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The Helicobacter pylori mutY homologue HP0142 is an antimutator gene that prevents specific C to A transversions

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Abstract

Extensive genetic variability resulting from a high mutation rate and frequent recombination is a characteristic of Helicobacter pylori. Its average mutation rate is 1 × 10−6, similar to that of Escherichia coli mutator strains. Few genes involved in DNA repair have been functionally characterized in H. pylori. In E. coli, the DNA glycosylase MutY is a part of the base excision repair system. The H. pylori mutY homolog HP0142 was analyzed in this study. HP0142 was disrupted by inserting a kanamycin resistance cassette. Mutation rates were determined by measuring the frequency of point mutations in rpoB conferring resistance against rifampicin. Inactivation of mutY in H. pylori resulted in an increase of the mutation frequency by a factor of up to 34. Sequence analysis of rpoB in rifampicin-resistant clones selected from the mutY mutant showed a modest increase of G:C/T:A transversions in comparison to clones selected from wild type strains. In contrast, inactivation of mutY had a profound impact on the distribution of mutations within rpoB. This finding suggests that the efficiency with which mutY prevents transversions is strongly dependent upon the sequence context. Inactivation of mutY was associated with a stationary phase fitness deficit in competitive cultures with the wild type strain.

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References

  • Achtman M, Azuma T, Berg DE, Ito Y, Morelli G, Pan ZJ, Suerbaum S, Thompson SA, van der Ende A, van Doorn LJ (1999) Recombination and clonal groupings within Helicobacter pylori from different geographic regions. Mol Microbiol 32:459–470

    Article  PubMed  CAS  Google Scholar 

  • Akopyanz N, Bukanov NO, Westblom TU, Kresovich S, Berg DE (1992) DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting. Nucleic Acids Res 20:5137–5142

    Article  PubMed  CAS  Google Scholar 

  • Alm RA, Ling LS, Moir DT, King BL, Brown ED, Doig PC, Smith DR, Noonan B, Guild BC, deJonge BL, Carmel G, Tummino PJ, Caruso A, Uria-Nickelsen M, Mills DM, Ives C, Gibson R, Merberg D, Mills SD, Jiang Q, Taylor DE, Vovis GF, Trust TJ (1999) Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature 397:176–180

    Article  PubMed  Google Scholar 

  • Au KG, Clark S, Miller JH, Modrich P (1989) Escherichia coli mutY gene encodes an adenine glycosylase active on G-A mispairs. Proc Natl Acad Sci USA 86:8877–8881

    Article  PubMed  CAS  Google Scholar 

  • Bjorkholm B, Sjolund M, Falk PG, Berg OG, Engstrand L, Andersson DI (2001) Mutation frequency and biological cost of antibiotic resistance in Helicobacter pylori. Proc Natl Acad Sci USA 98:14607–14612

    Article  PubMed  CAS  Google Scholar 

  • Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Collado-Vides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y (1997) The complete genome sequence of Escherichia coli K-12. Science 277:1453–1474

    Article  PubMed  CAS  Google Scholar 

  • David SS, Williams SD (1998) Chemistry of glycosylases and endonucleases involved in base-excision repair. Chem Rev 98:1221–1262

    Article  PubMed  CAS  Google Scholar 

  • Eisen JA (1998) A phylogenomic study of the MutS family of proteins. Nucleic Acids Res 26:4291–4300

    Article  PubMed  CAS  Google Scholar 

  • Eutsey R, Wang G, Maier RJ (2007) Role of a MutY DNA glycosylase in combating oxidative DNA damage in Helicobacter pylori. DNA Repair (Amst) 6:19–26

    Article  CAS  Google Scholar 

  • Falush D, Kraft C, Correa P, Taylor NS, Fox JG, Achtman M, Suerbaum S (2001) Recombination and mutation during long-term gastric colonization by Helicobacter pylori: Estimates of clock rates, recombination size and minimal age. Proc Natl Acad Sci USA 98:15056–15061

