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1993 Fiscal Year Final Research Report Summary

Study on the structure and function of a novel stressprotein

Research Project

Project/Area Number 03680186
Research Category

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

Allocation TypeSingle-year Grants
Research Field 放射線5生物学
Research InstitutionAichi Cancer Center Research institution

Principal Investigator

OHTSUKA Kenzo  Aichi Canser Center, Lab, Exptl. Radiol, . Senior Researcher, 放射線部, 主任研究員 (40150213)

Project Period (FY) 1991 – 1993
KeywordsHeat-shock protein / hsp40 / DnaJ homologue / Molecular chaperone
Research Abstract

We have previously reported that a novel 40-kDa protein is induced by heat shock and several envionmental stresses in mammalian and avian cells and that the N-terminal amino acid sequence of this 40-kDa protein has homology with the bacterialdna Jheat-shock protein. We nextisolated and characterized a cDNA clone encoding 40-kDa heat-shock protein hsp40 from cDNA expression library of human placentaby immunoscreening with antiidenticalto that of HeLacellhsp40 as determined by amino acid sequencer. Deduced amino acid sequence of the cDNA is homologous to bacterial DnaJheat-shock protein834% identical) and its homologues in yeast such as SCJ1, YDJ1(MAS5), SIS1, SEC63 AND Zuotin(34 - 40% identical), with SIS1 being the most homologous. Nucleotide sequence identity between hsp40 and HDJ1(another human DnaJ homologue9 is more than 98% including 5'and3' noncoding region, suggesting that these two proteins are the same geneproduct. Since there are at least three dnaJ homologues in human [hsp40 … More (HDJ1), HDJ2(HSDJ), HSJ1] and hsp90 family.
Our hsp40 has JandG/F domains but not C domain. Inyeast DnaJ homologues SIS1 has a domain structure and biochemical properties (MW, pl, heat-inducibility and intracellular localization) similar to human hsp40. Also, Therefor, it is suggested that human hsp40 is ahomologue of yeast SIS1 protein.
Indirect immunofluorescence revealed that hsp40 in Hela cells accumulates in the nuclei, especially in the nucleoli during heat shock and returns to the cytoplasm during recovery period. The kinetics of the accumulation in the nucleoli and subsequent return to the cytoplasm during recovery period. The kinetics of the accumulation hsp40 was colocalized with hsc70 (p73) in heat-shocked HeLa cells as demonstrated by double immunofluorescence staining. Hsp70 in the nucleoli was released by the addition of ATP but not by ADP, GTP, nonhydrolyzableATP, nor high saltbuffer. In contrast, hsp40 was not released from the nucleoli with any of these treatments or any combination of these treatments. Thus, hsp40 was not released from the nucleoli after hsp70 has been released in an ATP-dependent manner. These results suggest that hsp40 (DnaJaggregates in the nuclei and nucleoli of heat-shocked HeLa cells. Less

  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] 大塚健三: "Cloning of a cDNA for heat-shock protein hsp40,a human homologue of bacterial DnaJ." Biochem.Biophys.Res.Comm.197. 235-240 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大塚健三 他: "Effect of ATP on the release of hsp70 and hsp40 from the nucleus in heat-shocked HeLa cells." Exptl.Cell Res.209. 357-366 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 光富徹哉 他: "Transformation by viral oncogenes in creased sensitivity to heat in rat 3Y1 fibroblasts." Anticancer Res.13. 995-1000 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 服部浩明 他: "A stress-inducible 40-kDa protein(hsp40):Purification by modified two-dimensional gel electrophoresis and colocalijation with hsc70(p73)in heat-shocked HeLa cells." J.Cell Sci.104. 629-638 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大塚健三 他: "Cytoskeletal thermotolerance in NRK cells." Int.J.Hyperthermia. 9. 115-124 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 大塚健三: "DneK-DneJシャペロン系は普遍的な存在か?" 蛋白質・核酸・酵素. 38. 875-876 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Ohtuka: "Cloning of a cDNA for heat-shock protein hsp40, a human homologue of bacterial DnaJ." Biochem. Biophys. Res. Comm.197. 235-240 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Ohtsuka, K.R.Utsumi, T.Kaneda and H.Hattori: "effect of ATP on the release of hsp70 and hsp40 from the nucleus in heat-shocked HeLa cells." Exptl. Cell Res.209. 357-366 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T.Mitsudomi, Y.Kido, K.Ohtsuka and K.sugimachi: "Transformation by viral oncogenes increased sensitivity to heat in rat 3Y1 fibroblasts." Aiticancer Res.13. 995-1000 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H.Hattori, T.Kaneda, B.Lokeshwar, A.Laszio and K.Ohtsuka: "A stress-inducible 40-kDa protein(hsp40) : purification by modified two-dimensional gel electrophoresis and co-localization with hsc70(p73) in heat-shocked HeLa cells." J.Cell Sci. 104. 629-638 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Ohtsuka, Y.-C.Liu and T.Kaneda: "Cytoskelatal thermotolerance in NRK cells." Int.J.Hyperthermia. 9. 115-124 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Ohtsuka: "Is the DnaK-DnaJ chaperone system universal? (in Japanese)" Protein, Nucleic Acid and Enzyme. 38. 875-876 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K.Ohtsuka: "HSP40.(in Japanese)" Modern Med.Lab.21. 944-945 (1993)

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

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Published: 1995-03-27  

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