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A mutation in tdcA attenuates the virulence of Salmonella enterica serovar Typhimurium

Molecules and Cells volume 29, pages 509–517 (2010)Cite this article

Abstract

The Salmonella tdc operon encodes enzymes belonging to a metabolic pathway that degrades l-serine and l-threonine. The upregulation of the tdc operon and increased virulence of Salmonella grown under oxygenlimiting conditions prompted us to investigate the role of the tdc operon in the pathogenesis of Salmonella Typhimurium. A Salmonella strain carrying a null mutation in tdcA, which encodes the transcriptional activator of the tdc operon, was impaired in mice infected intraperitoneally with the bacterium. In addition, the Salmonella tdcA mutant showed reduced replication compared with the parental strain in cultured animal cells, although their growth rates were similar in various culture media. To understand the function of TdcA in pathogenesis, we performed two-dimensional gel electrophoresis and found that flagellar and PhoP-regulated proteins were affected by the tdcA mutation. The results of β-galactosidase assays and FACS analysis showed that, among the four PhoP-dependent genes tested, the expression of ssaG, which is located in Salmonella pathogenicity island 2 (SPI2), was reduced in the tdcA mutant, especially in the intracellular environment of macrophages. Taken together, our data suggest that tdcA plays an important role in the pathogenesis of Salmonella.

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References

  • Altier, C. (2005). Genetic and environmental control of Salmonella invasion. J. Microbiol. 43, 85–92.

    CAS  PubMed  Google Scholar 

  • Becker, D., Selbach, M., Rollenhagen, C., Ballmaier, M., Meyer, T.F., Mann, M., and Bumann, D. (2006). Robust Salmonella metabolism limits possibilities for new antimicrobials. Nature 440, 303–307.

    Article  CAS  PubMed  Google Scholar 

  • Behlau, I., and Miller, S.I. (1995). A phoP-repressed gene promotes Salmonella typhimurium invasion of epithelial cells. J. Bacteriol. 175, 4475–4484.

    Google Scholar 

  • Bijisma, J.J.E., and Groisman, E.A. (2005). The PhoP/PhoQ system controls the intramacrophage type three secretion system of Salmonella enterica. Mol. Microbiol. 57, 85–96.

    Article  Google Scholar 

  • Cirillo, D.M., Valdivia, R.H., Monack, D.M., and Falkow, S. (1998). Macrophage-dependent induction of the Salmonella pathogenicity island 2 type III secretion system and its role in intracellular survival. Mol. Micriobiol. 30, 175–188.

    Article  CAS  Google Scholar 

  • Datsenko, K.A., and Wanner, B.L. (2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. USA. 97, 6640–6645.

    Article  CAS  PubMed  Google Scholar 

  • Deiwick, J., Rappl, C., Stender, S., Jungblut, P.R., and Hensel, M. (2002). Proteomic approaches to Salmonella pathogenicity island 2 encoded proteins and the SsrAB regulon. Proteomics 2, 792–799.

    Article  CAS  PubMed  Google Scholar 

  • Eriksson, S., Lucchini, S., Thompson, A., Rhen, M., and Hinton, J.C. (2003). Unravelling the biology of macrophage infection by gene expression profiling of intracellular Salmonella enterica. Mol. Micriobiol. 47, 103–118.

    Article  CAS  Google Scholar 

  • Feng, X., Oropeza, R., and Kenney, L.J. (2003). Dual regulation by phospho-OmpR of ssrA/B gene expression in Salmonella pathogenicity island 2. Mol. Micriobiol. 48, 1131–1143.

    Article  CAS  Google Scholar 

  • Galán, J.E. (1996). Molecular genetic bases of Salmonella entry into host cells. Mol. Microbiol. 20, 263–271.

    Article  PubMed  Google Scholar 

  • Ganduri, Y.L., Sadda, S.R., Datta, M.W., Jambukeswaran, R.K., and Datta, P. (1993). TdcA, a transcriptional activator of the tdcABC operon of Escherichia coli, is a member of the LysR family of proteins. Mol. Gen. Genet. 240, 395–402

    CAS  PubMed  Google Scholar 

  • Goss, T.J., Schweizer, H.P., and Datta, P. (1988). Molecular characterization of the tdc operon of Escherichia coli K-12. J. Bacteriol. 170, 5352–5359.

    CAS  PubMed  Google Scholar 

  • Groisman, E.A. (2001). The pleiotropic two-component regulatory system PhoP-PhoQ. J. Bacteriol. 183, 1835–1842.

    Article  CAS  PubMed  Google Scholar 

  • Groisman, E.A., and Mouslim, C. (2006). Sensing by bacterial regulatory systems in host and non-host environments. Nat. Rev. Microbiol. 4, 705–709.

