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Development of p53-associated molecular devices for diagnosis and treatment of gastrointestinal cancer

Research Project

Project/Area Number 14370173
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Gastroenterology
Research InstitutionTohoku University

Principal Investigator

ISHIOKA Chikashi  Tohoku University, Institute of Development, Aging and Cancer, Professor, 加齢医学研究所, 教授 (60241577)

Co-Investigator(Kenkyū-buntansha) SAKAYORI Masato  Tohoku University, Hospital, Research Associate, 病院・助手 (20344666)
KATO Shunsuke  Tohoku University, Hospital, Research Associate, 病院・助手 (60333887)
SHIBATA Hiroyuki  Tohoku University, Hospital, Lecturer, 病院・講師 (50260071)
Project Period (FY) 2002 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,000,000 (Direct Cost: ¥14,000,000)
Fiscal Year 2003: ¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 2002: ¥8,600,000 (Direct Cost: ¥8,600,000)
Keywordsp53 / gastrointestinal cancer / super p53 / second-site suppressor
Research Abstract

The objectives of this study were (i) development of p53-associated molecular devices for diagnosis of gastrointestinal cancer, including novel method of detecting p53 gene mutations and mutant p53 proteins, and (ii) development of p53-associated molecular devices for treatment of gastrointestinal cancer, including improved p53 cDNA for p53-mediated gene therapy and small molecule compounds for restoration of inactivated mutant p53 function. We developed several important p53-related molecular devices. First, by using a comprehensive site-directed mutagenesis technique and a yeast-based functional assay to construct, express, and evalute 2,314 p53 mutants representing all possible amino acid substitutions caused by a point mutation throughout the protein (5.9 substitutions per residue), and cerrelated p53 function with structure-and tumor-derived mutations. Second, to identify key temperature-sensitive (ts) structural elements controlling the protein function, we screened ts p53 mutant … More s from a comprehensive mutation library consisting of 2,314 p53 missense mutations for their sequence-specific transactivity through p53-binding sequences in Saccharomyces cerevisiae. We isolated 142 ts p53 mutants, including 131 unreported ts mutants and showed that the intramolecular beta-sheet in the core DNA-binding domain of p53 was a key structural element controlling the rotein function and provided a clue for finding a molecular mechanism that enables the rescue of the mutant p53 function. Third, by using the p53 mutation library, we identified second-site suppressor (sss) mutations for common p53 mutations. The identified sss mutations restored two of the common mutations by both intramolecular and intermolecular manner. The results suggested that inactivated function of specific p53 mutants might be recovered by extramolecules. Fourth, as a result of intensive screening of the p53 mutation library, we identified at least several mutant p53 that had the stronger activity to induce apoptosis than wild-type p53. These mutant p53 might be useful cDNA resources for cancer gene therapy. Finally, we are screening in silico of small molecular compounds that might restore the inactivated function of mutant p53 through a molecular docking simulation method, and found several candidate molecules. Less

Report

(3 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Kate S, et al.: "Understanding the function-structure and function-mutation relationships of p53 tumor suppresser protein by high resolution missense mutation analysis."Proc.Natl.Acad.Sci., USA. 100. 8424-8429 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shiraishi K, et al.: "Isolation of temperature-sensitive p53 mutations from a comprehensive missense mutation library."J.Biol.Chem.. 279. 348-355 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Otsuka K, et al.: "Analysis of the human APC mutation apectrum in a Saccharomyces cerevisiae strain with a mismatch repair defect"International Journal of Cancer. 103. 624-630 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakayori M, et al.: "Evaluation of the diagnostic accuracy of the stopn codon (SC) assay for identifying protein-truncating mutations in the BRCA1 and BRCA2 genes in familial breast cancer"Journal of Human Genetics. 48. 130-137 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Shiraishi, K., Kato, S., Han, S., Liu, W., Otsuka, K., Sakayori, M., Ishida, T., Takeda, M., Kanamaru, K., Obuchi, N., Ishioka, C.: "Isolation of Temperature-sensitive p53 Mutations from a Comprehensive Missense Mutation Library."J Biol Chem. 279. 348-355 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kato, S., Han, S., Liu, W., Otsuka, K., Shibata, H., Kanamaru, R., Ishioka, C.: "Understanding the function-structure and function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis."Proc Natl Acad Sci USA. 100. 8424-8429 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Sakayori, M., Kawahara, M., Shiraishi, K., Nomizu, T., Shimada, A., Kudo, T., Abe, R., Ohuchi, N., Takenoshita, S., Kanamaru, R., Ishioka, C.: "Evaluation of the diagnostic accuracy of the stop codon (SC) assay for identifying protein-truncating mutations in the BRCA1 and BRCA2 genes in familial breast cancer."J Hum Genet. 48. 130-137 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Otauka, K., Suzuki, T., Shibata, H., Kato, S., Sakayori, M., Shimodaira, H., Kanamaru, R., Ishioka, C.: "Analysis of the human APC mutation spectrum in a saccharomyces cerevisiae strain with a mismatch repair defect."Int J Cancer. 103. 624-630 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] Kato S, et al.: "understanding the function-structure and function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis"Proc.Natl.Acad.Sci., USA. 100. 8424-8429 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shiraishi K, et al.: "Isolation of temperature-sensitive p53 mutations from a comprehensive missense mutation library"J.Biol.Chem.. 279. 348-355 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Tsuchida R, et al.: "Detection of ATM gene mutation in human glioma cell line M059J by a rapid frameshift/stop codon assay in yeast"Radiation Research. 158. 195-201 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Otsuka K, et al.: "Analysis of the human APC mutation spectrum in a accharomyces cerevisiae strain with a mismatch repair defect"International Journal of Cancer. 103. 624-630 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sakayori M, et al.: "Evaluation of the diagnostic accuracy of the stopn codon (SC) assay for identifying protein-truncating mutations in the BRCA1 and BRCA2 genes in familial breast cancer"Journal of Human Genetics. 48. 130-137 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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