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1987 Fiscal Year Final Research Report Summary

Base changes in the replication of DNA induced by nucleotide analogs and the use for in vitro mutagenesis

Research Project

Project/Area Number 61480430
Research Category

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

Allocation TypeSingle-year Grants
Research Field Biological pharmacy
Research InstitutionOkayama University

Principal Investigator

HAYATSU Hikoya  Faculty of Pharmaceutical Sciences, Okayama University, 薬学部, 教授 (10012593)

Co-Investigator(Kenkyū-buntansha) NEGISHI Kazuo  Faculty of Pharmaceutical Sciences, Okayama University, 薬学部, 助手 (70116490)
WATAYA Yusuke  Faculty of Pharmaceutical Sciences, Okayama University, 薬学部, 助教授 (90127598)
Project Period (FY) 1986 – 1987
KeywordsN^4-Aminocytidine / transition mutation / replication error / M13mp2,FM3A cell / V79 cell / トランジション変異 / 複製
Research Abstract

The moecular mechanism of N^4-aminocytidine-mediated mutagenesis was investigated. A summary of findings achieved is as follows. (1) A study of in vitro DNA synthesis using E. coil DNA polymerase I large fragment as the enzyme showed that N^4-aminodeoxycytidine 5'-triphosphate (dC^<am>TP) can serve as a substrate for the enzyme and can be incorporated int onewly formed DNA. The efficiency of incorporation was about 50% of the natural substrate dCTP, and this nucleotide analog can be used as a substitute for dCTP and dTTP. (2) Use of dC^<am>TP in the M13mp2 DNA synthesizing system, followed by transfection of the resulting DNA to E. coli allowed production of mutant phages. More than 100 mutant clones were spearated, the DNA was isolated and their nucleotide sequences determined. The results indicated that all possible transitions, A to G, G to A, C to T and T to C, and only these transitions, have occurred. This result is consistent with a mechanism in which tautomeric shift in N^4-aminocytosine between the amino and the imino forms is the cause of the mutation. (3) Mutations were induced in Drosophila when the larve were fed with N^4-aminocytidine. (4) An improved method to synthesize N^4-aminocytidine was established.

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] M. Takahashi;K. Negishi and H. Hayatho: Biochim. Biophys. Res. Commun.143. 104-109 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A. Nomura;K. Negishi;H. Hayatsu and Y. Kuroda: Mutation Res.177. 283-287 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K. Negishi;M. Kawakami;K. Kayasuga;J. Odo and H. Hayatsi: Chem. Pharm. Bull.35. 3884-2887 (1987)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M. Tadahashi;M. Nishizawa;K. Negishi;F. Hanaoka;M. Yamada and H. Hayatsu: Mol. Cell. Biol.8. 347-352 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Negishi;K. Negisho;H. Ryo;S. Kondo and H. Hayatsu: Mutagenesis. 3. 11-13 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K. Negisho;K. Matsumoto;T. Bassho;F. Tada and H. Hayatsu: Nucleic Acids Symposium Series No. 19. 33-26 (1988)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M. Takahashi, K. negishi and H. hayatsu: "Proofreading of a mutagenic nucleotide, N^4-aminodeoxycytidylic acid, by Escherichia coli DNA polymerase I." Biochem. Biophys. Res. Commun.143. 104-109 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Nomura, K. Negishi, H. Hayatsu and Y. Kuroda: "Mutagenicity of N^4-aminocytidine and its derivatives in Chinese hamster lund V79 cells. Incorporation of N^4-cminocytosine into cellular DNA" Mutation Res.177. 283-287 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Negishi, M. kawakami, K. kayasuga, J. Odo and H. Hayatsu: "An improved synthesis of N^4-aminocytidine" Chem. Pharm. Bull.35. 3884-3887 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M. Takahashi, M. Nishizawa, K. Negishi, F. Hanaoka, M. Yamada and H. hayatsu: "Induction of mutation in mouse FM3A cells by N^4-aminocytidine mediated replicational errors." Mol. Cell. Biol.8. 347-352 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Negishi, K. Negishi, H. Ryo, S. Kondo and H. Hayatsu: "The genotoxicity of N^4-aminocytidine in the Drosophila wing spot test." Mutagenesis. 3. 11-13 (1988)

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

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Published: 1989-03-30  

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