Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Characterization of the Nickel Resistance Gene from Legionella pneumophila: Attenuation of Nickel Resistance by ppk (polyphosphate kinase) Disruption in Escherichia coli

  • Hahm, Dae-Hyun (Department of Oriental Medical Science, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Yeon, Mi-Jung (Department of Oriental Medical Science, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Ko, Whae-Min (Department of Molecular Biology, Institute of Oriental Medicine, College of Oriental Medicine, Kyung Hee University) ;
  • Lee, Eun-Jooh (Department of Oriental Medical Science, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Lee, Hye-Jung (Department of Oriental Medical Science, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Shim, In-Sop (Department of Oriental Medical Science, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Kim, Hong-Yeoul (Deaprtment of Molecular Biology, Institute of Oriental Medicine, College of Oriental Medicine, Kyung Hee Uniersity)
  • Published : 2002.02.01
Download PDF
⟨ Previous Next ⟩

Abstract

A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2＋$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

Keywords

References

  1. Mol. Microbiol. v.28 Metal ion homeostasis and intracellular parasitism Agranoff, D. D.;S. Krishna https://doi.org/10.1046/j.1365-2958.1998.00790.x
  2. Current Protocols in Molecular Biology(2nd ed.) Ausubel, F. M.;R. Brent;R. E. Kingston;D. D. Moore;J. G. Seidman;J. A. Smith;K. Struhl
  3. Infec. Immun. v.64 Allelic exchange mutagenesis of nixA in Helicobacter pylori results in reduced nickel transport and urease activity Bauerfeind, P.;R. M. Garner;H. L. Mobley
  4. Ecotoxicol. Environ. Safety v.45 Microbial resistance to metals in the environment Bruins, M. R.;S. Kapil;F. W. Oehme https://doi.org/10.1006/eesa.1999.1860
  5. Microbial Infec. v.2 Bacterial urease in infectious diseases Burner, A. B.;Y.-Y. M. Chen. https://doi.org/10.1016/S1286-4579(00)00312-9
  6. Arch. Microbiol. v.173 Nickel transport system in microorganisms Eitinger, T.;M.-A. Mandrand-Berthelot https://doi.org/10.1007/s002030050001
  7. Microb. Pathogenesis v.22 Mutants in the CtpA copper transporting P-type ATPase reduce virulence of Listeria monocytogenes Francis, M. S.;C. J. Thomas https://doi.org/10.1006/mpat.1996.0092
  8. J. Bacteriol. v.182 A gene cluster involved in metal homeostasis in the cyanobacterium Synechocystis sp. strain PCC6803 Garcia-Dominguez, M.;L. Lopez-Maury;F. J. Florencio;J. C. Reyes https://doi.org/10.1128/JB.182.6.1507-1514.2000
  9. J. Bacteriol. v.183 NreB from Achromobacter xylosoxidans 31A is a nickel-induced transporter conferring nickel resistance Grass, G.;B. Fan;B. P. Rosen;K. Lemke;H.-G. Schlegel;C. Rensing https://doi.org/10.1128/JB.183.9.2803-2807.2001
  10. J. Bacteriol. v.182 Regulation of the cnr cobalt and nickel resistance determinant from Ralstonia sp. strain CH34 Grass, G.;C. Grobe;D. H. Nies https://doi.org/10.1128/JB.182.5.1390-1398.2000
  11. J. Bacteriol. v.175 The Escherichia coli K-12 "wild types" W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels Jensen, K. F. https://doi.org/10.1128/jb.175.11.3401-3407.1993
  12. J. Microbiol. Biotechnol. v.9 Effect of AL072, a novel anti-Legionella antibiotic, on growth and cell morphology of Legionella pneumophila Kang, B. C.;J. H. Park;Y. S. Lee;J.-W. Suh;J.-H. Chang;C.-H. Lee
  13. Annu. Rev. Biochem. v.68 Inorganic polyphosphate: A molecule of many functions Kornberg, A.;N. N. Rao;D. Ault-Riche https://doi.org/10.1146/annurev.biochem.68.1.89
  14. J. Biol. Chem. v.272 Guanosine tetra- and pentaphosphate promote accumulation of inorganic polyphosphate in E. coli Kuroda, A.;H. Murphy;M. Cashel;A. Kornberg https://doi.org/10.1074/jbc.272.34.21240
  15. Acta Physiol. Scand. Suppl. v.163 Properties and functions of the P-type ion pumps cloned from Helicobacter pylori Melchers, K.;L. Herrmann;F. Mauch;D. Bayle;D. Heuermann;T. Weitzenegger;A. Schuhmacher;G. Aschs;R. Haas;G. Bode;K. Bensch;K. P. Schafer
  16. Microbiol. Mol. Biol. Rev. v.62 Major facilitator superfamily Pao, S. S.;I. T. Paulsen;M. H. saier, Jr.
  17. Gene v.179 Bacterial resistances to toxic metal ions - a review Silver, S. https://doi.org/10.1016/S0378-1119(96)00326-5
  18. J. Bacteriol. v.176 Combined nickel-cobalt-cadmium resistance encoded by the ncc locus of Alcaligenes xylosoxidans 31A Schmidt, T.;H. G. Schlegel https://doi.org/10.1128/jb.176.22.7045-7054.1994
  19. Appl. Environ. Microbiol. v.57 High-level nickel resistance in Alcaligenes xylosoxydans 31A and Alcaligenes eutrophus KTO2 Schmidt, T.;R.-D. Stoppel;H. G. Schlegel
  20. Biochemistry v.65 Inorganic polyphosphate kinase: Their novel biological functions and applications Shiba, T.;K. Tsutsumi;K. Ishige;T. Noguchi
  21. Appl. Environ. Microbiol. v.50 Effects of metals on Legionella pneumophila growth in drinking water plumbing systems States, S. J.;L. F. Conley;M. Ceraso;T. E. Stephenson;R. S. Wolford;R. M. Wadowsky;R. B. Yee
  22. Biochim. Biophys. Acta v.1493 Involvement of inorganic polyphosphate in expression of SOS genes Tsutsumi, K.;M. Munekata;T. Shiba https://doi.org/10.1016/S0167-4781(00)00165-2