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1998 Fiscal Year Final Research Report Summary

Role of myosin-binding poteins in myofibrillogenesis

Research Project

Project/Area Number 08044191
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research Field 動物生理・代謝
Research InstitutionDEPARTMENT OF BIOLOGY,CHIBA UNIVERSITY

Principal Investigator

OBINATA Takashi  Chiba Univ.Dept.of Biology Professor, 理学部, 教授 (40012413)

Co-Investigator(Kenkyū-buntansha) STOCKDALE Fr  スタンフォード大学, 医学部, 教授
WELIKSON Rob  コーネル大学, 医学部, 研究員
FISCHMAN Don  コーネル大学, 医学部, 教授
SATO Naruki  Chiba Univ.Dept.of Biology Research Associate, 理学部, 助手 (40261896)
WELIKSON Robert E.  Cornell Univ. Medical School Research Associate
STOCKDALE Frank E.  Stanford Univ.Medical School Professor
FISCHMAN Donald A.  Cornell Univ Medical School Professor
Project Period (FY) 1996 – 1998
KeywordsC-protein / H-protein / Myosin-binding protein / Myofibril / Myofibrillogenesis / Muscle cells / Myosin / Actin
Research Abstract

C-protein is a major myosin-binding protein characteristic of striated muscles. In order to clarity the roles of C-protein in myofibrillogenesis, we carried out the following experiments and obtained the results as described.
1) Mouse C-protein cDNA sequences and C-protein isoform expression
The cDNAs encoding mouse fast skeletal (F) and slow skeletal (S) C-proteins were cloned and their entire sequences were determined. Cardiac (C)-isoform is known to be expressed first and transiently during development of chicken skeletal muscles, but in mouse skeletal muscle, C-isoform was not detected through developmental stages. S-isoform became detectable first, followed by appearance of F-isoform, In the degenerating mouse skeletal muscles, expression of S-isoform is increased.
2) Two isoforms in chicken cardiac C-protein and their expression
We detected two variants (type I and type II) in chicken cardiac C-protein and determined their entire sequences ; they differ only by the inclusion or exclu … More sion of a sequence of 15 amino acid residues (P-seq) which includes a phosphorylation site for A-kinase. We prepared the antibody specific for P-seq. By immunocytochemical methods with this and the other monoclonal antibody to cardiac C-protein (C-315), we found that two isoforms were coexpressed and colocalized in both atrial and ventricular muscles through development, but type I was scarcely expressed in embryonic skeletal muscle. These observations were confirmed by a RT-PCR method.
3) Competition of C-protein (MyBP-C) with H-protein (MyBP-H) for binding to myosin in cardiac muscle cells
C-Protein and H-protein are myosin-binding proteins which localize in A-band regions of sarcomeres. To clarify functional relationship of these two myosin binding proteins, we introduced cDNAs encoding these proteins into primary cultures of chicken myocytes and examined their assembly into myofibrils. The results obtained indicate that overexpression of MyBP-C or MyBP-H in cultured cardiac myocytes can reduce the content of MyBP-H or MyBP-C, respectively. These data indicate that MyBP-H can compete with MyBP-C for binding to myosin in the cells. Less

