Abstract
The osmotically regulated proU locus in Escherichia coli has two promoters, P1 and P2, that are recognized, respectively, by the sigmaS- and sigma70-bearing RNA polymerase holoenzymes. However, the equivalent of the P1 promoter does not appear to exist in Salmonella typhimurium. We demonstrate in this study that wild-type S. typhimurium has a cryptic P1 promoter that is recognized by sigmaS RNA polymerase in vitro and that a 22-bp deletion from +63 to +84 (relative to the start site of transcription) confers sigmaS-dependent in vivo expression of a reporter gene fusion to P1. Primer extension analysis of RNA isolated from cells carrying the wild-type and mutant S. typhimurium proU constructs indicated that a primer which hybridizes proximal to +60 is able to detect P1-initiated transcripts from both constructs but a primer which hybridizes distal to +85 is able to do so only from the latter. Our results suggest that the sigmaS-controlled proU P1 promoter in S. typhimurium may be rendered cryptic because of factor-dependent transcription attenuation within a short distance downstream of the promoter start site.
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- Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
- Andrews A. E., Lawley B., Pittard A. J. Mutational analysis of repression and activation of the tyrP gene in Escherichia coli. J Bacteriol. 1991 Aug;173(16):5068–5078. doi: 10.1128/jb.173.16.5068-5078.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Conway T., Osman Y. A., Konnan J. I., Hoffmann E. M., Ingram L. O. Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase. J Bacteriol. 1987 Mar;169(3):949–954. doi: 10.1128/jb.169.3.949-954.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dattananda C. S., Rajkumari K., Gowrishankar J. Multiple mechanisms contribute to osmotic inducibility of proU operon expression in Escherichia coli: demonstration of two osmoresponsive promoters and of a negative regulatory element within the first structural gene. J Bacteriol. 1991 Dec;173(23):7481–7490. doi: 10.1128/jb.173.23.7481-7490.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gollnick P. Regulation of the Bacillus subtilis trp operon by an RNA-binding protein. Mol Microbiol. 1994 Mar;11(6):991–997. doi: 10.1111/j.1365-2958.1994.tb00377.x. [DOI] [PubMed] [Google Scholar]
- Gowrishankar J., Jayashree P., Rajkumari K. Molecular cloning of an osmoregulatory locus in Escherichia coli: increased proU gene dosage results in enhanced osmotolerance. J Bacteriol. 1986 Dec;168(3):1197–1204. doi: 10.1128/jb.168.3.1197-1204.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gowrishankar J., Manna D. How is osmotic regulation of transcription of the Escherichia coli proU operon achieved? A review and a model. Genetica. 1996 May;97(3):363–378. doi: 10.1007/BF00055322. [DOI] [PubMed] [Google Scholar]
- Gowrishankar J. Nucleotide sequence of the osmoregulatory proU operon of Escherichia coli. J Bacteriol. 1989 Apr;171(4):1923–1931. doi: 10.1128/jb.171.4.1923-1931.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kolb A., Kotlarz D., Kusano S., Ishihama A. Selectivity of the Escherichia coli RNA polymerase E sigma 38 for overlapping promoters and ability to support CRP activation. Nucleic Acids Res. 1995 Mar 11;23(5):819–826. doi: 10.1093/nar/23.5.819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kundu T. K., Kusano S., Ishihama A. Promoter selectivity of Escherichia coli RNA polymerase sigmaF holoenzyme involved in transcription of flagellar and chemotaxis genes. J Bacteriol. 1997 Jul;179(13):4264–4269. doi: 10.1128/jb.179.13.4264-4269.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu M. Y., Yang H., Romeo T. The product of the pleiotropic Escherichia coli gene csrA modulates glycogen biosynthesis via effects on mRNA stability. J Bacteriol. 1995 May;177(10):2663–2672. doi: 10.1128/jb.177.10.2663-2672.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Loewen P. C., Hengge-Aronis R. The role of the sigma factor sigma S (KatF) in bacterial global regulation. Annu Rev Microbiol. 1994;48:53–80. doi: 10.1146/annurev.mi.48.100194.000413. [DOI] [PubMed] [Google Scholar]
