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1994 Fiscal Year Final Research Report Summary

Mechanisms of molecular recognition and cell cycle control in ubiquitin-proteasome system

Research Project

Project/Area Number 05304054
Research Category

Grant-in-Aid for Co-operative Research (A)

Allocation TypeSingle-year Grants
Research Field Functional biochemistry
Research InstitutionHokkaido University, Faculty of Pharm.Sci.

Principal Investigator

YOKOSAWA Hideyoshi  Hokkaido Univ., Fac.pharm.sci., Prof., 薬学部, 教授 (90012765)

Co-Investigator(Kenkyū-buntansha) SAWADA Hitoshi  Hokkaido Univ., Fac.Pharm.Sci., Associate Prof., 薬学部, 助教授 (60158946)
ISHIKAWA Katsutoshi  Shizuoka Univ., Fac.Sci., Prof., 理学部, 教授 (00008608)
SAGATA Noriyuki  Kurume Univ.Inst.Life Sci., Prof., 分子生命科学研究所, 教授 (80142024)
SENO Takashi  Natl.Inst.Genet., Prof., 教授 (30076989)
SUZUKI Koichi  Tokyo Univ., Inst.Mol.Cell Biol., Prof., 分子細胞生物学研究所, 教授 (80011948)
Project Period (FY) 1993 – 1994
KeywordsProteasome / Ubiquitin / Protease / Molecular recognition / Cell cycle / Development / Fertilization / Oocyte maturation
Research Abstract

1.Physiological Substrates of Ubiquitin-conjugating Enzymes : (1) Ubiquitin-activating enzyme (E1)-defficient mutant cell (ts85) was found to undergo the abnormal features in nucleolus during cell cycle. (2) Proto-oncogene product, Mos, was found to be degraded by the 26S proteasome via ubiquitination of the dephosphorylated Mos at Ser3 on fertilization of the Xenopus eggs. (3) We succeeded to establish the several monoclonal antibodies specific to the multi-ubiquitin chains of the polyubiquitinated proteins. By using this antibody, studies on intracellular localization and quantification of the multi-ubiquitinated proteins were performed.
Structure and Function of Proteasomes : (1) From the gene-desruption studies, it was suggested that a proteasome subunit, Y13, is involved in the ATP-dependent proteolytic activity of the 26 S proteasome. (2) It was revealed that there are two isofoms in the 26 S proteasome complex : one is a complex made up by the 20 S proteasome and the regulatory s … More ubunit complex at a ratio of one, while the other is a complex a ratio of one two. (3) The 26 S proteasome was found to have a protein kinase activity closely coupled to ATP-dependent protease activity. (4) Ultrastructure of the 26 S proteasome was analyzed by EM and TM-AFM.(5) Substrate specificity of the 20 S proteasome was analyzed by using oxidized insulim B-chain as a substrate. (6) cDNA of the LMP7 subunit of the frog 26 S proteasome was cloned and its structural analysis was performed. (7) It was suggested that the 26 S(970kDa) proteasome is involved in the sperm penetration through the vitelline coat, while the 20 S proteasome is involved in the sperm binding to the vitelline coat.
3.Mechanisms of Cell Cycle Control : (1) ATP-dependent proteasome activity was transiently activated at a prophase and metaphase during the cell cycle of ascidian embryos. It was demonstrated that this activation is due to the interconversion between 20 S proteasome and 26 S proteasome, which is induced by intracellular calcium mobilization. (2) A membrane-bound from of the proteasome, witch is distinct from the cytosol proteasome in the subunit compositions, was newly identified. Less

  • Research Products

    (42 results)

All Other

All Publications (42 results)

