Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence

Killikelly, April; Jakoncic, Jean; Benson, Meredith A.; Ohneck, Elizabeth A.; Sampson, Jared M.; Spurrier, Brett; Torres, Victoer J.; Kong, Xian -Peng

doi:10.1128/JB.02317-14

Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence

Journal Article · Mon Oct 20 00:00:00 EDT 2014 · Journal of Bacteriology

DOI:https://doi.org/10.1128/JB.02317-14· OSTI ID:1174114

Killikelly, April ^[1]; Jakoncic, Jean ^[2]; Benson, Meredith A. ^[3]; Ohneck, Elizabeth A. ^[3]; Sampson, Jared M. ^[1]; Spurrier, Brett ^[1]; Torres, Victoer J. ^[3]; Kong, Xian -Peng ^[1]

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.

View Accepted Manuscript (DOE)

Research Organization:: Brookhaven National Laboratory (BNL), Upton, NY (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

OSTI ID:: 1174114

Alternate ID(s):: OSTI ID: 1229412

Report Number(s):: BNL--107535-2015-JA

Journal Information:: Journal of Bacteriology, Journal Name: Journal of Bacteriology Journal Issue: 1 Vol. 197; ISSN 0021-9193

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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36 MATERIALS SCIENCE
Staphylococcus aureus
crystal structure
repressor of toxin (Rot)
site-directed mutagenesis
winged-helix-turn-helix (WHTH)

Structure-based functional characterization of repressor of toxin (Rot), a central regulator of staphylococcus aureus virulence

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References (54)

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