    Article  PubMed  CAS  Google Scholar 

  • Falush D, Wirth T, Linz B, Pritchard JK, Stephens M, Kidd M, Blaser MJ, Graham DY, Vacher S, Perez-Perez GI, Yamaoka Y, Megraud F, Otto K, Reichard U, Katzowitsch E, Wang X, Achtman M, Suerbaum S (2003) Traces of human migrations in Helicobacter pylori populations. Science 299:1582–1585

    Article  PubMed  CAS  Google Scholar 

  • Ferrero RL, Cussac V, Courcoux P, Labigne A (1992) Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol 174:4212–4217

    PubMed  CAS  Google Scholar 

  • Garibyan L, Huang T, Kim M, Wolff E, Nguyen A, Nguyen T, Diep A, Hu K, Iverson A, Yang H, Miller JH (2003) Use of the rpoB gene to determine the specificity of base substitution mutations on the Escherichia coli chromosome. DNA Repair (Amst) 2:593–608

    Article  CAS  Google Scholar 

  • Guan Y, Manuel RC, Arvai AS, Parikh SS, Mol CD, Miller JH, Lloyd S, Tainer JA (1998) MutY catalytic core, mutant and bound adenine structures define specificity for DNA repair enzyme superfamily. Nat Struct Biol 5:1058–1064

    Article  PubMed  CAS  Google Scholar 

  • Haas R, Meyer TF, van Putten JP (1993) Aflagellated mutants of Helicobacter pylori generated by genetic transformation of naturally competent strains using transposon shuttle mutagenesis. Mol Microbiol 8:753–760

    Article  PubMed  CAS  Google Scholar 

  • Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580

    Article  PubMed  CAS  Google Scholar 

  • Heep M, Beck D, Bayerdorffer E, Lehn N (1999) Rifampin and rifabutin resistance mechanism in Helicobacter pylori. Antimicrob Agents Chemother 43:1497–1499

    PubMed  CAS  Google Scholar 

  • Huang S, Kang J, Blaser MJ (2006) Antimutator role of the DNA glycosylase mutY gene in Helicobacter pylori. J Bacteriol 188:6224–6234

    Article  PubMed  CAS  Google Scholar 

  • Kansau I, Raymond J, Bingen E, Courcoux P, Kalach N, Bergeret M, Braimi N, Dupont C, Labigne A (1996) Genotyping of Helicobacter pylori isolates by sequencing of PCR products and comparison with the RAPD technique. Res Microbiol 147:661–669

    Article  PubMed  CAS  Google Scholar 

  • Kim M, Huang T, Miller JH (2003) Competition between MutY and mismatch repair at A × C mispairs in vivo. J Bacteriol 185:4626–4629

    Article  PubMed  CAS  Google Scholar 

  • Labigne-Roussel A, Courcoux P, Tompkins L (1988) Gene disruption and replacement as a feasible approach for mutagenesis of Campylobacter jejuni. J Bacteriol 170:1704–1708

    PubMed  CAS  Google Scholar 

  • Langenberg W, Rauws EA, Widjojokusumo A, Tytgat GN, Zanen HC (1986) Identification of Campylobacter pyloridis isolates by restriction endonuclease DNA analysis. J Clin Microbiol 24:414–417

    PubMed  CAS  Google Scholar 

  • Linz B, Balloux F, Moodley Y, Hua L, Manica A, Roumagnac P, Falush D, Stamer C, Prugnolle F, van der Merwe SW, Yamaoka Y, Graham DY, Perez-Trallero E, Wadstrom T, Suerbaum S, Achtman M (2007) An African origin for the intimate association between humans and Helicobacter pylori. Nature 445:915–918

    Article  PubMed  Google Scholar 

  • Majewski SI, Goodwin CS (1988) Restriction endonuclease analysis of the genome of Campylobacter pylori with a rapid extraction method: evidence for considerable genomic variation. J Infect Dis 157:465–471