  • [Publications] Kawahara,H.: "Intracellular calcium mobilization regulates the activity of 26 S proteasome during the metaphase-anaphase transition in the ascidian meiotic cell cycle.21GC01:Developmental Biology" 166. 623-633 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fujimuro,M.: "Production and characterization of monoclonal antibodies specific to multi-ubiquitin chains of polyubiquitinated proteins." FEBS Letters. 349. 173-180 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Furuno,N.: "Suppression of DNA replication via Mos function during meiotic divisions in Xenopus oocytes." EMBO Journal. 13. 2399-2410 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takahashi,M.: "EFP-sensitive multicatalytic protease complexes(proteasomes) involved in the control of oocyte maturation in the toad,Bufo japonicus." Molecular Reproduction and Development. 38. 310-317 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ohkubo,Y.: "SV40 large T antigen reinduces the cell cycle in terminally differentiated myotubes through inducing cdk2,cdc2,and their partner cyclins." Experimental Cell Research. 214. 270-278 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sudha,T.: "Abnormal integrity of the nucleolus associated with cell cycle arrested owing to the temperature-sensitive ubiquitin-activating enzyme E1." Chromosome Research. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saitoh,Y.: "High-molecular weight protease complexes (proteasomes) of sperm of the ascidian,Halocynthia roretzi:Isolation,characterization,and physiological roles in fertilization." Developmental Biology. 158. 238-244 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takizawa,S.: "Effects of protease inhibitors on binding of sperm to the vitelline coat of ascidian eggs:Implications for participation of a proteasome 〔multicatalytic proteinase complex〕." Journal of Experimental Zoology. 267. 86-91 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sawada,H.: "Different ratios in 20 S proteasomes and regulatory subunit complexes in two isoforms of the 26 S proteasome purified from rabbit skeletal muscle." FEBS Letters. 335. 207-212 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nishizawa,M.: "Degradation of Mos by the N-terminal proline (Pro^2)-dependent ubiquitin pathway on fertilization of Xenopus eggs:Possible significance of natural selection for Pro^2." EMBO Journal. 12. 4021-4027 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ishida,N.: "Mos is degraded by the 26 S proteasome in a ubiquitin-dependent fashion." FEBS Letters. 324. 345-348 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Honda,R.: "Dephosphorylation of human p34^<cdc2> kinase on both Thr-14 and Tyr-15 by human cdc25B phosphatase." FEBS Letters. 318. 331-334 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuda,H.: "The characterization of human cdc2 kinase and CDKs." Yakugaku Zasshi. 113. 829-846 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yamao,F.: "Conditional resistance to thymineless death predominantly selects DNA synthesis-deficient mutants of mammalian cells." Mutation Research. 289. 83-89 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takahashi,T.: "Cleavage specificity and inhibition profile of proteasome isolated from the cytosol of Xenopus oocyte." Journal of Biochemistry. 113. 225-228 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tokumoto,T.: "A novel ″active″form of proteasomes from Xenopus laevis ovary cytosol." Biochem.Biophys.Res.Commun.192. 1106-1114 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tokumoto,T.: "20 S latent proteasomes isolated from the cytosol of Xenopus oocytes:Purification and partial characterization." Biomedical Research. 14. 391-401 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Lee,D.H.: "Structure and properties of the 26 S protease complex from chick skeletal muscle." Biochem.Mol.Biol.Int.30. 212-130 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Fukushima,D.: "Activation of the secretory pathway leads to a decrease in the intracellular amyloidogenic fragments generated from the amyloid protein precursor." Biochem.Biophys.Res.Commun.194. 202-207 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saido,T.C.: "Spatial resolution of fodrin proteolysis in postischemic brain." Journal of Biological Chemistry. 268. 25239-25243 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saitoh, Y., Sawada.H., and Yokosawa, H.: "High-molecular-weight protease complexes(proteasomes)of sperm of the ascidian, Halocynthia roretzi : Isolation, characterization, and physiological roles in fertilization." Dev.Biol. 158. 238-244 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takizawa, S., Sawada, H., Someno, T., Saitoh, Y., Ykosawa, H., and Hoshi, M.: "Effects of protease inhibitors on binding of sperm to the vitelline coat of ascidian eggs : Implications for participation of a proteasome(multicatalytic proteinase complex)." J.Exp.Zool. 267. 86-91 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sawada, H., Muto, K., Fujimmuro, M., Akaishi, T., Takagi-Sawada, M., Yokosawa, H., and Goldberg, A.L.: "Different ratios in 20 S proteasomes and regulatory subunit complexes in two isoforms of the 26 proteasome purifies from rabbit skeletal muscle." FEBS Lett. 335. 