The sigma factor RpoN (σ54) is involved in osmotolerance in Listeria monocytogenes
Yumiko Okada
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorNobuhiko Okada
Department of Microbiology, Kitasato University School of Pharmaceutical Sciences, Tokyo 108-8641, Japan
Search for more papers by this authorSou-ichi Makino
Laboratory of Food Microbiology and Immunology, Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan
Search for more papers by this authorHiroshi Asakura
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorShigeki Yamamoto
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorShizunobu Igimi
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorYumiko Okada
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorNobuhiko Okada
Department of Microbiology, Kitasato University School of Pharmaceutical Sciences, Tokyo 108-8641, Japan
Search for more papers by this authorSou-ichi Makino
Laboratory of Food Microbiology and Immunology, Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido 080-8555, Japan
Search for more papers by this authorHiroshi Asakura
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorShigeki Yamamoto
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorShizunobu Igimi
Division of Biomedical Food Research, National Instutute of Health Sciences, Tokyo 158–8501, Japan
Search for more papers by this authorEditor: Aharon Oren
Abstract
Listeria monocytogenes is able to grow under conditions of high osmolarity. We constructed a deletion mutant of rpoN, encoding the alternative sigma factor RpoN, and analyzed its response to osmotic stress. In a minimal medium with 4% NaCl and 1 mM betaine, the mutant showed a similar growth to that of the parental strain, EGD. In the same medium with 4% NaCl and 1 M carnitine, the growth rate of the mutant was greatly reduced, when the optical density at 600 nm (OD600) at the starting point of growth, was 0.15. However, when growth of the culture was started at an OD600 of 0.025, the growth of the mutant was similar to that of EGD. The mutant's expression of two betaine transporter genes, betL and gbuB, and the carnitine transporter gene opuCA, was osmotically induced at a level similar to EGD, and its rate of carnitine uptake was similar to that of EGD. These results suggest that the growth defect from the rpoN mutant is caused not by the transcriptional regulation of opuCA or by a decrease in carnitine uptake, but possibly by larger amounts of carnitine being needed for growth of the mutant in minimal medium when NaCl is present.
References
- Ali NO, Bignon J, Rapoport G & Debarbouille M (2001) Regulation of the acetoin catabolic pathway is controlled by sigma L in Bacillus subtilis. J Bacteriol 183: 2497–2504.
- Arous S, Buchrieser C, Folio P, Glaser P, Namane A, Hebraud M & Hechard Y (2004) Global analysis of gene expression in an rpoN mutant of Listeria monocytogenes. Microbiol 150: 1581–1590.
- Becker LA, Cetin MH, Hutkins RW & Benson AK (1998) Identification of the gene encoding the alternative sigma factor B from Listeria monocytogenes and its role in osmotolerance. J Bacteriol 180: 4547–4554.
- Becker LA, Evans SN, Hutkins RW & Benson AK (2000) Role of sigma (B) in adaptation of Listeria monocytogenes to growth at low temperature. J Bacteriol 182: 7083–7087.
- Beumer RR, Te Giffel MC, Cox LJ, Rombouts FM & Abee T (1994) Effect of exogenous proline, betaine, and carnitine on growth of Listeria monocytogenes in a minimal medium. Appl Environ Microbiol 60: 1359–1363.
- Cetin MS, Zhang C, Hutkins RW & Benson AK (2004) Regulation of transcription of compatible solute transporters by the general stress sigma factor, σB, in Listeria monocytogenes. J Bacteriol 186: 794–802.
- Dalet K, Cenatiempo Y, Cossart P & Hechard Y (2001) The European Listeria Genome ConsortiumA σ54-dependent PTS permease of the mannose family is responsible for sensitivity of Listeria monocytogenes to mesentericin Y105. Microbiol 147: 3263–3269.
- Dalet K, Arous S, Cenatiempo Y & Hechard Y (2003) Characterization of a unique σ54-dependent PTS operon of the lactose family in Listeria monocytogenes. Biochimie 85: 633–638.
- Debarbouille M, Martin-Verstraete I, Kunst F & Rapoport G (1991) The Bacillus subtilis sigL gene encodes an equivalent of σ54 from Gram-negative bacteria. Proc Natl Acad Sci USA 88: 9092–9296.
- Fraser KR, Harvie D, Coote PJ & O'Byrne CP (2000) Identification and characterization of an ATP binding cassette l-carnitine transporter in Listeria monocytogenes. Appl Environ Microbiol 66: 4696–4704.
- Fraser KR, Sue D, Wiedmann M, Boor K & O'Byrne CP (2003) Role of σB in regulating the compatible solute uptake systems of Listeria monocytogenes: osmotic induction of opuC is σB dependent. Appl Environ Microbiol 69: 2015–2022.
- Glaser P, Frangeul L, Buchreiser C et al. (2001) Comparative genomics of Listeria species. Science 294: 849–852.