    Article  CAS  PubMed  Google Scholar 

  • Hautefort, I., Proença, M.J., and Hinton, J.C.D. (2003). Single-copy green fluorescent protein gene fusions allow accurate measurement of Salmonella gene expression in vitro and during infection of mammalian cells. Appl. Environ. Microbiol. 69, 7480–7491.

    Article  CAS  PubMed  Google Scholar 

  • Hensel, M. (2000). Salmonella pathogenicity island 2. Mol. Microbiol. 36, 1015–1023.

    Article  CAS  PubMed  Google Scholar 

  • Heßlinger, C., Fairhurst, S.A., and Sawers, G. (1998). Novel keto acid formate-lyase and propionate kinase enzymes are components of an anaerobic pathway in Escherichia coli that degrades l-threonine to propionate. Mol. Microbiol. 27, 477–492.

    Article  PubMed  Google Scholar 

  • Jones, B.D., and Falkow, S. (1996). Salmonellosis: host immune responses and bacterial virulence determinants. Annu. Rev. Immunol. 14, 533–561.

    Article  CAS  PubMed  Google Scholar 

  • Josenhans, C., and Suerbaum, S. (2002). The role of motility as a virulence factor in bacteria. Int. J. Med. Microbiol. 291, 605–614.

    Article  CAS  PubMed  Google Scholar 

  • Kier, L.D., Weppelman, R., and Ames, B.N. (1977). Regulation of two phosphatases and a cyclic phosphodiesterase of Salmonella typhimurium. J. Bacteriol. 130, 420–428.

    CAS  PubMed  Google Scholar 

  • Kim, C.C., and Falkow, S. (2004). Delineation of upstream signaling events in the Salmonella pathogenicity island 2 transcriptional activation pathway. J. Bacteriol. 186, 4694–4704.

    Article  CAS  PubMed  Google Scholar 

  • Kim, M., Lim, S., and Ryu, S. (2008). Molecular analysis of the Salmonella Typhimurium tdc operon regulation. J. Microbiol. Biotechnol. 18, 1024–1032.

    CAS  PubMed  Google Scholar 

  • Kim, M., Lim, S., Kim, D., Choy, H.E., and Ryu, S. (2009). A tdcA mutation reduces the invasive ability of Salmonella enterica serovar Typhimurium. Mol. Cells 28, 389–395.

    Article  CAS  PubMed  Google Scholar 

  • Lee, C.A., and Falkow, S. (1990). The ability of Salmonella to enter mammalian cells is affected by bacterial growth state. Proc. Natl. Acad. Sci. USA 87, 4304–4308.

    Article  CAS  PubMed  Google Scholar 

  • Lim, S., Kim, B., Choi, H.-S., Lee, Y., and Ryu, S. (2006). Fis is required for proper regulation of ssaG expression in Salmonella enterica serovar Typhimurium. Microb. Pathog. 41, 33–42.

    Article  CAS  PubMed  Google Scholar 

  • Linehan, S.A., Rytkönen, A., Yu, X.-J., Liu, M., and Holden, D.W. (2005). SlyA regulates function of Salmonella pathogenicity island 2 (SPI-2) and expression SPI-2-associated genes. Infect. Immun. 73, 4354–4362.

    Article  CAS  PubMed  Google Scholar 

  • Löner, S., Jäckel, D., Kaiser, N., and Hensel, M. (2006). Regulation of Salmonella pathogenicity island 2 genes by independent environmental signals. Int. J. Med. Microbiol. 296, 435–447.

    Article  Google Scholar 

  • Lucas, R.L., and Lee, C.A. (2000). Unraveling the mysteries of virulence gene regulation in Salmonella typhimurium. Mol. Microbiol. 36, 1024–1033.

    Article  CAS  PubMed  Google Scholar 

  • Maloy, S.R., Stewart, V.J., and Taylor, R.K. (1996). Genetic analysis of pathogenic bacteria: A Laboratory Manual, (Plainview, NY: Cold Spring Harbor Laboratory Press).

    Google Scholar 

  • Marshall, D.G., Bowe, F., Hale, C., Dougan, G., and Dorman, C.J. (2000). DNA topology and adaptation of Salmonella typhimurium to an intracellular environment. Phil. Trans. R. Soc. Lond. B. 355, 565–574.

    Article  CAS  Google Scholar 

  • Mekalanos, J.J. (1992). Environmental signals controlling expression of virulence determinants in bacteria. J. Bacteriol. 174, 1–7.

    CAS  PubMed  Google Scholar 

  • Merighi, M., Ellermeier, C.D., Slauch, J.M., and Gunn, J.S. (2005). Resolvase-in vivo expression technology analysis of the Salmo nella enterica serovar Typhimurium PhoP and PmrA regulons in BALB/c mice. J. Bacteriol. 187, 7407–7416.

