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Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Staphylococcus aureus alternative sigma factor σB

Homerova, Dagmar ; Bischoff, Markus ; Dumolin, Alexis ; Kormanec, Jan

In: FEMS Microbiology Letters, 2004, vol. 232, no. 2, p. 173-179

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    Titre
    • Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Staphylococcus aureus alternative sigma factor σB
    Auteur
    • Homerova, Dagmar. Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovak Republic
    • Bischoff, Markus. Institute of Medical Microbiology, University of Zürich, CH-8028 Zürich, Switzerland
    • Dumolin, Alexis. Institute of Medical Microbiology, University of Zürich, CH-8028 Zürich, Switzerland
    • Kormanec, Jan. Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovak Republic
    Type de document
    • Postprint
    Identifiant OAI-PMH
    • oai:doc.rero.ch:292719
    Summary
    We optimized a previously established two-plasmid system for the identification of Staphylococcus aureus promoters that are recognized by the alternative transcription factor σB. The method allowed the identification of 18 S. aureusσB-dependent promoters, 12 of which are reported here for the first time to be σB-dependent. S1-nuclease mapping of the respective transcriptional start points revealed that all the promoters contained sequences exhibiting high similarity to the consensus sequence of Bacillus subtilisσB-dependent promoters. The promoters governed expression of genes encoding proteins proposed to be involved in various cellular functions, including the stress response genes and virulence-associated clfA gene for fibrinogen-binding clumping factor. Comparison of the nucleotide sequences upstream of the identified transcription start points identified a σB consensus promoter (GttTaa-N12-15-gGGTAt) that is highly homologous to that of σB of B. subtilis