In this study we characterized aro mutants of Salmonella enterica serovars Enteritidis and Typhimurium, which are frequently used as live oral vaccines. We found that the aroA, aroD, and aroC mutants were sensitive to blood serum, albumen, EDTA, and ovotransferrin, and this defect could be complemented by an appropriate aro gene cloned in a plasmid. Subsequent microarray analysis of gene expression in the aroD mutant in serovar Typhimurium indicated that the reason for this sensitivity might be the upregulation of murA. To confirm this, we artificially overexpressed murA from a multicopy plasmid, and this overexpression caused sensitivity of the strain to albumen and EDTA but not to serum and ovotransferrin. We concluded that attenuation of aro mutants is caused not only by their inability to synthesize aromatic metabolites but also by their defect in cell wall and outer membrane functions associated with decreased resistance to components of innate immune response.

Veterinary Research Institute Hudcova 70 621 00 Brno Czech Republic

Zobrazit více v PubMed

Bar-Tana, J., B. J. Howlett, and R. Hertz. 1980. Ubiquinone synthetic pathway in flagellation of PubMed PMC

Bennett, G. M., A. Seaver, and P. H. Calcott. 1981. Effect of defined lipopolysaccharide core defects on resistance of PubMed PMC

Brown, E. D., E. I. Vivas, C. T. Walsh, and R. Kolter. 1995. MurA (MurZ), the enzyme that catalyzes the first committed step in peptidoglycan biosynthesis, is essential in PubMed PMC

Conlan, J. W., and R. J. North. 1992. Early pathogenesis of infection in the liver with the facultative intracellular bacteria PubMed PMC

Cooper, G. L., L. M. Venables, M. J. Woodward, and C. E. Hormaeche. 1994. Vaccination of chickens with strain CVL30, a genetically defined PubMed PMC

Datsenko, K. A., and B. L. Wanner. 2000. One-step inactivation of chromosomal genes in PubMed PMC

Duncan, K., S. Chaudhuri, M. S. Campbell, and J. R. Coggins. 1986. The overexpression and complete amino acid sequence of PubMed PMC

Duncan, K., and J. R. Coggins. 1986. The PubMed PMC

Eschenburg, S., W. Kabsch, M. L. Healy, and E. Schonbrunn. 2003. A new view of the mechanisms of UDP- PubMed

Everest, P., J. Ketley, S. Hardy, G. Douce, S. Khan, J. Shea, D. Holden, D. Maskell, and G. Dougan. 1999. Evaluation of PubMed PMC

Galloway, S. M., and C. R. Raetz. 1990. A mutant of PubMed

Govoni, G., and P. Gros. 1998. Macrophage NRAMP1 and its role in resistance to microbial infections. Inflamm. Res. 47277-284. PubMed

Halsey, T. A., A. Vazquez-Torres, D. J. Gravdahl, F. C. Fang, and S. J. Libby. 2004. The ferritin-like Dps protein is required for PubMed PMC

Hoiseth, S. K., and B. A. D. Stocker. 1981. Aromatic-dependent PubMed

Hoiseth, S. K., and B. A. Stocker. 1985. Genes PubMed PMC

Horii, T., T. Kimura, K. Sato, K. Shibayama, and M. Ohta. 1999. Emergence of fosfomycin-resistant isolates of Shiga-like toxin-producing PubMed PMC

Hormaeche, C. E., H. S. Joysey, L. Desilva, M. Izhar, and B. A. Stocker. 1991. Immunity conferred by Aro PubMed

Lowe, D. C., T. C. Savidge, D. Pickard, L. Eckmann, M. F. Kagnoff, G. Dougan, and S. N. Chatfield. 1999. Characterization of candidate live oral PubMed PMC

Methner, U., P. A. Barrow, D. Gregorova, and I. Rychlik. 2004. Intestinal colonization-inhibition and virulence of PubMed

Nnalue, N. A. 1990. Mice vaccinated with a non-virulent, aromatic-dependent mutant of PubMed

Perez-Gutierrez, C., C. M. Llompart, M. Skurnik, and J. A. Bengoechea. 2007. Expression of the PubMed PMC

Robertsson, J. A., A. A. Lindberg, S. Hoiseth, and B. A. Stocker. 1983. PubMed PMC

Rychlik, I., L. Cardova, M. Sevcik, and P. A. Barrow. 2000. Flow cytometry characterisation of PubMed

Rychlik, I., G. Martin, U. Methner, M. Lovell, L. Cardova, A. Sebkova, M. Sevcik, J. Damborsky, and P. A. Barrow. 2002. Identification of PubMed

Shaio, M. F., and H. Rowland. 1985. Bactericidal and opsonizing effects of normal serum on mutant strains of PubMed PMC

Silva, E. N., G. H. Snoeyenbos, O. M. Weinack, and C. F. Smyser. 1981. Studies on the use of 9R strain of PubMed

Splichal, I., I. Trebichavsky, A. Splichalova, and P. A. Barrow. 2005. Protection of gnotobiotic pigs against PubMed

Stevenson, G., B. Neal, D. Liu, M. Hobbs, N. H. Packer, M. Batley, J. W. Redmond, L. Lindquist, and P. Reeves. 1994. Structure of the O antigen of PubMed PMC

Stinavage, P., L. E. Martin, and J. K. Spitznagel. 1989. O antigen and lipid A phosphoryl groups in resistance of PubMed PMC

Stritzker, J., J. Janda, C. Schoen, M. Taupp, S. Pilgrim, I. Gentschev, P. Schreier, G. Geginat, and W. Goebel. 2004. Growth, virulence, and immunogenicity of PubMed PMC

Tacket, C. O., M. B. Sztein, G. A. Losonsky, S. S. Wasserman, J. P. Nataro, R. Edelman, D. Pickard, G. Dougan, S. N. Chatfield, and M. M. Levine. 1997. Safety of live oral PubMed PMC

Tacket, C. O., M. B. Sztein, S. S. Wasserman, G. Losonsky, K. L. Kotloff, T. L. Wyant, J. P. Nataro, R. Edelman, J. Perry, P. Bedford, D. Brown, S. Chatfield, G. Dougan, and M. M. Levine. 2000. Phase 2 clinical trial of attenuated PubMed PMC

Trebichavsky, I., A. Splichalova, I. Rychlik, H. Hojna, Y. Muneta, Y. Mori, and I. Splichal. 2006. Attenuated PubMed

Tusher, V. G., R. Tibshirani, and G. Chu. 2001. Significance analysis of microarrays applied to the ionizing radiation response. Proc. Natl. Acad. Sci. USA 985116-5121. PubMed PMC

phoP, SPI1, SPI2 and aroA mutants of Salmonella Enteritidis induce a different immune response in chickens