,S.Koyasu,E.Nishida,K.Tobe,T.Izumi,F.Takaku,H.Sakai,I.Yahara, & M. Kasuga: The Journal of Biological Chemistry. 262. 7342-7350 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Akiyama,E.Nishida,J.Ishida,N.Saji,H.Ogawara,M.Hoshi, Y.Miyata, & H. Sakai: The Journal of Biological Chemistry. 261. 15648-15651 (1986)

    • Related Report
      1987 Annual Research Report
  • [Publications] Eisuke Nishida, Kazuko Iida, Naoto Yonezawa, Shigeo Koyasu, Ichiro Yahara, & Hikoichi Sakai: Proc. Natl. Acad. Sci. USA. 84. 5262-5266 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] M.Hoshi, E.Nishida, Y.Miyata, H.Sakai, T.Miyoshi, H.Ogawara, & T.Akiyama: FEBS Lett. (Federation of European Biochemical Societies). 217. 237-241 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] T.Kadowaki,M.Kasuga,K.Tobe,F. Takaku,E.Nishida,H.Sakai,S. Koyasu,I.Yahara,K.Toyoshima,T. Yamamoto,T.Akiyama: Biochemical and Biophysical Research Communications. 144. 699-704 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yasutaka Ohta, Tetsu Akiyama, Eisuke Nishida, & Hikoichi Sakai: FEBS Lett. (Federation of European Biochemical Societies). 222. 305-310 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Shin-ichi Hisanaga, Takeshi Tanaka, Tomoh Masaki, Hikoichi Sakai, Issei Mabuchi, & Yukio Hiramoto: Cell Motility and the Cytoskeleton. 7. 97-109 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yukihisa Hamaguchi,Masaru Toriyama,Hikoichi Sakai, & Yukio Hiramoto: Cell.Structure and Function. 12. 43-52 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Susumu Kotani, Hiromu Murofushi, Shohei Maekawa, Hiroyuki Aizawa, Kazuhiko. Kaji, & Hikoichi Sakai: Cell Struction and Function. 12. 1-9 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Hiroyuki Aizawa, Hiromu Murofushi, Susumu Kotani, Shin-ichi Hisanaga, Nobutaka Hirokawa, & Hikoichi Sakai: The Journal of Biological Chemistry. 222. 3782-3787 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] M.Toriyama,K.Ohta,S.Endo, & H.Sakai: Cell Motility and the Cytoskeleton. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Kunihiro Ohta, Masaru Toriyama, Yukihisa Hamaguchi, Shohei Maekawa, Sachiko Endo, & Hikoichi Sakai: Protoplasma. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Kunihiro Ohta, Masaru Toriyama, Sachiko Endo, & Hikoichi Sakai: Cell Motility and the Cytoskeleton. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Susumu Kotani, Hiromu Murofushi, Shohei Maekawa, Hiroyuki Aizawa, & Hikoichi Sakai: Journal of Biological Chemistry. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Akiyama,T.;Kadowaki,T.;Nishida,E.;Kadooka,T.;Ogawara,H.;Fukami,Y.;Sakai,H.;Takaku,F.;Kasuga,MM.: Journal of Biological Chemistry. 261. 314797-1480 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Kadowaki;T.,Koyasu;S.,Nishida,E.;Sakai,H.;Takaku,F.;Yahara,I.;Kosuga,M.: Journal of Biological Chemistry. 261. 716141-1614 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Akiyama,T.;Nishida,E.;Ishida,J.;Saji,N.;Ogawara,H.;Hoshi,M.;Miyata,Y.;Sakai,H.: Journal of Biological Chemistry. 261. 15648-15651 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Hosoya,H.;Iwasa,F.;Ohnuma,I.;Mabuchi,I.;Mohri,H.;Sakai,H.: FEBS Letters. 205. 121-126 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Hisanaga,S.;Tanaka,T.;Masaki,T.;Sakai,H.;Mabuchi,I.;Hiramoto,Y.: Cell Motility and Cytoskeleton. (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Hamaguchi,Y.;Toriyama,M.;Sakai,H.;Hiramoto,Y.: Cell Structure and Function. (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Kotani,S.;Murofushi,H.;Maekawa,s.;Sato,S.;Sakai,H.: European Journal of Biochemistry. 156. 23-29 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Murofushi,H.;Kotani,S.;Aizawa,H.;Hisanaga,S.;Hirokawa,M.;Sakai,H.: Journal of Cell Biology. 103. 1911-1919 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Kotani,S.;Murofushi,H.;Maekawa,S.;Aizawa,H.;Kaji,K.;Sakai,H.: Cell Structure and Function.

    • Related Report
      1986 Annual Research Report
  • [Publications] Kumagai,H.;Nishida,E.;Kotani,S.;Sakai,H.: Journal of Biochemistry(Tokyo). 99. 521-525 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Miyata,T.;Hoshi,M.;Nishida,E.;Minami,Y.;Sakai,H.: Journal of Biological Chemistry. 261. 013026-1303 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Nishida,E.;Koyasu,S.;Sakai,H.;Yahara,I.: Journal of Biological Chemistry. 261. 616033-1603 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Koyasu,S.;Nishida,E.;Kadowaki,T.;Matsuzaki,F.;Iida,K.;Harada,F.;Kasuga,M.;Sakai,H.;Yahara,I: Proceedings of National Acedemy of Science USA. 83. 8054-8058 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Toriyama,M.;Ohta,K.;Endo,S.;Sakai,H.: Cell Motility and Cytoskeleton.

    • Related Report
      1986 Annual Research Report
  • [Publications] Maekawa,S.;Endo,S.;Sakai,H.: Experimental Cell Research.

    • Related Report
      1986 Annual Research Report
  • [Publications] Endo,S.;Toriyama,M.;Sakai,H.: "Microtubule-organizing granules(MTOG)in mitotic sea urchin egg.In "CELL MOTILITY:MECHANISM AND REGULATION"H.Ishidawa,S.Hatano and H.Sato,editors pp.403-414" University of Tokyo Press, 627 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Biochem.Biophys.Res.Commun.127-2. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] J.Biol.Chem.260-19. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] J.Cell Biol.100-4. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] FEBS Lett.185-2. (1985)

    • Related Report
      1985 Annual Research Report
  • [Publications] Cell Struct.d Funct.10. (1985)

    • Related Report
      1985 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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