Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence
- New York Univ. School of Medicine, New York, NY (United States). Dept. of Biochemistry and Molecular Pharmacology.
- Brookhaven National Laboratory (BNL), Upton, NY (United States). Photon Sciences Directorate.
- New York Univ. School of Medicine, New York, NY (United States). Dept. of Microbiology.
Staphylococcus aureus is responsible for a large number of diverse infections worldwide. In order to support its pathogenic lifestyle, S. aureus has to regulate the expression of virulence factors in a coordinated fashion. One of the central regulators of the S. aureus virulence regulatory networks is the transcription factor repressor of toxin (Rot). Rot plays a key role in regulating S. aureus virulence through activation or repression of promoters that control expression of a large number of critical virulence factors. However, the mechanism by which Rot mediates gene regulation has remained elusive. Here, we have determined the crystal structure of Rot and used this information to probe the contribution made by specific residues to Rot function. Rot was found to form a dimer, with each monomer harboring a winged helix-turn-helix (WHTH) DNA-binding motif. Despite an overall acidic pI, the asymmetric electrostatic charge profile suggests that Rot can orient the WHTH domain to bind DNA. Structure-based site-directed mutagenesis studies demonstrated that R91, at the tip of the wing, plays an important role in DNA binding, likely through interaction with the minor groove. We also found that Y66, predicted to bind within the major groove, contributes to Rot interaction with target promoters. Evaluation of Rot binding to different activated and repressed promoters revealed that certain mutations on Rot exhibit promoter-specific effects, suggesting for the first time that Rot differentially interacts with target promoters. As a result, this work provides insight into a precise mechanism by which Rot controls virulence factor regulation in S. aureus.
- Research Organization:
- Brookhaven National Laboratory (BNL), Upton, NY (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC)
- Grant/Contract Number:
- SC00112704
- OSTI ID:
- 1174114
- Report Number(s):
- BNL-107535-2015-JA; R&D Project: LS001
- Journal Information:
- Journal of Bacteriology, Vol. 197, Issue 1; ISSN 0021-9193
- Publisher:
- American Society for MicrobiologyCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Web of Science
Revisiting the regulation of the capsular polysaccharide biosynthesis gene cluster in Staphylococcus aureus
|
journal | July 2019 |
Staphylococcus aureus Coordinates Leukocidin Expression and Pathogenesis by Sensing Metabolic Fluxes via RpiRc
|
journal | June 2016 |
High Production of LukMF’ in Staphylococcus aureus Field Strains Is Associated with Clinical Bovine Mastitis
|
journal | May 2018 |
Revisiting the regulation of the capsular polysaccharide biosynthesis gene cluster in Staphylococcus aureus
|
posted_content | April 2019 |
Similar Records
Structure of the MecI repressor from Staphylococcus aureus in complex with the cognate DNA operator of mec
Structure of the Mecl Repressor from Staphylococcus aureus in Complex with the Cognate DNA Operator of mec