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Molecular mechanisms of the adaptive response to alkylating agents

Research Project

Project/Area Number 02044114
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionKYUSHU UNIVERSITY

Principal Investigator

SEKIGUCHI Mutsuo  Medical Institute of Bioregulation, Kyushu University, 生体防御医学研究所, 教授 (00037342)

Co-Investigator(Kenkyū-buntansha) NAKABEPPU Yusaku  Medical Institute of Bioregulation, Kyushu University, 生体防御医学研究所, 助手 (30180350)
SHIMIZU Kenji  Kyushu University School of Medicine, 医学部, 助教授 (10037286)
DAVID Ludlum  マサチューセッツ大学, 医学部, 教授
MICHAEL Volk  マサチューセッツ大学, 医学部, 準教授
VOLKERT Michael  University of Massachesetts, School of Medicine
LUDLUM David B.  University of Massachesetts, School of Medicine
作見 邦彦  九州大学, 生体防御医学研究所, 助手 (50211933)
VOLKERT Mich  マサチューセッツ大学, 医学部, 準教授
MATIJASEVIC ゼンカ  マサチューセッツ大学, 医学部, 研究員
早川 浩  九州大学, 医学部, 助手 (70150422)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥7,100,000 (Direct Cost: ¥7,100,000)
Fiscal Year 1992: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1991: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1990: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsDNA Repair Enzyme / DNA Damage / Transcriptional Control / Gene Expression / Mutation / Ada Regulon / Alkylating Agent / Adaptive Response / 突然変異の誘起 / 発がん / 抗がん剤 / メチルトランスフェラ-ゼ / DNA修復酸素
Research Abstract

Exposure of Escherichia coli cells to relatively low levels of alkylating agents leads to the formation of enzymes related to the repair of alkylation-associated DNA damage. Once these enzymes are produced, the cell becomes more resistant to both the killing and mutagenic effects of various alkylating agents. This process is termed the "adaptive response". In this process at least four genes, ada, alkA, alkB, and aidB, are known to be involved, among which the ada gene plays a central role. Ada protein, the product of the ada gene, acts as a transcriptional regulator for its own gene and for others belonging to this regulon. The Ada protein carries two distinct methyltransferase activities, one transfers a methyl group from one of two stereoisomers of methylphosphotriesters of alkylated DNA to a specific cysteine residue (Cys69) of the enzyme and the other transfers a methyl group from an O^6-methylguanine or O^4-methylhymine residue of the DNA to a cysteine residue (Cys321) located cl … More ose to the C-terminus of the protein.
Expression of the alkA gene in Escherichia coli is controlled by Ada protein, which binds to a specific region of the alkA promoter and enhances further binding of RNA polymerase holoenzyme to the complex. To determine the sequence recognized by the Ada protein, we introduced various base substitutions into the promoter region of alkA and examined their effects on expression of the gene, both in vivo and in vitro. Base changes within the sequence AAAGCAAA, located between positions -41 and -34 from the transcription initiation site, greatly decreased the frequencies of initiation of transcription. In footprinting experiments, the region containing this sequence was protected by the Ada protein and base change within this sequence let to failure of binding of Ada protein to the promoter. It is likely that the Ada protein recognizes the AAAGCAAA sequence in the alkA promoter and binds to the region containing the sequence, thereby allowing ready access of RNA polymerase to the promoter. There are considerable differences between the mechanisms of action of Ada protein on the promoters of alkA and ada, even though the expression of both genes is positively relulatoed by Ada protein. Less

