Abstract
Salmonella enterica serovar Typhi z66 positive strain contains a fljBA-like operon on a linear plasmid. The operon contains the gene fljB:z66 which encodes the z66 antigen. RpoE is a sigma factor σE that initiates transcription of a series of genes in Escherichia and Salmonella under environmental stresses. To investigate whether the gene fljB:z66 is regulated by RpoE (σE), a rpoE deletion mutant of S. enterica serovar Typhi (ΔrpoE) was prepared in this study. The defective motility of the ΔrpoE was confirmed firstly. Transcriptional expression of flagellar genes was screened using a genomic DNA microarray. Some class-2 and most class-3 flagellar genes were downregulated in the ΔrpoE after 30 min of hyperosmotic stress. The expression of fliA and fljB:z66, a class-2 flagellar gene and a class-3 flagellar gene, obviously decreased; however, expression of the class-1 flagellar genes flhDC did not change obviously in the ΔrpoE compared to the wild-type strain in the same conditions. Results of quantitative real-time PCR (qRT-PCR) showed that the expression levels of fliA and fljB:z66 in the ΔrpoE after 30 min of hyperosmotic stress decreased about five and eightfold, respectively, compared to the wild-type strain. Similar results were observed at 120 min of hyperosmotic stress. Western blotting and qRT-PCR analysis showed that expression of fliA and fljB:z66 was significantly increased after supplemental expression of rpoE with a recombinant plasmid pBADrpoE in the ΔrpoE strain. These results demonstrated that RpoE promoted the expression of class-3 flagellar genes and it might be performed by initiating the expression of fliA in S. enterica serovar Typhi under hyperosmotic stress.
Similar content being viewed by others
References
Alba BM, Gross CA (2004) Regulation of the Escherichia coli sigma-dependent envelope stress response. Mol Microbiol 52:613–619
Alba BM, Leeds JA, Onufryk C et al (2002) DegS and YaeL participate sequentially in the cleavage of RseA to activate the sigma(E)-dependent extracytoplasmic stress response. Genes Dev 16:2156–2168
Alba BM, Zhong HJ, Pelayo JC, Gross CA (2001) degS (hhoB) is an essential Escherichia coli gene whose indispensable function is to provide sigma (E) activity. Mol Microbiol 40:1323–1333
Baker S, Hardy J, Sanderson KE et al (2007) A novel linear plasmid mediates flagellar variation in Salmonella Typhi. PLoS Pathog 3:e59
Baker S, Holt K, Whitehead S et al (2007) A linear plasmid truncation induces unidirectional flagellar phase change in H:z66 positive Salmonella Typhi. Mol Microbiol 66:1207–1218
Everest P, Wain J, Roberts M et al (2001) The molecular mechanisms of severe typhoid fever. Trends Microbiol 9:316–320
Frankel G, Newton SM, Schoolnik GK, Stocker BA (1989) Intragenic recombination in a flagellin gene: characterization of the H1-j gene of Salmonella typhi. EMBO J 8:3149–3152
Grigorova IL, Chaba R, Zhong HJ et al (2004) Fine-tuning of the Escherichia coli sigmaE envelope stress response relies on multiple mechanisms to inhibit signal-independent proteolysis of the transmembrane anti-sigma factor, RseA. Genes Dev 18:2686–2697
Hayden JD, Ades SE (2008) The extracytoplasmic stress factor, sigmaE, is required to maintain cell envelope integrity in Escherichia coli. PLoS ONE 3:e1573
Huang X, Phung le V, Dejsirilert S et al (2004) Cloning and characterization of the gene encoding the z66 antigen of Salmonella enterica serovar Typhi. FEMS Microbiol Lett 234:239–246
Huang X, Xu H, Sun X et al (2007) Genome-wide scan of the gene expression kinetics of Salmonella enterica serovar Typhi during hyperosmotic stress. Int J Mol Sci 8:116–135
Jones BD, Lee CA, Falkow S (1992) Invasion by Salmonella typhimurium is affected by the direction of flagellar rotation. Infect Immun 60:2475–2480
