CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression

Marat R. Sadykov, Ian H. Windham, Todd J. Widhelm, Vijaya Kumar Yajjala, Sean M. Watson, Jennifer L. Endres, Arissa I. Bavari, Vinai C. Thomas, Jeffrey L. Bose, Kenneth W. Bayles

Research output: Contribution to journalArticle

Abstract

The death and lysis of a subpopulation of Staphylococcus aureus cells during biofilm development benefit the whole bacterial population through the release of an important component of the biofilm matrix, extracellular DNA. Previously, we have demonstrated that these processes are affected by the gene products of the cidABC operon, the expression of which is controlled by the LysR-type transcriptional regulator, CidR. In this study, we characterized cis- and trans-acting elements essential for the induction of the cidABC operon. In addition to a CidR-binding site located within the cidABC promoter region, sequence analysis revealed the presence of a putative catabolite responsive element (cre box), suggestive of the involvement of the catabolite control protein A (CcpA) in the regulation of cidABC expression. This was confirmed using electrophoretic mobility shift assays and real-time reverse transcriptase PCR analysis demonstrating the direct positive control of cidABC transcription by the master regulator of carbon metabolism. Furthermore, the importance of CcpA and the identified cre site for the induction of the cidABC operon was demonstrated by examining the expression of PcidABC-lacZ reporter fusions in various mutant strains in which the genes involved in carbon metabolism and carbon catabolite repression were disrupted. Together the results of this study demonstrate the necessity of both transcriptional regulators, CidR and CcpA, for the induction of the cidABC operon and reveal the complexity of molecular interactions controlling its expression.

Original languageEnglish (US)
Article numbere00371-19
JournalJournal of bacteriology
Volume201
Issue number23
DOIs
StatePublished - Dec 1 2019

Fingerprint

Staphylococcal Protein A
Operon
Staphylococcus aureus
Biofilms
Carbon
Catabolite Repression
Electrophoretic Mobility Shift Assay
Reverse Transcriptase Polymerase Chain Reaction
Genetic Promoter Regions
Genes
Extracellular Matrix
Sequence Analysis
Real-Time Polymerase Chain Reaction
Binding Sites
DNA
Population

Keywords

  • Carbon catabolite repression
  • Glycolysis
  • Regulation of gene expression
  • Staphylococcus aureus

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Sadykov, M. R., Windham, I. H., Widhelm, T. J., Yajjala, V. K., Watson, S. M., Endres, J. L., ... Bayles, K. W. (2019). CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression. Journal of bacteriology, 201(23), [e00371-19]. https://doi.org/10.1128/JB.00371-19

CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression. / Sadykov, Marat R.; Windham, Ian H.; Widhelm, Todd J.; Yajjala, Vijaya Kumar; Watson, Sean M.; Endres, Jennifer L.; Bavari, Arissa I.; Thomas, Vinai C.; Bose, Jeffrey L.; Bayles, Kenneth W.

In: Journal of bacteriology, Vol. 201, No. 23, e00371-19, 01.12.2019.

Research output: Contribution to journalArticle

Sadykov, MR, Windham, IH, Widhelm, TJ, Yajjala, VK, Watson, SM, Endres, JL, Bavari, AI, Thomas, VC, Bose, JL & Bayles, KW 2019, 'CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression', Journal of bacteriology, vol. 201, no. 23, e00371-19. https://doi.org/10.1128/JB.00371-19
Sadykov MR, Windham IH, Widhelm TJ, Yajjala VK, Watson SM, Endres JL et al. CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression. Journal of bacteriology. 2019 Dec 1;201(23). e00371-19. https://doi.org/10.1128/JB.00371-19
Sadykov, Marat R. ; Windham, Ian H. ; Widhelm, Todd J. ; Yajjala, Vijaya Kumar ; Watson, Sean M. ; Endres, Jennifer L. ; Bavari, Arissa I. ; Thomas, Vinai C. ; Bose, Jeffrey L. ; Bayles, Kenneth W. / CidR and CcpA synergistically regulate staphylococcus aureus cidABC expression. In: Journal of bacteriology. 2019 ; Vol. 201, No. 23.
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