  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Nagaoka, Rie: "Quantitative and analysis of low MW-actin-binding proteins, cofilin, ADF and profilin, expressed in developing and degenerating chicken skeleta muscles." J.Muscle Res Cell Moeil.17. 463-473 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hayakawa, Kumihide: "Differential assembly of cytoskeletal and sarcomeric actins in developing skeletal muscle cells in vitor" Zool, Sci. 13. 509-517 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Endo, Takeshi: "Distinct troponin T genes are expressed in embyonic/larval tail striated muscle and adult body wall smooth muscle in ascidian" J.Biol. Chem.271. 27855-27862 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Obinata, Takashi: "Low molecular-waight G-actin binding proteins mvolved in the regulation of actin assembly during myofibrillogenesis." Cell. Str. Func.22(1). 181-189 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Begum, S.: "Dfferentiation of muscle-specific proteins in chicken somites as studied by immunofluorescence microscopy." Cell. Tiss. Res.293. 305-311 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soeno, Yoshinori: "Effects of exogenous β-actinin (CapZ) on actin filamentous structures in cultured muscle cells." Zool. Sci.15. 217-222 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Seno, Yoshinori: "Generation of functional β-actinin (CapZ) in an E.coli expression system" J. Muscle Res. Cell Motil.19(6). 639-646 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Vohra, M.S.: "Subcellular localization of dystrophin and vinculin in cardiac muscle fibers of the conduction system of the chicken ventricle." Cell Tiss. Res.294. 137-143 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kurasawa, Mariko: "Differential expression of C-protein isoforms in developing and degeneraling mousse striated muscles." Muscle & Nerve. 22. 196-207 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soeno, Yoshinori: "BDM (2,3-Butanedione monoxime), an inhibitor for myosin-actin interaction, suppresses myofibrillogenesis in skeletal muscle cells in culture." Cell Tiss. Res.295. 307-316 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kusano, Ken-ichi: "Datection of a suquensce involved in actin-binding and phospho-inositide-binding in the N-terminal side of cofilin." Mol. Cell. Biochem.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soeno, Yoshinori: "Organization of connectin/titin filaments in sarcomeres of differentiating chicken skeletal muscle cells." Mol. Cell. Biochem.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kusano, Ken-ichi: "Detection of a sequence involved in actin-binding and phospho-inositide-binding in the N-terminal side of cofilin." Mol. Cell. Biochem.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Soeno, Yoshinori: "Organization of connectin/titin filaments in sarcomeres of defferentiating chicken skeletal muscle cells." Mol. Cell. Biochem.in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nagaoka, R., Minami, N., Hayakawa, K., Abe, H.and Obinata, T.: "Quantitative analysis of low MW-actin-binding proteins, cofilin, ADF and profilin, expressed in developing and degenerating chicken skeletal muscles." J.Muscle Res Cell Motil.17. 463-473 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hayakawa, K., Ono, S., Nagaoka, R., Saitoh, O.and Obinata, T.: "Differential assembly of cytoskeletal and sarcomeric actins in developing skeletal muscle cells in vitro." Zool.Sci.13. 509-517 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Endo, T., Matsumoto, K., Hama, T., Ohtsuka, Y., Katsura, G.and Obinata, T.: "Distinct troponin T genes are expressed in embryonic/larvel tail striated muscle and adult body wall smooth muscle in ascidian." J.Biol.Chem.271. 27855-27862 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Obinata, T., Nagaoka-Yasuda, R., Ono, S., Kusano, K., Mohri, K., Ohtaka, Y., Yamashiro, S., Okada, K., and Abe, H.: "Low molecular-weight G-actin binding proteins involved in the regulation of actin assembly during myofibrillogenesis." Cell Str.Func.22 (1). 181-189 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soeno, Y., Hayakawa, K., and Obinata, T.: "Effects of exogenous b-actinin (CapZ) on actin filamentous structures in cultured muscle cells." Zool.Sci.15. 217-222 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soeno, Y., Abe, H., Kimura, S., Maruyama, K.and Obinta, T.: "Generation of functional b-actinin (CapZ) in an E.coli expression system." J.Muscle Res Cell Motil.19. 639-646 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Begum, S., Komiyama, M., Toyota, N., Obinata, T., Maruyama, K.and Shimada, Y.: "Differentiation of muscle-specific proteins in chicken somites as studied by immuno-fluorescence microscopy." Cell Tiss.Res.293. 305-311 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Vohra, M.S., Komiyama, M., Hayakawa, K., Obinata, T.and Shimada, Y.: "Subcellular localization of dystrophin and vinculin in cardiac muscle fibers of the conduction system of the chicken ventricle." Cell Tiss. Res.294. 137-143 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soeno, Y., Shimada, Y.and Obinata, T.: "BDM (2.3-Butanedione monoxime) an inhibitor for myosin-actin interaction, suppresses myofibrillogenesis in skeletal muscle cells in culture." Cell Tiss.Res.295. 307-316 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kurasawa, M., Sato, N., Matsuda, A., Koshida, S., Totsuda, T.and Obinata, T.: "Differentiation expression of C-protein isoforms in developing and degenerating mouse striated muscles." Muscle & Nerve. 22. 196-207 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Soeno, Y., Yajima, H., Kawamura, Y., Kimura, S., Maruyama, K.and Obinata, T.: "Organization of connectin/titin filaments in sarcomeres of differentiating chicken skeletal muscle cells." Mol.Cell Biochem.(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kusano, K., Abe, H.and Obinata, T.: "Detection of a sequence involved in actin-binding and phosphoinositide-binding in the N-terminal side of cofilin." MOl.Cell.Biochem.(in press). (1999)

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

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Published: 1999-12-08  

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