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Cloning of a gene responsible for the cellular sensitivity to environmental stresses

Research Project

Project/Area Number 07680574
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 環境影響評価(含放射線生物学)
Research InstitutionUniversity of Tokyo

Principal Investigator

SAKAI Kazuo  University of Tokyo, Graduate School of Medicine, Lecturer, 大学院・医学系研究科, 講師 (40153837)

Co-Investigator(Kenkyū-buntansha) HIRANO Kazuya  Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (80251221)
ITOH Masamitsu  Graduate School of Medicine, Research Associate, 大学院・医学系研究科, 助手 (80176362)
SUZUKI Norio  Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (10010050)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1996: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
KeywordsHeat sensitivity / Heat resistance / Cell fusion / Gene transfection / Gene cloning / Dye exclusion test / DNA ladder
Research Abstract

To isolate and characterize a gene responsible for heat resistance/sensitivity in mammalian cells, hubrid clones were prepared from human glioma A7 cells, which are heat resistant, and Chinese hamster V79 cells, which are relatively heat sensitive. Subcloning of the most resistant hybrids, VA41, was carried out ; a subclone which was as resistant as 47, VA41-4, and a subclone which was as sensitive as V79, VA41-8, were obtained. The number of chromosomes in VA41-4 was larger than that in VA41-8. Human repetitive sequence was found, by Southern analysis, in VA41-4 as well as VA41-8, suggesting that a human gene responsible for heat resistance in VA41 and that VA41-8 became sensitive because it lost the gene during subculture. In order to identify the gene, a DNA transfection experiment, VA41-4 DNA into V79, was carried out. One of the transfectants, I41, was as resistant as A7. A genomic DNA library was prepared in EMBL4 phage vectors. Clones containing human repetitive sequences were found at a frequency of 5 x 10-4 in the library.
Also, cellular heat responses in A7, V79, and the hybrid clones were characterized in order to get clues for possible roles of the heat resistant gene.
The hypersensitivity to heat treatment seemed associated with DNA ladder formation and quick cell death. In heat sensitive V79 and VA41-8 DNA ladder found immediately after heat treatment, and dead cells appeared quickly after heat. On the other hand, heat resistant A7 and VA41-4 cell death process was slower and DNA ladder was not observed. These results suggest that the gene would make cells heat resistant by suppressing DNA ladder formation and cell death process.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Morimatsu, A.: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia." Journal of Radiation Research. 37(1). 1-11 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matusmoto, Y.: "DNA-dependent protein kinase activity of ataxia telangiectasia-and X-linked agammaglobulinemia-derived cell." Journal of Radiation Research. 37(4). 344-344 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kang, Y.: "Flowcytometric analysis of MUCI mucin production on the cell surface in human carcinoma cell lines." Journal of Radiation Research. 37(4). 361-361 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K.: "Heat sensitivity of hybrid clones and its modification." Journal of Radiation Research. 37(4). 391-391 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kang, Y.: "Characterization of the expression of MUC1 mucin in human colon carcinoma HT29 cells after exposure to X-ray." Journal of Radiation Research. 38(4). 330-330 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Enomoto, A.: "Characterization of radiation-hybrid clones isolated from human MOLT-4 and mouse FM3A cells." Journal of Radiation Research. 38(4). 339-339 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K.: "Effect of Zn^<2+> on the radio-and heat sensitivity of cultured mammalian cells." Journal of Radiation Research. 38(4). 353-353 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto, Y.: "A possible mechanism for hyperthermic radiosensitization mediated through hyperthermic lability of Ku subunits in DNA-dependent protein kinase." Biochemical and Biophysical Research Communications. 234. 568-572 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K.: "Effects of an inhibitor of protein kinases on the response to heat treatment in cultured mammalian cells." International Journal of Hyperthermia.13(5). 535-545 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K., Yokogawa, H., and Suzuki, N.: "Cellular heat sensitivity and the DNA ladder after heat treatment" J.Radiat.Res.36. 384-384 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Morimatsu, A., Suzuki, N., Hirano, K., Matsumoto, Y., and Sakai, K.: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia." J.Radiat.Res.37. 1-11 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto, Y,Naritaka, S., Sakai, K., and Suzuki, N.: "DNA-dependent protein kinase activity of ataxia telangiectasia-and X-linked agammaglobulinemia-derived cells." J.Radiat.Res.37. 344-344 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kang, Y., Hirano, K., Sakai, K., Irimura, T., and Suzuki, N.: "Flowcytometric analysis of MUC1 mucin production on the cell surface in human carcinoma cell lines." J.Radiat.Res.37. 361-361 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K., and Suzuki, N.: "Heat sensitivity of hybrid clones and its modification." J.Radiat.Res.37. 391-391 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kang, Y., Hirano, K., Sakai, Suzuki, N., and Irimura, T.: "Characterization of the expression of MUC1 mucin in human colon carcinoma HT29 cells after exposure to X-ray." J.Radiat.Res.38. 330-330 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Enomoto, A., Sakai, K., Hirano, K., Suzuki, N., and Koyama, H.: "Characterization of radiation-hybrid clones isolated from human MOLT-4 and mouse FM3A cells." J.Radiat.Res.38. 339-339 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K., and Suzuki, N.: "Effect of Zn2+ on the radio-and heat sensitivity of cultured mammalian cells." J.Radiat.Res.38. 353-353 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai, K,Suzuki N,Itoh H and Kubodera A: "Effect of an inhibitor of protein kinases on the response to heat treatment in cultured mammalian cells." Int.J.Hyperthermia. 13(5). 535-545 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumoto, Y., Suzuki, N., Sakai, K., Morimatsu, A., Hirano, K., and Murofushi, H.: "A possible mechanism for hyperthermic radiosensitization mediated through hyperthermic lability of Kusubunits in DNA-dependent protein kinase." Biochem.Biophys.Res.Commun.234. 568-572 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakai,K.: "Effects of an inhibitor of protein kinases on the response to heat treatment in cultured mammalian cells." International Journal of Hyperthermia.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Matusmoto,Y.: "Apossible mechanism for hyperthermic radiosensitization mediated through hyperthermic lability of Ku subumits in DNA-dependent protein kinase." Biochemical and Biophysical Research Communications. (in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Morimatsu,A.: "Identification and characterization of a protein appeared after X-irradiation in human T cell leukemia." Journal of Radiation Research. 37. 1-11 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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