Abstract
Infections are often not caused by a colonization of Pseudomonas aeruginosa alone but by a consortium of other bacteria. Little is known about the impact of P. aeruginosa on the growth of other bacteria upon coinfection. Here, cellree culture supernatants obtained from P. aeruginosa suppressed the growth of a number of bacterial strains such as Corynebacterium glutamicum, Bacillus subtilis, Staphylococcus aureus, and Agrobacterium tumefaciens, but had little effect on the growth of Escherichia coli and Salmonella Typhimurium. The growth suppression effect was obvious when P. aeruginosa was cultivated in M9 minimal media, and the suppression was not due to pyocyanin, a well-known antimicrobial toxin secreted by P. aeruginosa. By performing transposon mutagenesis, PA5070 encoding TatC was identified, and the culture supernatant of its mutant did not suppress the growth. HPLC analysis of supernatants showed that pyoverdine was a secondary metabolite present in culture supernatants of the wild-type strain, but not in those of the PA5070 mutant. Supplementation of FeCl2 as a source of iron compromised the growth suppression effect of supernatants and also recovered biofilm formation of S. aureus, indicating that pyoverdine-mediated iron acquisition is responsible for the growth suppression. Thus, this study provides the action of TatC-dependent pyoverdine translocation for the growth suppression of other bacteria, and it might aid understanding of the impact of P. aeruginosa in the complex community of bacterial species upon coinfection.
Similar content being viewed by others
References
Arima, K., Fukuta, A., Imanaka, H., Kousaka, M., and Tamura, G. 1964. Pyrrolnitrin new antibiotic substance produced by Pseudomonas. Agr. Biol. Chem. Tokyo 28, 575–576.
Biswas, L., Biswas, R., Nerz, C., Ohlsen, K., Schlag, M., Schafer, T., Lamkemeyer, T., Ziebandt, A.K., Hantke, K., Rosenstein, R., et al. 2009. Role of the twin-arginine translocation pathway in Staphylococcus. J. Bacteriol. 191, 5921–5929.
Caetano-Anolles, G. 1993. Amplifying DNA with arbitrary oligonucleotide primers. PCR Methods Appl. 3, 85–94.
Carrano, C.J. and Raymond, K.N. 1979. Ferric ion sequestering agents. 2. Kinetics and mechanism of iron removal from transferrin by enterobactin and synthetic tricatechols. J. Am. Chem. Soc. 101, 5401–5404.
Ding, Z. and Christie, P.J. 2003. Agrobacterium tumefaciens twinarginine-dependent translocation is important for virulence, flagellation, and chemotaxis but not type iv secretion. J. Bacteriol. 185, 760–771.
Folkesson, A., Jelsbak, L., Yang, L., Johansen, H.K., Ciofu, O., Hoiby, N., and Molin, S. 2012. Adaptation of Pseudomonas aeruginosa to the cystic fibrosis airway: An evolutionary perspective. Nat. Rev. Microbiol. 10, 841–851.
Follettie, M.T., Peoples, O.P., Agoropoulou, C., and Sinskey, A.J. 1993. Gene structure and expression of the Corynebacterium flavum N13 ask-asd operon. J. Bacteriol. 175, 4096–4103.
Handfield, M., Lehoux, D.E., Sanschagrin, F., Mahan, M.J., Woods, D.E., and Levesque, R.C. 2000. In vivo-induced genes in Pseudomonas aeruginosa. Infect. Immun. 68, 2359–2362.
Hassan, H.M. and Fridovich, I. 1980. Mechanism of the antibiotic action pyocyanine. J. Bacteriol. 141, 156–163.
Holloway, B.W., Krishnapillai, V., and Morgan, A.F. 1979. Chromosomal genetics of Pseudomonas. Microbiol. Rev. 43, 73–102.
Howell, C.R. and Stipanovic, R.D. 1980. Suppression of Pythium ultimum-induced damping-off of cotton seedlings by Pseudomonas fluorescens and its antibiotic, pyoluteorin. Phytopathology 70, 712–715.
Hunt, T.A., Peng, W.T., Loubens, I., and Storey, D.G. 2002. The Pseudomonas aeruginosa alternative sigma factor PvdS controls exotoxin A expression and is expressed in lung infections associated with cystic fibrosis. Microbiology 148, 3183–3193.