    Article  CAS  PubMed  Google Scholar 

  • Monsieurs, P., De Keersmaecker, S., Navarre, W.W., Bader, M.W., De Smet, F., McClelland, M., Fang, F.C., De Moor, B., Vanderleyden, J., and Marchal, K. (2005). Comparison of the PhoPQ regulon in Escherichia coli and Salmonella typhimurium. J. Mol. Evol. 60, 462–474.

    Article  CAS  PubMed  Google Scholar 

  • Navarre, W.W., Halsey, T.A., Walthers, D., Frye, J., McClelland, M., Potter, J.L., Kenney, L.J., Gunn, J.S., Fang, F.C., and Libby, S.J. (2005). Co-regulation of Salmonella enterica genes required for virulence and resistance to antimicrobial peptides by SlyA and PhoP/PhoQ. Mol. Microbiol. 56, 492–508.

    Article  CAS  PubMed  Google Scholar 

  • Ohl, M.E., and Miller, S.I. (2001). Salmonella: A model for bacterial pathogenesis. Annu. Rev. Med. 52, 259–274.

    Article  CAS  PubMed  Google Scholar 

  • Pace, J., Hayman, M.J., and Galán, J.E. (1993). Signal transduction and invasion of epithelial cells by S. typhimurium. Cell 72, 505–514.

    Article  CAS  PubMed  Google Scholar 

  • Phillips, A.T., Egan, R.M., and Lewis, B. (1978). Control of biodegradative threonine dehydratase inducibility by cyclic AMP in energy-restricted Escherichia coli. J. Bacteriol. 35, 828–840.

    Google Scholar 

  • Sawers, G. (1998). The anaerobic degradation of l-serine and l-threonine in enterobacteria: networks of pathways and regulatory signals. Arch. Microbiol. 171, 1–5.

    Article  CAS  PubMed  Google Scholar 

  • Schmitt, C.K., Ikeda, J.S., Darnell, S.C., Watson, P.R., Bispham, J., Wallis, T.S., Weinstein, D.L., Metcalf, E.S., and O’Brien, A.D. (2001). Absence of all components of the flagellar export and synthesis machinery differentially alters virulence of Salmonella enterica serovar Typhimurium in models of typhoid fever, survival in macrophages, tissue culture invasiveness, and calf enterocolitis. Infect. Immun. 69, 5619–5625.

    Article  CAS  PubMed  Google Scholar 

  • Shin, D., Lee, E.-J., Huang, H., and Groisman, E.A. (2006). A positive feedback loop promotes transcription surge that jump-starts Salmonella virulence circuit. Science 314, 1607–1609.

    Article  CAS  PubMed  Google Scholar 

  • Shizuta, Y., and Hayaishi, O. (1970). Regulation of biodegradative threonine deaminase synthesis in Escherichia coli by cyclic adenosine-3′,5′-monophosphate. J. Biol. Chem. 245, 5416–5423.

    CAS  PubMed  Google Scholar 

  • Valdivia, R.H., and Falkow, S. (1997). Fluorescence-based isolation of bacterial genes expressed within host cells. Science 277, 2007–2011.

    Article  CAS  PubMed  Google Scholar 

  • Walthers, D., Carroll, R.K., Navarre, W.W., Libby, S.J., and Fang, F.C., and Kenney, L.J. (2007). The response regulator SsrB activates expression of diverse Salmonella pathogenecity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS. Mol. Microbiol. 65, 477–493.

    Article  CAS  PubMed  Google Scholar 

  • Yoon, H., Lim, S., Heu, S., Choi, S., and Ryu, S. (2003). Proteome of Salmonella enterica serovar Typhimurium fis mutant. FEMS Microbiol. Lett. 226, 391–396.

    Article  CAS  PubMed  Google Scholar 

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Affiliations

  1. Department of Food and Animal Biotechnology, Department of Agricultural Biotechnology, Center for Agricultural Biomaterials, and Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, Korea

    Minjeong Kim, Jeongjoon Choi & Sangryeol Ryu

  2. Radiation Research Division for Biotechnology, Korea Atomic Energy Research Institute, Jeongeup, 580-185, Korea

    Sangyong Lim

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  1. Sangyong Lim
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  2. Minjeong Kim
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  3. Jeongjoon Choi
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  4. Sangryeol Ryu
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Correspondence to Sangryeol Ryu.

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These authors contributed equally to this work.

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Lim, S., Kim, M., Choi, J. et al. A mutation in tdcA attenuates the virulence of Salmonella enterica serovar Typhimurium. Mol Cells 29, 509–517 (2010). https://doi.org/10.1007/s10059-010-0063-6

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  • DOI: https://doi.org/10.1007/s10059-010-0063-6

Keywords

  • Salmonella pathogenicity island 2 (SPI2)
  • ssaG
  • tdcA
  • virulence attenuation