High-Throughput Screening for Inhibitors of Wall Teichoic Acid Biosynthesis in Staphylococcus aureus

  • Omar M. El-Halfawy
  • Eric D. BrownEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1954)


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 words

Small-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. 1.
    WHO (2014) Antimicrobial resistance: global report on surveillance. World Health Organization reports. World Health OrganizationGoogle Scholar
  2. 2.
    Li XZ, Plesiat P, Nikaido H (2015) The challenge of efflux-mediated antibiotic resistance in gram-negative bacteria. Clin Microbiol Rev 28(2):337–418. Scholar
  3. 3.
    Starosta AL, Lassak J, Jung K, Wilson DN (2014) The bacterial translation stress response. FEMS Microbiol Rev 38(6):1172–1201. Scholar
  4. 4.
    Marrakchi M, Liu X, Andreescu S (2014) Oxidative stress and antibiotic resistance in bacterial pathogens: state of the art, methodologies, and future trends. Adv Exp Med Biol 806:483–498. Scholar
  5. 5.
    de la Fuente-Nunez C, Reffuveille F, Fernandez L, Hancock RE (2013) Bacterial biofilm development as a multicellular adaptation: antibiotic resistance and new therapeutic strategies. Curr Opin Microbiol 16(5):580–589. Scholar
  6. 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
  7. 7.
    Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG Jr (2015) Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev 28(3):603–661. Scholar
  8. 8.
    The Canadian Cystic Fibrosis Registry 2013 Annual Report (2015). Cystic Fibrosis CanadaGoogle Scholar
  9. 9.
    Sewell EW, Brown ED (2014) Taking aim at wall teichoic acid synthesis: new biology and new leads for antibiotics. J Antibiot 67(1):43–51. Scholar
  10. 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. Scholar
  11. 11.
    D'Elia MA, Millar KE, Beveridge TJ, Brown ED (2006) Wall teichoic acid polymers are dispensable for cell viability in Bacillus subtilis. J Bacteriol 188(23):8313–8316. Scholar
  12. 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. Scholar
  13. 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. Scholar
  14. 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. Scholar
  15. 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. Scholar
  16. 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. Scholar
  17. 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. Scholar
  18. 18.
    Zlitni S, Blanchard JE, Brown ED (2009) High-throughput screening of model bacteria. Methods Mol Biol 486:13–27. Scholar
  19. 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. Scholar
  20. 20.
    Czarny TL, Brown ED (2016) A small-molecule screening platform for the discovery of inhibitors of undecaprenyl diphosphate synthase. ACS Infect Dis 2(7):489–499. Scholar
  21. 21.
    Lee K, Campbell J, Swoboda JG, Cuny GD, Walker S (2010) Development of improved inhibitors of wall teichoic acid biosynthesis with potent activity against Staphylococcus aureus. Bioorg Med Chem Lett 20(5):1767–1770. Scholar
  22. 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
  23. 23.
    Zhang JH, Chung TD, Oldenburg KR (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4(2):67–73. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and Biomedical SciencesMcMaster UniversityHamiltonCanada
  2. 2.Michael G. DeGroote Institute of Infectious Disease ResearchMcMaster UniversityHamiltonCanada
  3. 3.Microbiology and Immunology Department, Faculty of PharmacyAlexandria UniversityAlexandriaEgypt

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