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2003 Fiscal Year Final Research Report Summary

Development of anti-virus agents on the basis of their ability to induce replicational errors

Research Project

Project/Area Number 14572093
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 医薬分子機能学
Research InstitutionOKAYAMA UNIVERSITY

Principal Investigator

NEGISHI Kazuo  Okayama University, Advanced Science Res. Ctr., Associate Professor, 自然生命科学研究支援センター, 助教授 (70116490)

Co-Investigator(Kenkyū-buntansha) OTSUKA Chie  Okayama University, Advanced Science Res. Ctr., Research Associate, 自然生命科学研究支援センター, 助手 (60253001)
TOYODA Tetsuya  Kurume University, School of Medicine, Professor, 医学部, 教授 (00197972)
Project Period (FY) 2002 – 2003
Keywordsnucleotide analogs / error catastrophe / replicational error / ambiguous base-pairing / RNA virus / retrovirus / RNA polymerase / HPLC
Research Abstract

1. Synthesis of nucleotide analogs.
We have established the methods for synthesis of N^4-hydroxycytidine triphosphate (N^4-OhCTP),N^4-methoxycytidine (N^4-MoCTP), and N^4-methycytidine (N^4-McCTP), and obtained pure materials. With these pure, samples, their incorporation into RNA catalyzed by T3 RNA polymerase was studied. RNA synthesis took place if N^4-OhCTP, or N^4-MoCTP was present in place of UTP, but not when they were added in place of CTP. This results shows that they behave only as UTP analogs in spite of their ambiguity in bass-pairing. In contrast, N^4-MeCTP behaved as CTP analog only. Incorporation properties of isoGTP, one of typical oxidation products from nucleotide, was studied using a commercial sample. It shows no incorporation by the RNA polymerase.
2.Properties of the analogs in templates in the reversetranscription.
Above finding enabled us to make RNA whose U or C was totally replaced with the analogs. Thus we have prepared RNA with analogs, and they were used as a template for reverse transcription with AMV reversetranscriptase. Resulting cDNA was amplified and analyzed with a DNA sequencer. The analysis indicated that N^4-hydroxycytidine was read as C and T almost equally. It suggests that this analog may cause mutation in a reversetranscription process.
3. Anti-HIV virus activities of the analogs.
Some analogs ware found to be very hopeful. Further study is continuing.
4. Effects on RNA viruses and their polymerase.
Action of analog triphosphates to Polio RNA polymerase was studied.
5. Use of chitosan-triphosphate complexes.
We have found that chitosan-triphosphate complex made by a reported method was not stable enough. Improvement of the protocol is currently underway.

  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] Chie Otsuka, Kazuo Negishi他: "Difference between deoxyribose- and tetrahydrofuran-type abasic sites in the in vivo mutagenic responses in yeast"Nucleic Acids Research. 30. 5129-5135 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chie Otsuka, Kazuo Negishi他: "Use of yeast transformation by oligonucleotides to study DNA lesion bypass in vivo"Mutation Research. 502. 53-60 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hikoya Hayatsu, Kazuo Negishi, Masahiko Shiraishi: "Acceleration of Bisulfite-Mediated Deamination of Cytosine in DNA"Proceedings of the Japan Academy. (In press). (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kei Moriyama, Toshiyuki Okada, Kazuo Negishi: "Mutagenic Nucleoside and Nucleotide Analogues, and a Novel Approach to anti-HIV Drug Therapy In "Recent Resrch Developments in organic and Bioorganic Chemistry""Research Signpost, Kerala, India. 10 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chie Otsuka, Kazoo Negishi, others: "Difference between deoxyribose-and tetrahydrofuran-type abasic sites in the in vivo mutagenic responses in yeast"Nucleic Acids Research. 30. 5129-5135 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chie Otsuka, Kazuo Negishi, others: "Use of yeast transformation by oligonucleotides to study DNA lesion bypass in vivo"Mutation Res.. 502. 53-60 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hikoya Hayatsu, Kazuo Negishi, Masahiko Shiraishi: "Acceleration of Bisulfite Mediated Deamination of Cytosine in DNA"Proc.Japan Acad.2004. (in press). (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kei Moriyama, Toshiyuki Okada, Kazuo Negishi: "Mutagenic Nucleoside and Nucleotide Analogues, and a Novel Approach to anti-HIV Drug Therapy"Recent Research Developments is Organic and Bioorganic Chemistry, Research Signpost, Kerala, lndia, 2002. 152-161 (2002)

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

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Published: 2005-04-19  

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