Abstract
Burkholderia pseudomallei, a motile and rod Gram-negative bacterium, is the causative agent of melioidosis. The bacterium is an intracellular pathogen and that motility is generally crucial for their survival in a natural environment and for systemic infection inside a host. We report here a role of B. pseudomallei polyphosphate kinase in virulence, such as an oxidative stress response, motilities and biofilm formation. The polyphosphate kinase (ppk) mutant is susceptible to hydrogen peroxide in an oxidative stress condition, unable to perform swimming, swarming motilities, and has lower density biofilm forming capacity than the wild-type strain. We also demonstrated that both polyphosphate kinase and motile flagella are essential and independently involved in biofilm formation. The B. pseudomallei flagellin (fliC) mutant and B. mallei, a nonmotile species, are shown to produce higher density biofilm formation than the ppk mutant, but less than wild type B. pseudomallei.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alexeyev, M.F. 1999. The pKNOCK series of broad-host-range mobilizable suicide vectors for gene knockout and targeted DNA insertion into the chromosome of Gram-negative bacteria. BioTechniques 26, 824–828.
Arkhipov, A., P.L. Freddolino, K. Imada, K. Namba, and K. Schulten. 2006. Coarse-grained molecular dynamics simulations of a rotating bacterial flagellum. Biophysical. J. 91, 4589–4597.
Brown, M.R.W. and A. Kornberg. 2008. The long and short of itpolyphosphate, ppk, and bacterial survival. Trends. Biochem. Sci. 33, 284–290.
Candon, H.L., B.J. Allan, C.D. Fraley, and E.C. Gaynor. 2007. Polyphosphate kinase 1 is a pathogenesis determinant in Campylobacter jejuni. J. Bacteriol. 189, 8099–8108.
Chua, K.L. 2003. Flagella are virulence determinants of Burkholderia pseudomallei. Infect. Immun. 71, 1622–1629.
DeShazer, D., P.J. Brett, R. Carlyon, and D.E. Woods. 1997. Mutagenesis of Burkholderia pseudomallei with Tn5-OT182: isolation of motility mutants and molecular characterization of the flagellin structural gene. J. Bacteriol. 179, 2116–2125.
Fraley, C.D., P.H. Rashid, S.S.K. Lee, R. Gottschalk, J. Harrison, P.J. Wood, M.R.W. Brown, and A. Kornberg. 2007. A polyphosphate kinase 1 (ppk1) mutant of Pseudomonas aerugenosa exhibits multiple ultra-structural and functional defects. Proc. Natl. Acad. Sci. USA 104, 3526–3531.
Godoy, D., G. Randle, A.J. Simpson, D.M. Aanensen, T.L. Pitt, R. Kinoshita, and B.G. Spratt. 2003. Multilocus sequence typing and evolutionary relationships among the causative agents of melioidosis and glanders, Burkholderia pseudomallei and Burkholderia mallei. J. Clin. Microbiol. 41, 2068–2079.
Klausen, M., A. Heydorn, P. Ragas, L. Lambertsen, A. Aaes-Jorgensen, S. Molin, and T. Tolker-Nielsen. 2003. Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants. Mol. Microbiol. 48, 1511–1524.
Kornberg, A., N.N. Rao, and D. Aulit-Riche. 1999. Inorganic polyphosphate: a molecule of many functions. Annu. Rev. Biochem. 68, 89–125.
Kovach, M.E., R.W. Phillips, P.H. Elzer, R.M. Roop, and K.M. Peterson. 1994. pBBR1MCS: a broad-host-range cloning vector. BioTechniques 16, 800–802.
Loprasert, S., W. Whangsuk, R. Sallabhan, and S. Mongkolsuk. 2004. DpsA protects the human pathogen Burkholderia pseudomallei against organic hydroperoxide. Arch. Microbiol. 182, 96–101.