- Lu Y., Switzer R. L. Evidence that the Bacillus subtilis pyrimidine regulatory protein PyrR acts by binding to pyr mRNA at three sites in vivo. J Bacteriol. 1996 Oct;178(19):5806–5809. doi: 10.1128/jb.178.19.5806-5809.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lucht J. M., Bremer E. Adaptation of Escherichia coli to high osmolarity environments: osmoregulation of the high-affinity glycine betaine transport system proU. FEMS Microbiol Rev. 1994 May;14(1):3–20. doi: 10.1111/j.1574-6976.1994.tb00067.x. [DOI] [PubMed] [Google Scholar]
- Manna D., Gowrishankar J. Evidence for involvement of proteins HU and RpoS in transcription of the osmoresponsive proU operon in Escherichia coli. J Bacteriol. 1994 Sep;176(17):5378–5384. doi: 10.1128/jb.176.17.5378-5384.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mellies J., Brems R., Villarejo M. The Escherichia coli proU promoter element and its contribution to osmotically signaled transcription activation. J Bacteriol. 1994 Jun;176(12):3638–3645. doi: 10.1128/jb.176.12.3638-3645.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Overdier D. G., Csonka L. N. A transcriptional silencer downstream of the promoter in the osmotically controlled proU operon of Salmonella typhimurium. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):3140–3144. doi: 10.1073/pnas.89.7.3140. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Overdier D. G., Olson E. R., Erickson B. D., Ederer M. M., Csonka L. N. Nucleotide sequence of the transcriptional control region of the osmotically regulated proU operon of Salmonella typhimurium and identification of the 5' endpoint of the proU mRNA. J Bacteriol. 1989 Sep;171(9):4694–4706. doi: 10.1128/jb.171.9.4694-4706.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rajkumari K., Kusano S., Ishihama A., Mizuno T., Gowrishankar J. Effects of H-NS and potassium glutamate on sigmaS- and sigma70-directed transcription in vitro from osmotically regulated P1 and P2 promoters of proU in Escherichia coli. J Bacteriol. 1996 Jul;178(14):4176–4181. doi: 10.1128/jb.178.14.4176-4181.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stirling D. A., Hulton C. S., Waddell L., Park S. F., Stewart G. S., Booth I. R., Higgins C. F. Molecular characterization of the proU loci of Salmonella typhimurium and Escherichia coli encoding osmoregulated glycine betaine transport systems. Mol Microbiol. 1989 Aug;3(8):1025–1038. doi: 10.1111/j.1365-2958.1989.tb00253.x. [DOI] [PubMed] [Google Scholar]
- Tanaka K., Kusano S., Fujita N., Ishihama A., Takahashi H. Promoter determinants for Escherichia coli RNA polymerase holoenzyme containing sigma 38 (the rpoS gene product). Nucleic Acids Res. 1995 Mar 11;23(5):827–834. doi: 10.1093/nar/23.5.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tomizawa J., Itoh T., Selzer G., Som T. Inhibition of ColE1 RNA primer formation by a plasmid-specified small RNA. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1421–1425. doi: 10.1073/pnas.78.3.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ueguchi C., Mizuno T. The Escherichia coli nucleoid protein H-NS functions directly as a transcriptional repressor. EMBO J. 1993 Mar;12(3):1039–1046. doi: 10.1002/j.1460-2075.1993.tb05745.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wise A., Brems R., Ramakrishnan V., Villarejo M. Sequences in the -35 region of Escherichia coli rpoS-dependent genes promote transcription by E sigma S. J Bacteriol. 1996 May;178(10):2785–2793. doi: 10.1128/jb.178.10.2785-2793.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]
- Yanofsky C., Konan K. V., Sarsero J. P. Some novel transcription attenuation mechanisms used by bacteria. Biochimie. 1996;78(11-12):1017–1024. doi: 10.1016/s0300-9084(97)86725-9. [DOI] [PubMed] [Google Scholar]
- Yim H. H., Brems R. L., Villarejo M. Molecular characterization of the promoter of osmY, an rpoS-dependent gene. J Bacteriol. 1994 Jan;176(1):100–107. doi: 10.1128/jb.176.1.100-107.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang X., Fletcher S. A., Csonka L. N. Site-directed mutational analysis of the osmotically regulated proU promoter of Salmonella typhimurium. J Bacteriol. 1996 Jun;178(11):3377–3379. doi: 10.1128/jb.178.11.3377-3379.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]