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Molecular analysis of the aidD6: : Mu dl (bla lac) fusion mutation of Escherichia coli K12

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Summary

In this report we present genetic and biochemical evidence indicating that the aidD6: : Mu dl (bla lac) fusion is an insertion of Mu dl (bla lac) into the alkB coding sequence. We describe the phenotypic effects resulting from this mutation and compare them with the effects of alkB22, alkA and ada mutations. We also constructed an alkA alkB double mutant and compared its phenotype with that of the single mutant strains. The observation that the methyl methanesulfonate (MMS) and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) resistance of the double mutant is approximately at the level predicted from the additive sensitivity of each of the single mutants suggests that these two gene products act in different pathways of DNA repair.

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References

  • Antonucci TK, Weng P, Rutter WJ (1989) Eukaryotic promoters drive gene expression in Escherichia coli. J Biol Chem 264:17656–17659

    Google Scholar 

  • Crisona NJ, Nowak JA, Nagaishi H, Clark AJ (1980) Transposon-mediated conjugational transmission of nonconjugative plasmids. J Bacteriol 142:701–713

    Google Scholar 

  • Evensen G, Seeberg E (1982) Adaptation to alkylation resistance involves the induction of a DNA glycosylase. Nature 296:773–775

    Google Scholar 

  • Groenen MAM, Timmers E, Van De Putte P (1985) DNA sequences at the ends of the genome of bacteriophage Mu essential for transposition. Proc Natl Acad Sci USA 82:2087–2091

    Google Scholar 

  • Hamblin MR, Potter BVL (1985) E. coli Ada regulatory protein repairs the Sp diastereoisomer of alkylated DNA. FEBS Lett 189:315–317

    Google Scholar 

  • Kahmann R (1983) Methylation regulates the expression of a DNA modification function encoded by bacteriophage Mu. Cold Spring Harbor Symp Quant Biol 43:639–646

    Google Scholar 

  • Karran P, Hjelmgren T, Lindahl T (1982) Induction of a DNA glycosylase for N-methyaated purines is part of the adaptive response to alkylating agents. Nature 296:770–773

    Google Scholar 

  • Kataoka H, Yamamoto Y, Sekiguchi M (1983) A new gene (alkB) of Escherichia coli that controls sensitivity to methyl methane sulfonate. J Bacteriol 153:1301–1307

    Google Scholar 

  • Kondo H, Nakabeppu Y, Kataoka H, Kuhara S, Kawabata S, Sekiguchi M (1986) Structure and expression of the alkB gene of Escherichia coli related to the repair of alkylated DNA. J Biol Chem 261:15772–15777

    Google Scholar 

  • Lawley PD (1974) Some chemical aspects of dose-response relationships in alkylation mutagenesis. Mutat Res 23:283–295

    Google Scholar 

  • LeMotte PK, Walker GC (1985) Induction and autoregulation of ada, a positively acting element regulating the response of Escherichia coli K-12 to methylating agents. J Bacteriol 161:888–895

    Google Scholar 

  • Lindahl T (1982) DNA repair enzymes. Annu Rev Biochem 51:6187

    Google Scholar 

  • Lindahl T, Sedgwick B, Sekiguchi M, Nakabeppu Y (1988) Regulation and expression of the adaptive response to alkylating agents. Annu Rev Biochem 57:133–157

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY

    Google Scholar 

  • Margison GP, Cooper DP, Brennand J (1985) Cloning of the E. coli 06-methylguanine and methylphosphotriester methyltransferase gene using a functional DNA repair assay. Nucleic Acids Res 13:1939–1952

    Google Scholar 

  • McCarthy TV, Lindahl T (1985) Methyl phosphotriesters in alkylated DNA are repaired by the Ada regulatory protein of E. coli. Nucleic Acids Res 13:2683–2698

    Google Scholar 

  • McCarthy TV, Karran P, Lindahl T (1984) Inducible repair of O-alkylated DNA pyrimidines in Escherichia coli. EMBO J 3:545–550

    Google Scholar 

  • Nakabeppu Y, Sekiguchi M (1986) Regulatory mechanisms for induction of synthesis of repair enzymes in response to alkylating agents: Ada protein acts as a transcriptional regulator. Proc Natl Acad Sci USA 83:6297–6301

    Google Scholar 

  • Nakabeppu Y, Miyata T, Kondo H, Iwanaga S, Sekiguchi M (1984) Structure and expression of the alkA gene of Escherichia coli involved in adaptive response to alkylating agents. J Biol Chem 259:13730–13736

    Google Scholar 

  • Nakabeppu Y, Kondo H, Kawabata SI, Iwanaga S, Sekiguchi M (1985) Purification and structure of the intact Ada regulatory protein of E. coli K-12, O6-methylguanine-DNA methyltransferase. J Biol Chem 260:7281–7288

    Google Scholar 

  • Poteete AR, Volkert MR (1988) Activation of the RecF recombination pathway in Escherichia coli by bacteriophage λ and P22-encoded functions. J Bacteriol 170:4370–4381

    Google Scholar 

  • Poteete AR, Fenton AC, Murphy KC (1988) Modulation of Escherichia coli RecBCD activity by the bacteriophage ;, Gam and P22 Abe functions. J Bacteriol 170:2012–2021

    Google Scholar 

  • Shevell DE, LeMotte PK, Walker GC (1988) Alteration of the carboxyl-terminal domain of Ada protein influences its inducibility, specificity, and strength as a transcriptional activator. J Bacteriol 170:5263–5271

    Google Scholar 

  • Takano K, Nakkabeppu Y, Sekiguchi M (1988) Functional sites of the Ada regulatory protein of Escherichia coli; analysis by amino acid substitutions. J Mol Biol 201:261–271

    Google Scholar 

  • Teo I, Sedgwick B, Kilpatrick MW, McCarthy TV, Lindahl T (1986) The intracellular signal for induction of resistance to alkylating agents in E. coli. Cell 45:315–324

    Google Scholar 

  • Volkert MR (1988) Adaptive response of Escherichia coli to alkylation damage. Environ Mol Mutagen 11:241–255

    Google Scholar 

  • Volkert MR (1989) Altered induction of the adaptive response to alkylation damage in Escherichia coli recF mutants. J Bacteriol 171:99–103

    Google Scholar 

  • Volkert MR, Nguyen DC (1984) Induction of specific Escherichia coli genes by sublethal treatments with alkylating agents. Proc Natl Acad Sci USA 81:4110–4114

    Google Scholar 

  • Volkert MR, Nguyen DC, Beard KC (1986) Escherichia coli gene induction by alkylation treatment. Genetics 112:11–26

    Google Scholar 

  • Warner HR, Demple BF, Deutsch WA, Kane CM, Linn S (1980) Apurinic/apyrimidinic endonucleases in repair of pyrimidine dimers and other lesions in DNA. Proc Natl Acad Sci USA 77:4602–4606

    Google Scholar 

  • Weinfeld M, Drake AF, Saunders JK, Paterson MC (1985) Stereospecific removal of methylphosphotriesters from DNA by an E. coli Ada extract. Nucleic Acids Res 13:7067–7077

    Google Scholar 

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Communicated by M. Sekiguchi

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Volkert, M.R., Hajec, L.I. Molecular analysis of the aidD6: : Mu dl (bla lac) fusion mutation of Escherichia coli K12. Molec. Gen. Genet. 229, 319–323 (1991). https://doi.org/10.1007/BF00272173

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  • DOI: https://doi.org/10.1007/BF00272173

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