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Characterization of the SarA virulence gene regulator of Staphylococcus aureus.

Publication ,  Journal Article
Rechtin, TM; Gillaspy, AF; Schumacher, MA; Brennan, RG; Smeltzer, MS; Hurlburt, BK
Published in: Mol Microbiol
July 1999

Staphylococcus aureus is a potent human pathogen that expresses a large number of virulence factors in a temporally regulated fashion. Two pleiotropically acting regulatory loci were identified in previous mutational studies. The agr locus comprises two operons that express a quorum-sensing system from the P2 promoter and a regulatory RNA molecule from the P3 promoter. The sar locus encodes a DNA-binding protein that activates the expression of both agr operons. We have cloned the sarA gene, expressed SarA in Escherichia coli and purified the recombinant protein to apparent homogeneity. The purified protein was found to be dimeric in the presence and absence of DNA and to consist mostly of alpha-helices. DNase I footprinting of SarA on the putative regulatory region cis to the agr promoters revealed three high-affinity binding sites composed of two half-sites each. Quantitative electrophoretic mobility shift assays (EMSAs) were used to derive equilibrium binding constants (KD) for the interaction of SarA with these binding sites. An unusual ladder banding pattern was observed in EMSA with a large DNA fragment including all three binding sites. Our data indicate that SarA regulation of the agr operons involves binding to multiple half-sites and may involve other sites located downstream of the promoters.

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Published In

Mol Microbiol

DOI

ISSN

0950-382X

Publication Date

July 1999

Volume

33

Issue

2

Start / End Page

307 / 316

Location

England

Related Subject Headings

  • Trans-Activators
  • Staphylococcus aureus
  • Protein Conformation
  • Molecular Sequence Data
  • Microbiology
  • Gene Expression Regulation, Bacterial
  • Dimerization
  • DNA Footprinting
  • Circular Dichroism
  • Base Sequence
 

Citation

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Rechtin, T. M., Gillaspy, A. F., Schumacher, M. A., Brennan, R. G., Smeltzer, M. S., & Hurlburt, B. K. (1999). Characterization of the SarA virulence gene regulator of Staphylococcus aureus. Mol Microbiol, 33(2), 307–316. https://doi.org/10.1046/j.1365-2958.1999.01474.x
Rechtin, T. M., A. F. Gillaspy, M. A. Schumacher, R. G. Brennan, M. S. Smeltzer, and B. K. Hurlburt. “Characterization of the SarA virulence gene regulator of Staphylococcus aureus.Mol Microbiol 33, no. 2 (July 1999): 307–16. https://doi.org/10.1046/j.1365-2958.1999.01474.x.
Rechtin TM, Gillaspy AF, Schumacher MA, Brennan RG, Smeltzer MS, Hurlburt BK. Characterization of the SarA virulence gene regulator of Staphylococcus aureus. Mol Microbiol. 1999 Jul;33(2):307–16.
Rechtin, T. M., et al. “Characterization of the SarA virulence gene regulator of Staphylococcus aureus.Mol Microbiol, vol. 33, no. 2, July 1999, pp. 307–16. Pubmed, doi:10.1046/j.1365-2958.1999.01474.x.
Rechtin TM, Gillaspy AF, Schumacher MA, Brennan RG, Smeltzer MS, Hurlburt BK. Characterization of the SarA virulence gene regulator of Staphylococcus aureus. Mol Microbiol. 1999 Jul;33(2):307–316.
Journal cover image

Published In

Mol Microbiol

DOI

ISSN

0950-382X

Publication Date

July 1999

Volume

33

Issue

2

Start / End Page

307 / 316

Location

England

Related Subject Headings

  • Trans-Activators
  • Staphylococcus aureus
  • Protein Conformation
  • Molecular Sequence Data
  • Microbiology
  • Gene Expression Regulation, Bacterial
  • Dimerization
  • DNA Footprinting
  • Circular Dichroism
  • Base Sequence