207-212 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nishizawa, M., Furuno, N., Okazaki, K., Tanaka, H., Ogawa, Y., and Sagata, N.: "Degradation of Mos by the N-terminal proline(Pro^2)-dependent ubiquitin pathaway on fertilization of Xenopus eggs : possible signficance of natural selection for Pro^2." EMBO J.12. 4021-4027 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Ishida, N., Tanaka, K., Tamura, T., Nishizawa, M., Okazaki, K., Sagata, N., Ichihara, A.: "Mos is degraded by the 26 S proteasome in a ubiquitin-dependent fashion." FEBS Lett.324. 345-348 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Honda, R., Ohba, Y., Nagata, A., Okayama, H., and Yasuda, H.: "Dephosphorylation of human p34cdc2 kinase on both Thr-14 and Tyr-15 by human cdc25B phosphatase. FEBS Lett.318,331-334,1993." FEBS Lett.318. 331-334 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuda, H., Kamijo, M., and Ohba, Y.: "The characterization of human cdc2 kinase and CDK2" Yakugaku Zasshi. 113. 829-846 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yamao, F., Nagai, Y., Kaneda, S., Yoshida, S., and Seno, T.: "Conditional resistance to thymineless death predominantly selects DNA synthesis-deficient mutants of mammalian cells." Mutat.Res.289. 83-89 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, T., Tokumoto, T., Ishikawa, K., and Takahashi, K.: "Cleavage specificity and inhibition profile of proteasome isolated from the cytosol of Xenopus oocyte." J.Biochiem.113. 225-228 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tokumoto, T., and Ishikawa, K.: "A novel "active" form of proteasomes from Xenopus laevis ovary cytsol.Biochem.Biophys.Res.Commun." Biochem.Biophys.Res.Commun.192. 1106-1114 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tokumoto, T., and Ishikawa, K.: "20 S latent proteasomes isolated from the cytosol of Xenopus oocytes : Purificatin and partial characterization." Biomed.Res.14. 391-401 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Lee, D.H., Kim, S.S., Kim, K.I., Ahn, J.Y., Shim, K.S., Nishigai, M., Ikai, A,Tamura, T., Tanaka, K., Ichihara, A., Ha, D,B., and Chung, C.H.: "Structure and properties of the 26 S protease complex from chick skeletal muscle." Biochem.Mol.Biol.Int.30. 121-130 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fukushima, D., Konishi, M., Matuyama, K., Miyamoto, T., Ishiura, S., and Suzuki, K.: "Activation of the secretory pathwar leads to a decrease in the intracellular amyloidogenic fragments generated from the amyloid protein precursor." Biochem.Biophys.Res.Commun.194. 202-207 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Saido, T.C., Yokota, M., Nagao, S., Yamamura, I., Tani, E., Tsuchiya, T., Suzuki, K., and Kawashima, S.: "Spatial resolution of fodrin proteolysis in postischemic brain." J.Biol.Chem.268. 25239-25243 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kawahara, H., and Yokosawa, H.: "Intracellular calcium mobilization regulates the activity of 26 S proteasome during the metaphase-anaphase transition in the ascidian meiotic cell cycle." Dev.Biol.166. 623-633 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Fujimuro, M., Sawada, H., and Yokosawa, H.: "Production and characterization of monoclonal antibodies specific to multi-ubiquition chains of polyubiquitinated proteins.FEBS Lett." FEBS Lett.349. 173-180 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Furuno, N., Nishizawa, M., Okazaki, K., Tanaka, H., Iwashita, J., Nakajo, N., Ogawa, Y., and Sagata, N.: "Suppression of DNA replication via Mos function during meiotic divisions in Xenopus oocytes." EMBO J.13. 2399-2410 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahashi, M., Tokumoto, T., and Ishikawa, K.: "DEP-sensitive multicatalytic protease complexes(proteasomes)involved in the control of oocyte maturation in the toad, Bufo japonicus." Mol.Reprod.Dev.38. 310-317 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kato, Y., Salter-Cid, L., Flajnik, M.F., Kasahara, M., Namikawa, C,Sasaki, M., and Nonaka, M.: "Isolation of the Xenopus complement factor B complementary DNA and linkage of the gene to the frog MHC." J.Immunol.153. 4546-4554 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ohkubo, Y., Kishimoto, T., Nakata, T., Yasuda, H., and Endo, T.: "SV40 large T antigen reinduces the cell cycle in terminally differentiated myotubes through inducing cdk2, cdc2, and their partner cyclins." Exp.Cell Res.214. 270-278 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Honda, R., Tanaka, H., Ohba, Y., and Yasuda, H.: "Mouse p87weel kinase is regulated by M-phase specific phosphorylation" Chromosome Res.(in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sudha, T., Tsuji, H., Sameshima, M., Matsuda, Y., Kaneda, S., Nagai, Y., Yamao, F., and Seno, T.: "Abnormal integrity of the nucleolus associated with cell cycle arrest owing to the temperature-sensitive ubiquitin-activating enzyme E1." Chromosome Res.(in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1996-04-15  

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