High-Throughput Screening for Inhibitors of Wall Teichoic Acid Biosynthesis in Staphylococcus aureus
The world is heading toward a dangerous post-antibiotic era where antibiotics fail to treat infections. Staphylococcus aureus is the leading cause of healthcare-associated infections worldwide, and an ever-increasing percentage of them are methicillin-resistant (MRSA). New strategies are urgently needed to combat this pathogen. Wall teichoic acids (WTA) in S. aureus are polyribitol phosphate polymers that play important roles in virulence and resistance to β-lactam antibiotics. Here, we describe a high-throughput whole-cell screening platform for inhibitors targeting WTA biosynthesis. This platform takes advantage of the unique dispensability patterns of genes encoding WTA biosynthesis. We further describe follow-up dose-response assays to identify WTA inhibitors among the primary bioactives. WTA inhibitors offer an exciting opportunity for the development of novel antibacterial leads of unique mechanism in the fight against drug-resistant staphylococcal infections.
Key wordsSmall-molecule screening Small-molecule library Wall teichoic acid Antagonism screen Antibiotic Antibacterial Staphylococcus aureus
The authors would like to thank Dr. Maya Farha for her critical reading of the manuscript. Wall teichoic acid related research in the authors’ laboratory was supported by grants from the Canadian Institutes of Health Research and from the Canadian glycomics network (GlycoNet) and a Tier I Canada Research Chair award to E.D.B. O.M.E. was supported by a Michael G. DeGroote Fellowship Award in Basic Biomedical Science.
- 1.WHO (2014) Antimicrobial resistance: global report on surveillance. World Health Organization reports. World Health OrganizationGoogle Scholar
- 6.WHO (2017) Prioritization of pathogens to guide discovery, research and development of new antibiotics for drug-resistant bacterial infections including tuberculosis. World Health Organization, GenevaGoogle Scholar
- 8.The Canadian Cystic Fibrosis Registry 2013 Annual Report (2015). Cystic Fibrosis CanadaGoogle Scholar
- 10.D'Elia MA, Henderson JA, Beveridge TJ, Heinrichs DE, Brown ED (2009) The N-acetylmannosamine transferase catalyzes the first committed step of teichoic acid assembly in Bacillus subtilis and Staphylococcus aureus. J Bacteriol 191(12):4030–4034. https://doi.org/10.1128/JB.00611-08CrossRefPubMedPubMedCentralGoogle Scholar
- 12.D'Elia MA, Pereira MP, Chung YS, Zhao W, Chau A, Kenney TJ, Sulavik MC, Black TA, Brown ED (2006) Lesions in teichoic acid biosynthesis in Staphylococcus aureus lead to a lethal gain of function in the otherwise dispensable pathway. J Bacteriol 188(12):4183–4189. https://doi.org/10.1128/JB.00197-06CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Campbell J, Singh AK, Santa Maria JP Jr, Kim Y, Brown S, Swoboda JG, Mylonakis E, Wilkinson BJ, Walker S (2011) Synthetic lethal compound combinations reveal a fundamental connection between wall teichoic acid and peptidoglycan biosyntheses in Staphylococcus aureus. ACS Chem Biol 6(1):106–116. https://doi.org/10.1021/cb100269fCrossRefPubMedGoogle Scholar
- 14.Weidenmaier C, Kokai-Kun JF, Kristian SA, Chanturiya T, Kalbacher H, Gross M, Nicholson G, Neumeister B, Mond JJ, Peschel A (2004) Role of teichoic acids in Staphylococcus aureus nasal colonization, a major risk factor in nosocomial infections. Nat Med 10(3):243–245. https://doi.org/10.1038/nm991CrossRefPubMedGoogle Scholar
- 15.Farha MA, Leung A, Sewell EW, D'Elia MA, Allison SE, Ejim L, Pereira PM, Pinho MG, Wright GD, Brown ED (2013) Inhibition of WTA synthesis blocks the cooperative action of PBPs and sensitizes MRSA to beta-lactams. ACS Chem Biol 8(1):226–233. https://doi.org/10.1021/cb300413mCrossRefPubMedGoogle Scholar
- 16.Campbell J, Singh AK, Swoboda JG, Gilmore MS, Wilkinson BJ, Walker S (2012) An antibiotic that inhibits a late step in wall teichoic acid biosynthesis induces the cell wall stress stimulon in Staphylococcus aureus. Antimicrob Agents Chemother 56(4):1810–1820. https://doi.org/10.1128/AAC.05938-11CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Wang H, Gill CJ, Lee SH, Mann P, Zuck P, Meredith TC, Murgolo N, She X, Kales S, Liang L, Liu J, Wu J, Santa Maria J, Su J, Pan J, Hailey J, McGuinness D, Tan CM, Flattery A, Walker S, Black T, Roemer T (2013) Discovery of wall teichoic acid inhibitors as potential anti-MRSA beta-lactam combination agents. Chem Biol 20(2):272–284. https://doi.org/10.1016/j.chembiol.2012.11.013CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Farha MA, Czarny TL, Myers CL, Worrall LJ, French S, Conrady DG, Wang Y, Oldfield E, Strynadka NC, Brown ED (2015) Antagonism screen for inhibitors of bacterial cell wall biogenesis uncovers an inhibitor of undecaprenyl diphosphate synthase. Proc Natl Acad Sci U S A 112(35):11048–11053. https://doi.org/10.1073/pnas.1511751112CrossRefPubMedPubMedCentralGoogle Scholar
- 22.CLSI (2012) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard—Ninth Edition, CLSI document M07-A9. Clinical and Laboratory Standards Institute, Wayne, PAGoogle Scholar