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Identification of a growth-suppressing gene in normal human fibroblasts and mouse brain

Research Project

Project/Area Number 03454556
Research Category

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

Allocation TypeSingle-year Grants
Research Field 分子遺伝学・分子生理学
Research InstitutionInstitute of Molecular and Cellular Biosciences

Principal Investigator

AYUSAWA Dai  Institute of Molecular and Cellular Biosciences, Associate Professor, 分子細胞生物学研究所, 助教授 (00142109)

Co-Investigator(Kenkyū-buntansha) OISHI Michio  Institute of Molecular and Cellular Bioscience, Professor, 分子細胞生物学研究所, 教授 (00126004)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1991: ¥2,900,000 (Direct Cost: ¥2,900,000)
KeywordsSenescence / Immortalization / Gene / Chromosome 7 / Signal transduction / Gene mapping / L1反復配列 / 相補性群 / 不死化細胞株 / 微小核細胞融合法 / 放射線雑種細胞 / 分裂停止遺伝子 / 老化遺伝子 / 正常線維芽細胞 / 不死化 / RFLP解析 / 微少核細胞融合法 / 不死化遺伝子 / 染色体導入 / 染色体マッピング / 不死化細胞 / アンチセンスcDNAライブラリ-
Research Abstract

Normal somatic cells in mammals stop dividing in culture after a limited number of cell division. This phenomenon is called cellular senescence and governed by several genes. When these genes are inactivated in a cell, it acquires potential to divide indefinitely and immortalize. Genetic analysis revealed that there exist at least four genetic complementation groups (A-D) in immortal human cell lines. We have found that introduction of human chromosme 7 by microcell-mediated chromosome transfer specifically induces senescence in cell lines belonging to group D.Then we made a panel of radiation hybrids under the backgroud of mouse A9 cells. The hybrids were characterized by FISH analysis and PCR with STS markers to determine how fragments of human chromosme 7 are integrated on mouse A9 genome. Of these hybrids, we selected a clone that contains only 2-3 Mb human DNA and can induce senescence in group D cell lines. We rescued human DNA from this clone and used it as a probe to map the putative senescence gene on human chromosme 7. The mapping data are useful for us to identify the gene.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] D.Ayusawa et al.: "Mouse FM3A cell-cycle mutant with thermolabile ubiquitin-activating enzyme E1,as arrested in the S-phase and corrected by a cloned human E1 cDNA." Cell Struc.Func.17. 113-122 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Kaneda et al.: "Promoter-specific regulatory sequences in the human thymidylate synthase gene clustered at the 5,-end of the first intron." Somat.Cell Mol.Genet.18. 409-415 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] N.Imai et al.: "Cloning and sequence of a functionally active cDNA for mouse ubiquitin activating enzyme E1." Gene. 118. 282-297 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Takahash et al.: "A restriction fragment length polymorphism (RELP) locus of rat (rattus norvegicus) linkedto the Cs-1 locus of rat group XIII." Biochemical Genet.30. 537-544 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Ogata et al.: "Introduction of chromosome 7 suppresses indefinite growth in physicochemically immortalized human fibroblast line KMST-6 and SUSM-1." Mol.Cel.Biol.13. 6036-6043 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Hara et al.: "The UDP-galactose translocator gene is mapped to the Wiskott-Aldrich syndrome locuc of the X chromosome." Semat.Cell Mol.Genet.19. 571-575 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Mishina, D.Ayusawa, T.Seno and H.Koyama.: "Thymidylate stress induces homologous recombination activity in mammalian cells." Mutation Res.246. 215-220 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] D.Ayusawa, S.Kaneda, Y.Itoh, H.Yasuda, Y.Murakami, K.Sugawara, F.Hanaoka and T.Seno.: "Mouse FM3A cell-cycle mutant with thermolabile ubiquitin-activating enzyme E1, as arrested in the S-phase and corrected by a cloned human E1 cDNA." Cell Struc.Func.17. 113-122 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Kaneda, N.Horie, K.Takaishi, A.Takayanagi, T.Senoand D.Ayusawa.: "Promoter-specific regulatory sequences in the human thymidylate synthase gene clustered at the 5'-end of the first intron." Somat.Cell Mol.Genet.18. 409-415 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] N.Imai, S.Keneda, Y.Nagai, T.Seno, D.Ayusawa, F.Hanaoka and F.Yamao.: "Cloning and sequence of a functionally active cDNA for mouse ubiquitin activating enzyme E1." Gene. 118. 297-282 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] M.Takahashi, H.Yokota, D.Ayusawa, M.Oishi and T.Kunieda.: "A restriction fragment length polymorphism (RELP) locus of rat (rattus norvegicus) linkedto the Cs-1 locus of rat group XIII." Biochemical Genet.30. 537-544 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Ogata, D.Ayusawa, M.Namba, E.Takahashi, M.Oshimura and M.Oishi.: "Introduction of chromosome 7 suppresses indefinite growth in physicochemically immortalized human fibroblast line KMST-6 and SUSM-1." Mol.Cel.Biol.13. 6036-6043 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Hara, T.Takahashi, M.Yamauchi, M.Hoshino, K.Aoki, T.Hori, D.Ayusawa and M.Kawakita.: "The UDP-galactose translocator gene is mapped to the Wiskott-Aldrich syndrome locuc of the X chromosome." Semat.Cell Mol.Genet.19. 571-575 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.F.Sasaki, D.Ayusawa and M.Oishi.: "Construction of a normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system." Nucleic.Acid.Res.22. 987-992 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.F.Sasaki, T.Imasaki, H.Kobayashi, S.Tsuji, D.Ayusawa and M.Oishi.: "Consruction of an equalized cDNA library from human brain by seemi-solid self-hybridization system." DNA Research. 1. 91-96 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] J.Fujiwara, T.Kimura, D.Ayusawa and M.Oishi.: "A novel regulatory sequences affecting the contitutive expression of the tissue plasminogen gene in human Bows cell." J.Biol.Chem.269. 18558-18562 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ogata et al.: "Chromosome 7 induces cellular senescence of non-tumorigenic immortalized human fibroblast cell lines" Mol.Cell Biol.13. 6036-6043 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Sasaki et al.: "Construction of a normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system" Nucleic Acids Res.(in press).