Low, K.B. 1991. Conjugational methods for mapping with Hfr and Fprime strains. Methods Enzymol. 204, 43–62.
Ma, L., K.D. Jackson, R.M. Landry, M.R. Parsek, and D.J. Wozniak. 2006. Analysis of Pseudomonas aerugenosa conditional psl variants reveals roles for the psl polysaccharide in adhesion and maintaining biofilms structure postattachment. J. Bacteriol. 188, 8213–8221.
McLean, R.J.C., M.B. Barnes, M.K. Windham, M. Merchant, M.R.J. Forstner, and C. Fuqua. 2005. Cell-cell influences on bacterial community development in aquatic biofilms. Appl. Environ. Microbiol. 71, 8987–8990.
Nierman, W.C., D. DeShazer, H.S. Kim, H. Tettelin, K.E. Nelson, T. Feldblyum, and R.L. Ulrich et al. 2004. Structural flexibility in Burkholderia mallei genome. Proc. Natl. Acad. Sci. USA 101, 14246–14251.
O’Toole, G.A., L.A. Pratt, P.I. Watnick, D.K. Newman, V.B. Weaver, and R. Kolter. 1999. Genetic approaches to study biofilms. Methods Enzymol. 310, 91–109.
Pratt, L.A. and R. Kolter. 1998. Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol. Microbiol. 30, 285–293.
Prigent-Combaret, C., G. Prensier, T.T. Le Thi, O. Vidal, P. Lejeune, and C. Dorel. 2000. Developmental pathway for biofilm formation in curli-producing Escherichia coli strains: role of flagella, curli and colanic acid. Environ. Microbiol. 2, 450–464.
Rashid, M.H. and A. Kornberg. 2000. Inorganic polyphosphate is needed for swimming, swarming and twitching motilities of Pseudomonas aerugenosa. Proc. Natl. Acad. Sci. USA 97, 4885–4890.
Rashid, M.H., K. Rumbaugh, L. Passador, D.G. Davies, A.N. Hamood, B.H. Iglewski, and A. Kornberg. 2000. Polyphosphate kinase is essential for biofilm development, quorum sensing, and virulence of Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 97, 9639–9641.
Richard, M.I., S.L. Michell, and P.C.E. Oyston. 2008. An intracellularly inducible gene involved in virulence and polyphosphate production in Francisella. J. Med. Microbiol. 57, 1183–1192.
Ryder, C., M. Byrd, and D.J. Wozniak. 2007. Role of polysaccharides in Pseudomonas aerugenosa biofilm development. Cur. Opin. Microbiol. 10, 644–648.
Sambrook, J. and W.D. Russell. 2001. Molecular Cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, N.Y., USA.
Schell, M.A., L. Lipscomb, and D. DeShazer. 2008. Comparative gnomics and an insect model rapidly identify novel virulence genes of Burkholderia mallei. J. Bacteriol. 190, 2306–2313.
Takenaka, S., M. Iwaku, and E. Hoshino. 2001. Artificial Pseudomonas aeruginosa biofilms and confocal laser scanning microscopic analysis. J. Infect. Chemother. 7, 87–93.
Tanpiboonsak, S., A. Paemanee, S. Bunyarataphan, and S. Tungpradabkul. 2004. PCR-RFLP based differentiation of Burkholderia mallei and Burkholderia pseudomallei. Mol. Cell. Probes 18, 97–101.
Watnick, P.I. and R. Kolter. 1999. Steps in the development of a Vibrio cholerae El Tor biofilm. Mol. Microbiol. 34, 586–595.
White, N. 2003. Melioidosis. The Lancet 361, 1715–1722.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tunpiboonsak, S., Mongkolrob, R., Kitudomsub, K. et al. Role of a Burkholderia pseudomallei polyphosphate kinase in an oxidative stress response, motilities, and biofilm formation. J Microbiol. 48, 63–70 (2010). https://doi.org/10.1007/s12275-010-9138-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-010-9138-5