Identification of a Small Molecule Inhibitor of Bacterial AraC Family Activators
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Issue Date
2013-01-30Author
Skredenske, Jeffrey M.
Koppolu, Veerendra
Kolin, Ana
Deng, James
Kettle, Bria Collette
Taylor, Byron
Egan, Susan M.
Publisher
SAGE Publications
Type
Article
Article Version
Scholarly/refereed, author accepted manuscript
Rights
Copyright SAGE Publications
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Show full item recordAbstract
Protein members of the AraC family of bacterial transcriptional activators have great promise as targets for the development of novel antibacterial agents. Here, we describe an in vivo high throughput screen to identify inhibitors of the AraC family activator protein RhaS. The screen used two E. coli reporter fusions; one to identify potential RhaS inhibitors, and a second to eliminate non-specific inhibitors from consideration. One compound with excellent selectivity, OSSL_051168, was chosen for further study. OSSL_051168 inhibited in vivo transcription activation by the RhaS DNA-binding domain to the same extent as the full-length protein, indicating that this domain was the target of its inhibition. Growth curves showed that OSSL_051168 did not impact bacterial cell growth at the concentrations used in this study. In vitro DNA binding assays with purified protein suggest that OSSL_051168 inhibits DNA binding by RhaS. In addition, we found that it inhibits DNA binding by a second AraC family protein, RhaR, which shares 30% amino acid identity with RhaS. OSSL_051168 did not have a significant impact on DNA binding by the non-AraC family proteins CRP and LacI, suggesting that the inhibition is likely specific for RhaS, RhaR, and possibly additional AraC family activator proteins.
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Citation
Skredenske, Jeff M., Veerendra Koppolu, Ana Kolin, James Deng, Bria Kettle, Byron Taylor, and Susan M. Egan. "Identification of a Small-Molecule Inhibitor of Bacterial AraC Family Activators." Journal of Biomolecular Screening 18.5 (2013): 588-98.
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