    PubMed  CAS  Google Scholar 

  • Mathieu A, O’Rourke EJ, Radicella JP (2006) Helicobacter pylori genes involved in avoidance of mutations induced by 8-oxoguanine. J Bacteriol 188:7464–7469

    Article  PubMed  CAS  Google Scholar 

  • Pinto AV, Mathieu A, Marsin S, Veaute X, Ielpi L, Labigne A, Radicella JP (2005) Suppression of homologous and homeologous recombination by the bacterial MutS2 protein. Mol Cell 17:113–120

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Russell DG (2004) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Sanchez AM, Volk DE, Gorenstein DG, Lloyd RS (2003) Initiation of repair of A/G mismatches is modulated by sequence context. DNA Repair (Amst) 2:863–878

    Article  CAS  Google Scholar 

  • Schmitz A, Josenhans C, Suerbaum S (1997) Cloning and characterization of the Helicobacter pylori flbA gene, which codes for a membrane protein involved in coordinated expression of flagellar genes. J Bacteriol 179:987–997

    PubMed  CAS  Google Scholar 

  • Suerbaum S, Josenhans C (2007) Helicobacter pylori evolution and phenotypic diversification in a changing host. Nat Rev Microbiol 5:441–452

    Article  PubMed  CAS  Google Scholar 

  • Suerbaum S, Michetti P (2002) Helicobacter pylori infection. N Engl J Med 347:1175–1186

    Article  PubMed  CAS  Google Scholar 

  • Tomb J-F, White O, Kerlavage AR, Clayton RA, Sutton GG, Fleischmann RD, Ketchum KA, Klenk HP, Gill S, Dougherty BA, Nelson K, Quackenbush J, Zhou L, Kirkness EF, Peterson S, Loftus B, Richardson D, Dodson R, Khalak HG, Glodek A, McKenney K, Fitzegerald LM, Lee N, Adams MD, Hickey EK, Berg DE, Gocayne JD, Utterback TR, Peterson JD, Kelley JM, Cotton MD, Weidman JM, Fujii C, Bowman C, Watthey L, Wallin E, Hayes WS, Borodovsky M, Karp PD, Smith HO, Fraser CM, Venter JC (1997) The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 388:539–547

    Article  PubMed  CAS  Google Scholar 

  • Tsai-Wu JJ, Liu HF, Lu AL (1992) Escherichia coli MutY protein has both N-glycosylase and apurinic/apyrimidinic endonuclease activities on A.C and A.G mispairs. Proc Natl Acad Sci USA 89:8779–8783

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank Schalk van der Merwe for permission to use strain SA166A, Bodo Linz and Mark Achtman for permission to use strain Ala15, and Christine Josenhans for fruitful discussions and critical reading of the paper. This work was supported by funding under the Sixth Research Framework Programme of the European Union, project INCA (LSHC-CT-2005–018704). S.K. received a Ph.D. stipend from Research Training Group GRK745 funded by the German Research Foundation (DFG). C.M. received a Ph.D. stipend from the German Academic Exchange Service (DAAD) and the Wilhelm-Hirte Foundation.

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Author notes

  1. Christian Kraft

    Present address: University of Massachusetts Medical School, Program in Gene Function and Expression, 364 Plantation Street, Worcester, MA, 01605, USA

Authors and Affiliations

  1. Department of Medical Microbiology and Hospital Epidemiology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    Stefan Kulick, Claudia Moccia, Christian Kraft & Sebastian Suerbaum

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  1. Stefan Kulick
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  2. Claudia Moccia
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  4. Sebastian Suerbaum
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Correspondence to Sebastian Suerbaum.

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Communicated by Erko Stackebrandt.

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Kulick, S., Moccia, C., Kraft, C. et al. The Helicobacter pylori mutY homologue HP0142 is an antimutator gene that prevents specific C to A transversions. Arch Microbiol 189, 263–270 (2008). https://doi.org/10.1007/s00203-007-0315-9

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  • DOI: https://doi.org/10.1007/s00203-007-0315-9

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