DOI QR코드

DOI QR Code

Expression of the mexA Gene Requires the DNA Helicase RecG in Pseudomonas aeruginosa PAO1

  • Heo, Aram (Laboratory of Molecular Environmental Microbiology, Department of Environmental Science and Ecological Engineering, Korea University) ;
  • Park, Woojun (Laboratory of Molecular Environmental Microbiology, Department of Environmental Science and Ecological Engineering, Korea University)
  • Received : 2014.10.06
  • Accepted : 2014.11.10
  • Published : 2015.04.28

Abstract

This study provides evidence that RecG regulates the expression of the OxyR-independent gene mexA in Pseudomonas aeruginosa PAO1. A reduction in mexA expression was observed in the absence of RecG, but not OxyR, by northern blot and quantitative real-time PCR analyses. The canonical palindromic RecG binding sequence was present upstream of the mexA promoter, and bound purified RecG and single strand-binding protein. These data reveal a novel mechanism of OxyR-independent gene transcription by RecG.

Keywords

References

  1. Buss JA, Kimura Y, Bianco PR. 2008. RecG interacts directly with SSB: implications for stalled replication fork regression. Nucleic Acids Res. 36: 7029-7042. https://doi.org/10.1093/nar/gkn795
  2. Buyck JM, Guénard S, Plésiat P, Tulkens PM, Van Bambeke F. 2012. Role of MexAB-OprM in intrinsic resistance of Pseudomonas aeruginosa to temocillin and impact on the susceptibility of strains isolated from patients suffering from cystic fibrosis. J. Antimicrob. Chemother. 67: 771-775. https://doi.org/10.1093/jac/dkr543
  3. Cañas C, Suzuki Y, Marchisone C, Carrasco B, Freire-Benéitez V, Takeyasu K, et al. 2014. Interaction of branch migration translocases with the Holliday junction-resolving enzyme and their implications in Holliday junction resolution. J. Biol. Chem. 289: 17634-17646. https://doi.org/10.1074/jbc.M114.552794
  4. Daigle DM, Cao L, Fraud S, Wilke MS, Pacey A, Klinoski R, et al. 2007. Protein modulator of multidrug efflux gene expression in Pseudomonas aeruginosa. J. Bacteriol. 189: 5441-5451. https://doi.org/10.1128/JB.00543-07
  5. Evans K, Adewoye L, Poole K. 2001. MexR repressor of the mexAB-oprM multidrug efflux operon of Pseudomonas aeruginosa: identification of MexR binding sites in the mexA-mexR intergenic region. J. Bacteriol. 183: 807-812. https://doi.org/10.1128/JB.183.3.807-812.2001
  6. Li XZ, Nikaido H, Poole K. 1995. Role of mexA-mexB-oprM in antibiotic efflux in Pseudomonas aeruginosa. Antimicrob. Agents. Chemother. 39: 1948-1953. https://doi.org/10.1128/AAC.39.9.1948
  7. Lloyd RG, Buckman C. 1991. Genetic analysis of the recG locus of Escherichia coli K-12 and of its role in recombination and DNA repair. J. Bacteriol. 173: 1004-1011. https://doi.org/10.1128/jb.173.3.1004-1011.1991
  8. Maseda H, Yoneyama H, Nakae T. 2000. Assignment of the substrate-selective subunits of the MexEF-OprN multidrug efflux pump of Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 44: 658-664. https://doi.org/10.1128/AAC.44.3.658-664.2000
  9. McGlynn P, Mahdi AA, Lloyd RG. 2000. Characterisation of the catalytically active form of RecG helicase. Nucleic Acids Res. 28: 2324-2332. https://doi.org/10.1093/nar/28.12.2324
  10. Morita Y, Cao L, Gould G, Avison MB, Poole K. 2006. nalD encodes a second repressor of the mexAB-oprM multidrug efflux operon of Pseudomonas aeruginosa. J. Bacteriol. 188: 8649-8654. https://doi.org/10.1128/JB.01342-06
  11. Ochsner UA, Vasil ML, Alsabbagh E, Parvatiyar K, Hassett DJ. 2000. Role of the Pseudomonas aeruginosa oxyR-recG operon in oxidative stress defense and DNA repair: OxyR-dependent regulation of katB-ankB, ahpB, and ahpC-ahpF. J. Bacteriol. 182: 4533-4544. https://doi.org/10.1128/JB.182.16.4533-4544.2000
  12. Ochsner UA, Hassett DJ, Vasil ML. 2001. Genetic and physiological characterization of ohr, encoding a protein involved in organic hydroperoxide resistance in Pseudomonas aeruginosa. J. Bacteriol. 183: 773-778. https://doi.org/10.1128/JB.183.2.773-778.2001
  13. Poole K, Srikumar R. 2001. Multidrug efflux in Pseudomonas aeruginosa: components, mechanisms and clinical significance. Curr. Top. Med. Chem. 1: 59-71. https://doi.org/10.2174/1568026013395605
  14. Sánchez P, Rojo F, Martínez JL. 2002. Transcriptional regulation of mexR, the repressor of Pseudomonas aeruginosa mexAB-oprM multidrug efflux pump. FEMS Microbiol. Lett. 207: 63-68. https://doi.org/10.1111/j.1574-6968.2002.tb11029.x
  15. Sharples GJ, Ingleston SM, Lloyd RG. 1999. Holliday junction processing in bacteria: insights from the evolutionary conservation of RuvABC, RecG, and RusA. J. Bacteriol. 181: 5543-5550.
  16. Yeom J, Lee Y, Park W. 2012. ATP-dependent RecG helicase is required for the transcriptional regulator OxyR function in Pseudomonas species. J. Biol. Chem. 287: 24492-24504. https://doi.org/10.1074/jbc.M112.356964