Kutsukake K, Iino T (1994) Role of the FliA-FlgM regulatory system on the transcriptional control of the flagellar regulon and flagellar formation in Salmonella typhimurium. J Bacteriol 176:3598–3605
Kutsukake K, Ohya Y, Iino T (1990) Transcriptional analysis of the flagellar regulon of Salmonella typhimurium. J Bacteriol 172:741–747
Leclerc GJ, Tartera C, Metcalf ES (1998) Environmental regulation of Salmonella typhi invasion-defective mutants. Infect Immun 66:682–691
Liu SL, Ezaki T, Miura H et al (1988) Intact motility as a Salmonella typhi invasion-related factor. Infect Immun 56:1967–1973
Mashimo T, Hashimoto M, Yamaguchi S, Aizawa S (2007) Temperature-hypersensitive sites of the flagellar switch component FliG in Salmonella enterica serovar typhimurium. J Bacteriol 189:5153–5160
Miticka H, Rowley G, Rezuchova B et al (2003) Transcriptional analysis of the rpoE gene encoding extracytoplasmic stress response sigma factor sigmaE in Salmonella enterica serovar Typhimurium. FEMS Microbiol Lett 226:307–314
Pickard D, Li J, Roberts M et al (1994) Characterization of defined ompR mutants of Salmonella typhi: ompR is involved in the regulation of Vi polysaccharide expression. Infect Immun 62:3984–3993
Rhodius VA, Suh WC, Nonaka G et al (2006) Conserved and variable functions of the sigmaE stress response in related genomes. PLoS Biol 4:e2
Rolhion N, Carvalho FA, Darfeuille-Michaud A (2007) OmpC and the sigma(E) regulatory pathway are involved in adhesion and invasion of the Crohn’s disease-associated Escherichia coli strain LF82. Mol Microbiol 63:1684–1700
Sheng X, Huang X, Mao L et al (2009) Preparation of Salmonella enterica serovar Typhi genomic DNA microarrays for gene expression profiling analysis. Prog Biochem Biophys 36:306–312
Shin S, Park C (1995) Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR. J Bacteriol 177:4696–4702
Tartera C, Metcalf ES (1993) Osmolarity and growth phase overlap in regulation of Salmonella typhi adherence to and invasion of human intestinal cells. Infect Immun 61:3084–3089
Testerman TL, Vazquez-Torres A, Xu Y et al (2002) The alternative sigma factor sigmaE controls antioxidant defences required for Salmonella virulence and stationary-phase survival. Mol Microbiol 43:771–782
Tusher V, Tibshirani R, Chu C (2001) Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA 98:5116–5512
Wang Q, Suzuki A, Mariconda S et al (2005) Sensing wetness: a new role for the bacterial flagellum. EMBO J 24:2034–2042
Xu S, Zou X, Sheng X et al (2010) Expression of fljB:z66 on a linear plasmid of Salmonella enterica serovar Typhi is dependent on FliA and FlhDC and regulated by OmpR. Braz J Microbiol (accepted)
Zhang H, Sheng X, Xu S et al (2009) Global transcriptional response of Salmonella enterica serovar Typhi to anti-z66 antiserum. FEMS Microbiol Lett 298:51–55
Zou X, Huang X, Xu S et al (2009) Identification of a fljA gene on a linear plasmid as the repressor gene of fliC in Salmonella enterica serovar Typhi. Microbiol Immunol 53:191–197
Acknowledgments
We thank T. Ezaki (Gifu University) for bacterial strains, anti-z66 antiserum, and continuous support. This study was supported by National Natural Science Foundation of China (30870095), National Special Scientific Program (2008ZX10004-009), Science Foundation of Jiangsu University for Advanced Scholars and Special Team Program of Jiangsu University (2008-018-02).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Du, H., Sheng, X., Zhang, H. et al. RpoE may Promote Flagellar Gene Expression in Salmonella enterica Serovar Typhi Under Hyperosmotic Stress. Curr Microbiol 62, 492–500 (2011). https://doi.org/10.1007/s00284-010-9734-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00284-010-9734-y