    • Related Report
      1993 Annual Research Report
  • [Publications] Takahashi,M.: "A restriction fragment length polymorphism (RFLP) locus of rat (rattus norvegicus) linked to the Cs-l locus of rat linkage group XIII." Biochem.Genet.30. 537-543 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Koyama,H.: "Gene correction by homologous recombination at the deleted aprt locus in CHO cells." Molecular Approaches to the Study and Treatment of Human Deseases.347-355 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takayanagi,A.: "Intron 1 an the 5'-flanking region of the human thymidylate synthase gene as a regulatory determinant of growth-dependent expression." Nucleic Acid Res.20. 4021-4025 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ayusawa,D.: "Complementation by a cloned human ubiquitin-activating enzyme El of the S-phase-arrested mouse FM3A cell mutant with thermolabile El" Cell Struc.and Func.17. 113-122 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Imai,N.: "Cloning and sequence of a functionally active cDNA encoding the mouse ubiquitin-activating enzyme El" Gene. 18. 279-282 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kaneda,S.: "Regulatory sequences clusterd at the 5'end of the firts intron of the human thymidylate synthase gene function in cooperation with the promoter region." Somat.Cell and Molec.Genet.18. 409-415 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takahashi,E.,Ayusawa,D.,Kaneda,S.,Itoh,Y.,Seno,T.& Hori,T.: "The human ubiquitinーactivating enzyme El gene(UBE1)mapped to band Xp.11.23ーp11.3 by fluorescence in situ hybridization." Cytogenet.Cell Genet.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kaneda,S.,Horie,N.,Takahashi,K.,Takayanagi,A.,Seno,T.& Ayusawa,D.: "Promoterーspecific regulatory sequence in the human thymidylate synthase gene clustered at the 5'ーend of the first intron." J.Biol.Chem.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ayusawa,D.,Kaneda,S.,Itoh,Y.,Yasuda,H.,Murakami,Y.,Sugawara,K.,Hanaoka,F.& Seno.: "Mouse FM3A cellーcycle mutant with thermolabile ubiquitinーactivating enzyme,as arrested in the 5ーphase and corrected by cloned human E1 cDNA" Cell Struc.Func.(1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Hara,T.,Takahashi,E.,Yamauchi,M.,Hoshino,M.,Aoki,K.,Hori,T.,Kawakita,M.& Ayusawa,D.: "The UDPーgalactose translocator gene is mapped to the WiskottーAldrich syndrome locus of the X chromosome."

    • Related Report
      1991 Annual Research Report
  • [Publications] Takayanagi,A.,Kaneda,S.,Ayusawa,D.& Seno,T.: "Intron 1 and 5'ーflanking region of the human thymidylate synthase gene as a determinant of cellーcycle dependent posttranscriptional regulation."

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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