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Characterization of the sigmaE-dependent rpoEp3 promoter of Salmonella enterica serovar Typhimurium.
FEMS Microbiol Lett. 2006 Aug; 261(1):53-9.FM

Abstract

Using a two-plasmid system, we recently identified sigma(E)-dependent promoters directing expression of the sigma(E) regulon genes in Salmonella enterica serovar Typhimurium (S. Typhimurium). Comparison of the promoters revealed a consensus sequence almost identical to the sigma(E)-dependent rpoEp3 promoter directing expression of rpoE. This two-plasmid system was previously optimized to identify nucleotides critical for the rpoEp3 promoter activity. However, two highly conserved nucleotides in the sigma(E) consensus sequence were not identified by this screening. In the present study, we have improved the two-plasmid screening system using a new optimized error-prone PCR mutagenesis. Together with site-directed mutagenesis, we further identified nucleotides critical for activity of the rpoEp3 promoter and quantified the effect of the particular mutation upon promoter activity. All the identified critical nucleotides of the rpoEp3 promoter (in capital) were located in the -35 (ggAACtt) and -10 (gTCtaA) regions and corresponded to the most conserved nucleotides in the sigma(E) consensus sequence. The expression of the wild-type and mutated rpoEp3 promoters was confirmed in S. Typhimurium and was found to exhibit a different pattern of sigma(E) activation compared with Escherichia coli, with a peak rpoEp3 promoter activity in early stationary phase followed by a decrease in late stationary phase.

Authors+Show Affiliations

Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, Bratislava, Slovak Republik.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16842358

Citation

Skovierova, Henrieta, et al. "Characterization of the sigmaE-dependent rpoEp3 Promoter of Salmonella Enterica Serovar Typhimurium." FEMS Microbiology Letters, vol. 261, no. 1, 2006, pp. 53-9.
Skovierova H, Rezuchova B, Homerova D, et al. Characterization of the sigmaE-dependent rpoEp3 promoter of Salmonella enterica serovar Typhimurium. FEMS Microbiol Lett. 2006;261(1):53-9.
Skovierova, H., Rezuchova, B., Homerova, D., Roberts, M., & Kormanec, J. (2006). Characterization of the sigmaE-dependent rpoEp3 promoter of Salmonella enterica serovar Typhimurium. FEMS Microbiology Letters, 261(1), 53-9.
Skovierova H, et al. Characterization of the sigmaE-dependent rpoEp3 Promoter of Salmonella Enterica Serovar Typhimurium. FEMS Microbiol Lett. 2006;261(1):53-9. PubMed PMID: 16842358.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of the sigmaE-dependent rpoEp3 promoter of Salmonella enterica serovar Typhimurium. AU - Skovierova,Henrieta, AU - Rezuchova,Bronislava, AU - Homerova,Dagmar, AU - Roberts,Mark, AU - Kormanec,Jan, PY - 2006/7/18/pubmed PY - 2006/9/2/medline PY - 2006/7/18/entrez SP - 53 EP - 9 JF - FEMS microbiology letters JO - FEMS Microbiol Lett VL - 261 IS - 1 N2 - Using a two-plasmid system, we recently identified sigma(E)-dependent promoters directing expression of the sigma(E) regulon genes in Salmonella enterica serovar Typhimurium (S. Typhimurium). Comparison of the promoters revealed a consensus sequence almost identical to the sigma(E)-dependent rpoEp3 promoter directing expression of rpoE. This two-plasmid system was previously optimized to identify nucleotides critical for the rpoEp3 promoter activity. However, two highly conserved nucleotides in the sigma(E) consensus sequence were not identified by this screening. In the present study, we have improved the two-plasmid screening system using a new optimized error-prone PCR mutagenesis. Together with site-directed mutagenesis, we further identified nucleotides critical for activity of the rpoEp3 promoter and quantified the effect of the particular mutation upon promoter activity. All the identified critical nucleotides of the rpoEp3 promoter (in capital) were located in the -35 (ggAACtt) and -10 (gTCtaA) regions and corresponded to the most conserved nucleotides in the sigma(E) consensus sequence. The expression of the wild-type and mutated rpoEp3 promoters was confirmed in S. Typhimurium and was found to exhibit a different pattern of sigma(E) activation compared with Escherichia coli, with a peak rpoEp3 promoter activity in early stationary phase followed by a decrease in late stationary phase. SN - 0378-1097 UR - https://www.unboundmedicine.com/medline/citation/16842358/Characterization_of_the_sigmaE_dependent_rpoEp3_promoter_of_Salmonella_enterica_serovar_Typhimurium_ L2 - https://academic.oup.com/femsle/article-lookup/doi/10.1111/j.1574-6968.2006.00325.x DB - PRIME DP - Unbound Medicine ER -