- Ishiwa H & Shibahara H (1985) New shuttle vectors for Escherichia coli and Bacillus subtilis. III. Nucleotide sequence analysis of tetracycline resistance gene of pAMalpha1 and ori-177. Jpn J Genet 60: 485–498.
- Jishage M, Iwata A, Ueda S & Ishihama A (1996) Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions. J Bacteriol 178: 5447–5451.
- Ko R & Smith LT (1999) Identification of an ATP-driven, osmoregulated glycine betaine transport system in Listeria monocytogenes. Appl Environ Microbiol 65: 4040–4048.
- Kohler T, Harayama S, Ramos JL & Timmis KN (1989) Involvement of Pseudomonas putida RpoN sigma factor in regulation of various metabolic functions. J Bacteriol 171: 4326–4333.
- Metzger R, Brown DP, Grealish P, Staver MJ, Versalovic J, Lupski JR & Katz L (1994) Characterization of the macromolecular synthesis (MMS) operon from Listeria monocytogenes. Gene 151: 161–166.
- Okada Y, Makino S, Tobe T, Okada N & Yamazaki S (2002) Cloning of rel from Listeria monocytogenes as an osmotolerance involvement gene. Appl Environ Microbiol 68: 1541–1547.
- Okutani A, Okada Y, Yamamoto S & Igimi S (2004) Overview of Listeria monocytogenes contamination in Japan. Int J Food Microbiol 92: 131–140.
- Park SF & Stewart GS (1990) High-efficiency transformation of Listeria monocytogenes by electroporation of penicillin-treated cells. Gene 94: 129–132.
-
Phan-Thanh L &
Mahouin F (1999) A proteomic approach to study the acid response in Listeria monocytogenes.
Electrophoresis
20: 2214–2224.
10.1002/(SICI)1522-2683(19990801)20:11<2214::AID-ELPS2214>3.0.CO;2-G CAS PubMed Web of Science® Google Scholar
- Rea RB, Gahan CGM & Hill C (2004) Disruption of putative regulatory loci in Listeria monocytogenes demonstrates a significant role for Fur and PerR in virulence. Infect Immun 72: 717–727.
- Robichon D, Gouin E, Debarbouille M, Cossart P, Cenatiempo Y & Hechard Y (1997) The rpoN (sigma54) gene from Listeria monocytogenes is involved in resistance to mesentericin Y105, an antibacterial peptide from Leuconostoc mesenteroides. J Bacteriol 179: 7591–7594.
-
Sambrook J,
Fritsch EF &
Maniatis T. (1989) Molecular Cloning: A Laboratory Manual, 2nd edn.
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
10.1111/j.1095-8312.1996.tb01434.x Google Scholar
- Seeliger HPR & Jones D (1986) Listeria. Bergey's Manual of Systematic Bacteriology, Vol. 2. ( PHA Sneath, NS Mair, ME Sharpe & JG Holt, eds), pp. 1235–1245. Williams & Wilkins, Baltimore, MD.
- Sleator RD, Gahan CG, Abee T & Hill C (1999) Identification and disruption of BetL, a secondary glycine betaine transport system linked to the salt tolerance of Listeria monocytogenes LO28. Appl Environ Microbiol 65: 2078–2083.
- Sleator RD, Gahan CG & Hill C (2003a) A postgenomic appraisal of osmotolerance in Listeria monocytogenes. Appl Environ Microbiol 69: 1–9.
- Sleator RD, Wood JM & Hill C (2003b) Transcriptional regulation and posttranslational activity of the betaine transporter BetL in Listeria monocytogenes are controlled by environmental salinity. J Bacteriol 185: 7140–7144.
- Sullivan MA, Yasbin RE & Young FE (1984) New shuttle vectors for Bacillus subtilis and Escherichia coli which allow rapid detection of inserted fragments. Gene 29: 21–26.
- Vadyvaloo V, Arous S, Gravesen A, Hechard Y, Chauhan-Haubrock R, Hastings JW & Rautenbach M (2004) Cell-surface alterations in class IIa bacteriocin-resistant Listeria monocytogenes strains. Microbiology 150: 3025–3033.
- Verheul A, Glaasker E, Poolman B & Abee T (1997) Betaine and l-carnitine transport by Listeria monocytogenes Scott A in response to osmotic signals. J Bacteriol 179: 6979–6985.
- Walker SJ, Archer P & Banks JG (1990) Growth of Listeria monocytogenes at refrigeration temperatures. J Appl Bacteriol 68: 157–162.
- Wiedmann M, Arvik TJ, Hurley RJ & Boor KJ (1998) General stress transcription factor B and its role in acid tolerance and virulence of Listeria monocytogenes. J Bacteriol 180: 3650–3656.
- Youngman P (1987) Plasmid vectors for recovering and exploiting Tn917 transpositions in Bacillus and other Gram-positive bacteria. Plasmids, A Practical Approach ( KG Hardy, ed), pp. 79–104. IRL Press Ltd., Oxford.