Report

(3 results)
  • 1992 Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] 白石 明子,作見 邦彦,中津 可道,早川 浩,関口 睦夫: "cDNA and genomic sequences for mouse O^6-methylguanine-DNA methyltransferase" Carcinogenesis. 13. 289-296 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 闕 玲玲,中村 崇規,中津 可道,作見 邦彦,早川 浩,関口 睦夫: "Specific amino acid sequired for O^6-methylguanine-DNA methyltransferase activity:Analysis of three residues at or near the methyl acceptor site" Carcinogenesis. 13. 837-843 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Z.MATIJASEVIC,関口 睦夫,David B.Ludlum: "Release of N^2,3-ethenoguanine from chloroacetaldehydetreated DNA b Escherichia coli 3-methyladenine DNA glycosylase II" Proc.Natl.Acad.Sci.,U.S.A.89. 9331-9334 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 古市 正人,宇 成国,穴井 元昭,作見 邦彦,関口 睦夫: "Regulatory elements of expression of the alkA gene in response to alkylating agents" Molec.gen.Genet.236. 25-32 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 莫 錦尭,真木 寿治,関口 睦夫: "Hydrolytic elimination of a mutagenic nucleotide,8-oxodGTP,by human 18-kilodalton protein:Sanitization of nucleotide pool" Proc.Natl.Acad.Sci.,U.S.A.89. 11021-11025 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Akiko Shiraishi: "cDNA and genomic sequences for mouse O^6-methylguanine-DNA methyltransferase" Carcinogenesis. 13. 289-296 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Chue Ling-Ling: "Specific amino acid sequences required for O^6-methylguanine-DNA methyltransferase activity : Analysis of three residues at or near the methyl acceptor site" Carcinogenesis. 13. 837-843 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Zenca Matijasevic: "Release of N^2, 3-ethenoguanine from chloroacetaldehyde-treated DNA by Escherichia coli 3-methyladenine-DNA glycosylase II" Proc. Natl. Acad. Sci. U.S.A.89. 9331-9334 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Masato Furuichi: "Regulatory elements for expression of the alkA gene in response to alkylating agents" Molec. gen. Genet.236. 25-32 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Jin-Yao Mo: "Hydrolytic elimination of a mutagenic nucleotide, 8-oxodGTP, by human 18-kilodalton protein : Sanitization of nucleotide pool" Proc. Natl. Acad. Sci. U.S.A.89. 11021-11025 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 中鶴 陽子,松隈 章一,関口 睦夫,石川 隆俊: "Characterization of O^6ーmethylguanineーDNA methyltransferase in transgenic mice introduced with the E.coli ada gene" Mutation Res.254. 225-230 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Habraken,C.A.Carter,関口 睦夫,D.B.Ludlum.: "Release of N^2,3ーethanoguanine from haloethylnitrosoureaーtreated DNA by E.coli 3ーmethyladenine DNA glycosylase II" Carcinogenesis. 12. 1971-1973 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 作見 邦彦,白石 明子,早川 浩,関口 睦夫: "Cloning and expression of cDNA of rat O^6ーmethylguanine DNA methyltransferase" Nucleic Acids Res.19. 5597-5601 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 莫 錦尭,真木 寿治,関口 睦夫: "Mutational specificity of the dnaE173 mutator associated with a defect in the catalytic subunit of DNA polymerase III of Escherichia coli" J.Mol.Biol.222. 925-936 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 真木 寿治,関口 睦夫: "MutT protein specifically hydrolyzes a potent mutagenic substrate for DNA synthesis" Nature. 355. 273-275 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 福原 正生,早川 浩,作見 邦彦,関口 睦夫: "Induced synthesis of O^6ーmethylguanineーDNA methyltransferase in rat hepatoma cells exposed to DNAーdamaging agents" Jpn.J.Cancer Res.83. 72-77 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Takahashi: "Interaction of Ada protein with DNA examined by fluorescence anisotropy of the protein" Biochemistry. 29. 3431-3463 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Takahashi: "Effects fo vanillin and oーvanillin on induction of DNArepair networks:modulation of mutagenesis in Escherichia coli" Mutation Res.230. 127-134 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Hayakawa: "Expression and cloning of complementary DNA for a human enzyme that repairs O^bーmethylguanine in DNA" J.Mol.Biol. 213. 739-747 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] G.Koike: "Purification,structure,and bichemical properties of human O^bーmethylguaneーDNA methyltrasferse" J.Biol.Chem.265. 14754-14762 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] H.Akimaru: "Positive and negative regulation of transcription by a Cleavage product of Ada protein" J.Mol.Biol.216. 261-273 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] K.Sakumi: "Structures and functions of DNA glycosylases" Mutation Res.236. 161-172 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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