Kessler, E., Safrin, M., Olson, J.C., and Ohman, D.E. 1993. Secreted LasA of Pseudomonas aeruginosa is a staphylolytic protease. J. Biol. Chem. 268, 7503–7508.
Kiernan, L., Finkelstein, F.O., Kliger, A.S., Gorban-Brennan, N., Juergensen, P., Mooraki, A., and Brown, E. 1995. Outcome of polymicrobial peritonitis in continuous ambulatory peritoneal dialysis patients. Am. J. Kidney Dis. 25, 461–464.
Kikuchi, Y., Date, M., Itaya, H., Matsui, K., and Wu, L.F. 2006. Functional analysis of the twin-arginine translocation pathway in Corynebacterium glutamicum ATCC 13869. Appl. Environ. Microbiol. 72, 7183–7192.
Kluge, S., Hoffmann, M., Benndorf, D., Rapp, E., and Reichl, U. 2012. Proteomic tracking and analysis of a bacterial mixed culture. Proteomics 12, 1893–1901.
Kulasekara, H.D., Ventre, I., Kulasekara, B.R., Lazdunski, A., Filloux, A., and Lory, S. 2005. A novel two-component system controls the expression of Pseudomonas aeruginosa fimbrial cup genes. Mol. Microbiol. 55, 368–380.
Lamont, I.L., Beare, P.A., Ochsner, U., Vasil, A.I., and Vasil, M.L. 2002. Siderophore-mediated signaling regulates virulence factor production in Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 99, 7072–7077.
Linget, C., Stylianou, D.G., Dell, A., Wolff, R.E., Piemont, Y., and Abdallah, M.A. 1992. Bacterial siderophores - the structure of a desferriferribactin produced by Pseudomonas fluorescens ATCC 13525. Tetrahedron Lett. 33, 3851–3854.
Liu, P.V. 1966. The roles of various fractions of Pseudomonas aeruginosa in its pathogenesis. 3. Identity of the lethal toxins produced in vitro and in vivo. J. Infect. Dis. 116, 481–489.
Meyer, J.M., Neely, A., Stintzi, A., Georges, C., and Holder, I.A. 1996. Pyoverdin is essential for virulence of Pseudomonas aeruginosa. Infect. Immun. 64, 518–523.
Muller, M. 2005. Twin-arginine-specific protein export in Escherichia coli. Res. Microbiol. 156, 131–136.
Nielsen, T.H., Christophersen, C., Anthoni, U., and Sorensen, J. 1999. Viscosinamide, a new cyclic depsipeptide with surfactant and antifungal properties produced by Pseudomonas fluorescens DR54. J. Appl. Microbiol. 87, 80–90.
Nielsen, T.H., Thrane, C., Christophersen, C., Anthoni, U., and Sorensen, J. 2000. Structure, production characteristics and fungal antagonism of tensin - a new antifungal cyclic lipopeptide from Pseudomonas fluorescens strain 96.578. J. Appl. Microbiol. 89, 992–1001.
Ochsner, U.A., Snyder, A., Vasil, A.I., and Vasil, M.L. 2002. Effects of the twin-arginine translocase on secretion of virulence factors, stress response, and pathogenesis. Proc. Natl. Acad. Sci. USA 99, 8312–8317.
Oertel, D., Schmitz, S., and Freudl, R. 2015. A tatabc-type tat translocase is required for unimpaired aerobic growth of Corynebacterium glutamicum ATCC13032. PLoS One 10, e0123413.
O’Toole, G.A. and Kolter, R. 1998. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol. Microbiol. 30, 295–304.
Otto, B.R., Verweij-van Vught, A.M., and MacLaren, D.M. 1992. Transferrins and heme-compounds as iron sources for pathogenic bacteria. Crit. Rev. Microbiol. 18, 217–233.
Pernet, E., Guillemot, L., Burgel, P.R., Martin, C., Lambeau, G., Sermet-Gaudelus, I., Sands, D., Leduc, D., Morand, P.C., Jeammet, L., et al. 2014. Pseudomonas aeruginosa eradicates Staphylococcus aureus by manipulating the host immunity. Nat. Commun. 5, 5105.
