Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence
Abstract
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.more »
- Authors:
-
- 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.
- Publication Date:
- Research Org.:
- Brookhaven National Laboratory (BNL), Upton, NY (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1174114
- Report Number(s):
- BNL-107535-2015-JA
Journal ID: ISSN 0021-9193; R&D Project: LS001
- Grant/Contract Number:
- SC00112704
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Bacteriology
- Additional Journal Information:
- Journal Volume: 197; Journal Issue: 1; Journal ID: ISSN 0021-9193
- Publisher:
- American Society for Microbiology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 36 MATERIALS SCIENCE; Staphylococcus aureus; repressor of toxin (Rot); crystal structure; winged-helix-turn-helix (WHTH); site-directed mutagenesis
Citation Formats
Killikelly, April, Jakoncic, Jean, Benson, Meredith A., Ohneck, Elizabeth A., Sampson, Jared M., Spurrier, Brett, Torres, Victoer J., and Kong, Xian -Peng. Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence. United States: N. p., 2014.
Web. doi:10.1128/JB.02317-14.
Killikelly, April, Jakoncic, Jean, Benson, Meredith A., Ohneck, Elizabeth A., Sampson, Jared M., Spurrier, Brett, Torres, Victoer J., & Kong, Xian -Peng. Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence. United States. https://doi.org/10.1128/JB.02317-14
Killikelly, April, Jakoncic, Jean, Benson, Meredith A., Ohneck, Elizabeth A., Sampson, Jared M., Spurrier, Brett, Torres, Victoer J., and Kong, Xian -Peng. Mon .
"Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence". United States. https://doi.org/10.1128/JB.02317-14. https://www.osti.gov/servlets/purl/1174114.
@article{osti_1174114,
title = {Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence},
author = {Killikelly, April and Jakoncic, Jean and Benson, Meredith A. and Ohneck, Elizabeth A. and Sampson, Jared M. and Spurrier, Brett and Torres, Victoer J. and Kong, Xian -Peng},
abstractNote = {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.},
doi = {10.1128/JB.02317-14},
journal = {Journal of Bacteriology},
number = 1,
volume = 197,
place = {United States},
year = {Mon Oct 20 00:00:00 EDT 2014},
month = {Mon Oct 20 00:00:00 EDT 2014}
}
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