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1995 Fiscal Year Final Research Report Summary

Calcium-dependent protein kinase in osmoregulation of the halotolerant green alga Dunaliella tertiolecta

Research Project

Project/Area Number 06454013
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 植物生理
Research InstitutionNagoya University

Principal Investigator

TORIYAMA Shoshi  Nagoya University, Bioscience Center, Pofessor, 生物分子応答研究センター, 教授 (40013338)

Project Period (FY) 1994 – 1995
KeywordsCalcium-dependent protein kinase / Nucleotide sequence of cDNA / Osmoregulation / Dunaliella / Calcium ion / Osmotic-shock responsive protein kinase / Aequorin-transformed tobacco cell
Research Abstract

Calcium-dependent protein kinase (CDPK)was purified 20,000-fold from soluble franction of Dunaliella, and characterized. Using an anti-Dunaliella CDPK antibody, cDANs encoding parts of CDPK were cloned from a Dunaliella cDNA library. A full length cDNA was isolated by using one of partial cDNAs as a probe, and is nucleotide sequence was determined. Deduced amino acid squence from the nucleotide sequence indicated that Dunaliella CDPK in a typical CDPK which has kinase domain at N-terminus, calmolulin-like domain at C-terminus and pseudosubstrate domain between them.
When subjected to hypo-/hypersomotic shock, 40-kDa protein kinases named HAP/LAP kinases, respectively, which have different substrate specificities and time courses of activation were activated. The activation accompanied with protein phosphorylation. Since an involvement of increase in cytosolic Ca^<2+> was anticipated for the activation, tobacco suspension culture cells transformed aequorin was subjected osmotic shock. Upon hypoosmotic shock, a transient increase in Ca^<2+> with a peak at 70 sec after the shock and subsequent activation of 50-, 75- and 80-kDa protein kinases was observed. The activation accompanied with protein phosphorylation suggesting the involvement of CDPK.
A CDPK cross-reacting with anti-Dunaliella CDPK was detected in a brackish water Characeae Lamprothamnium. When this alga injected the antibody into the cytoplasm was subjected to hypoosmotic shock, turgor regulation was suppressed even though an increase in cytosolic Ca^<2+> occured after the shock. This result suggests the involvement of CDPK in turgor regulation of this alga.
The results obtained in this study suggest that CDPK is tightly involved in osmotic signal transduction and osmoregulation in algae and higher plants.

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] 湯浅高志、武藤尚志: "カルシウム依存性プロテインキナーゼ(CDPK):緑色植物と原生動物に特異的に存在、緑藻ドナリエラの浸透圧制御に関与?。" 化学と生物. 32. 489-491 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Yuasa,K.Takahashi and S.Muto: "Purification and characterization of a Ca^<2+>-dependent protein kinase from the halotolerant green alga Dunaliella tertiolecta." Plant Cell Physiol.36. 699-708 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Takahashi,T.Yuasa and S.Muto: "Purification and characterization of an algal(Dunaliella tertiolecta)protein cross-reacting with an anti-G_<α-common> antibody." Physiol.Plant.94. 486-490 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Yuasa and S.Muto: "Activation of 40-kDa protein kinases in response to hypo-and hyperosmotic shock in the halotolerant green alga Dunaliella tertiolecta." Plant Cell Physiol.37. 35-42 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 武藤尚志、湯浅高志、高橋宏二: "浸透圧調節とプロテインキナーゼ" 現代化学. (増刊),(印刷中). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 武藤尚志: "セカンドメッセンジャー" 現代化学. (増刊),(印刷中). (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Yuasa, K.Takahashi and S.Muto: "Purification and characterization of a Ca^<2+>-dependent protein kinase from the halotolerant green alga Dunaliella tertiolecta." Plant Cell Physiol. 36. 699-708 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Takahashi, T.Yuasa and S.Muto: "Purification and characterization of an algal (Dunaliella tertiolecta) protein cross-reacting with an anti-G_<alpha-common> antibody." Physiol.Plant.94. 486-490 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Yuasa and S.Muto: "Activation of 40-kDa protein kinases in response to hypo- and hyperosmotic shock in the halotolerant green alga Dunaliella tertiolecta." Plant Cell Physiol.37. 35-42 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S.Muto, T.Yuasa and K.Takahashi: "Protein kinases in osmolegulation. (in Japanese)" Gendai Kagaku (Modern Chemistry) Special issue. (in press). (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Muto: "Second messengers. (in Japanese)" Gendai Kagaku (Modern Chemistry) Special issue. (in press). (1996)

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

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Published: 1997-03-04  

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