Rahme, L.G., Stevens, E.J., Wolfort, S.F., Shao, J., Tompkins, R.G., and Ausubel, F.M. 1995. Common virulence factors for bacterial pathogenicity in plants and animals. Science 268, 1899–1902.
Ramos, I., Dietrich, L.E., Price-Whelan, A., and Newman, D.K. 2010. Phenazines affect biofilm formation by Pseudomonas aeruginosa in similar ways at various scales. Res. Microbiol. 161, 187–191.
Razvi, S., Quittell, L., Sewall, A., Quinton, H., Marshall, B., and Saiman, L. 2009. Respiratory microbiology of patients with cystic fibrosis in the united states, 1995 to 2005. Chest 136, 1554–1560.
Reynolds, M.M., Bogomolnaya, L., Guo, J., Aldrich, L., Bokhari, D., Santiviago, C.A., McClelland, M., and Andrews-Polymenis, H. 2011. Abrogation of the twin arginine transport system in Salmonella enterica serovar Typhimurium leads to colonization defects during infection. PLoS One 6, e15800.
Ruger, M., Ackermann, M., and Reichl, U. 2014. Species-specific viability analysis of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus in mixed culture by flow cytometry. BMC Microbiol. 14, 56.
Settles, A.M., Yonetani, A., Baron, A., Bush, D.R., Cline, K., and Martienssen, R. 1997. Sec-independent protein translocation by the maize Hcf106 protein. Science 278, 1467–1470.
Shin, H.S., Kim, Y.J., Yoo, I.H., Lee, H.S., Jin, S., and Ha, U.H. 2011. Autoinduction of a genetic locus encoding putative acyltransferase in Corynebacterium glutamicum. Biotechnol. Lett. 33, 97–102.
Simone, D., Bay, D.C., Leach, T., and Turner, R.J. 2013. Diversity and evolution of bacterial twin arginine translocase protein, tatc, reveals a protein secretion system that is evolving to fit its environmental niche. PLoS One 8, e78742.
Slininger, P.J., Burkhead, K.D., Schisler, D.A., and Bothast, R.J. 2000. Isolation, identification, and accumulation of 2-acetamidophenol in liquid cultures of the wheat take-all biocontrol agent Pseudomonas fluorescens 2-79. Appl. Microbiol. Biotechnol. 54, 376–381.
Snyder, A., Vasil, A.I., Zajdowicz, S.L., Wilson, Z.R., and Vasil, M.L. 2006. Role of the Pseudomonas aeruginosa PlcH Tat signal peptide in protein secretion, transcription, and cross-species Tat secretion system compatibility. J. Bacteriol. 188, 1762–1774.
Straight, P.D. and Kolter, R. 2009. Interspecies chemical communication in bacterial development. Annu. Rev. Microbiol. 63, 99–118.
Takase, H., Nitanai, H., Hoshino, K., and Otani, T. 2000. Impact of siderophore production on Pseudomonas aeruginosa infections in immunosuppressed mice. Infect. Immun. 68, 1834–1839.
Torres, L., Perez-Ortin, J.E., Tordera, V., and Beltran, J.P. 1986. Isolation and characterization of an Fe(iii)-chelating compound produced by Pseudomonas syringae. Appl. Environ. Microbiol. 52, 157–160.
Watson, D., MacDermot, J., Wilson, R., Cole, P.J., and Taylor, G.W. 1986. Purification and structural analysis of pyocyanin and 1-hydroxyphenazine. Eur. J. Biochem. 159, 309–313.
Weaver, V.B. and Kolter, R. 2004. Burkholderia spp. alter Pseudomonas aeruginosa physiology through iron sequestration. J. Bacteriol. 186, 2376–2384.
West, S.E., Schweizer, H.P., Dall, C., Sample, A.K., and Runyen-Janecky, L.J. 1994. Construction of improved Escherichia-Pseudomonas shuttle vectors derived from pUC18/19 and sequence of the region required for their replication in Pseudomonas aeruginosa. Gene 148, 81–86.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, Y., Kim, YJ., Lee, JH. et al. TatC-dependent translocation of pyoverdine is responsible for the microbial growth suppression. J Microbiol. 54, 122–130 (2016). https://doi.org/10.1007/s12275-016-5542-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-016-5542-9

