Summary
We previously demonstrated that regulated antisense RNA technology enables us to validate and identify the mode of action for some antibiotics. In this study, we have expanded the application of the regulated antisense approach to track the mode of action for a novel inhibitor of polypeptide deformylase (Pdf), which is an attractive target for the development of novel classes of antibacterial agents. We created a pdf antisense isogenic strain in Staphylococcus aureus using a TetR-regulated expression system. We demonstrated that the partial inhibition of pdf expression significantly increased the susceptibility of S. aureus to Pdf-specific inhibitor. This result provides further evidence that the TetR-regulated antisense technology is a robust tool for tracking the mode of action of novel antibacterial agents.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bandow J.E., Brotz H., Leichert L.I.O., Labischinski H., Hecker M., 2003 Proteomic approach to understanding antibiotic action Antimicrobial Agents and Chemotherapy 47:948–955
Chan P.F., O’Dwyer K., Palmer L., Ambrad J., Ingraham K., So C., Lonetto M., Biswas S., Rosenberg M., Holmes D., Zalacain M., 2003 Characterization of a novel fucose-regulated promoter (PfcsK) suitable for gene essentiality and antibacterial mode-of action studies in Streptococcus pneumoniae Journal of Bacteriology 185:2051–2058
Chang S.Y., McGary E.C., Chang S., 1989 Methionine aminopeptidase gene of Escherichia coli is essential for bacterial growth Journal of Bacteriology 171:4070–4072
Chen D.Z., Petel D.V., Hackbarth C.J., Wang W., Dreyer G., Young D.C., Margolis P.S., Wu C., Ni Z.J.T.J., White R.J., Yuan Z., 2000 Actinonin, a naturally occurring antibacterial agent, is a potent deformylase inhibitor Biochemistry 39:1256–1262
Forsyth R.A., Hselbeck R.J., Ohlesn K.L., Yamamoto R.T., Xu H., Trawick J.D., Wall D., Wang L., Brown-Driver V., Froelich J.M., Kedar G.C., King P., McCarthy M., Malone C., Misiner B., Robbins D., Tan Z., Zhu Z., Carr G., Mosca D.A., Zamudio C., Foulkes J.G., Zyskind J.W., 2002 A genome-wide strategy for the identification of essential genes in Staphylococcus aureus Molecular Microbiology 43:1387–1400
Geissendorfer M., Hillen W., 1990 Regulated expression of heterologous genes in Bacillus subtilis using the Tn10 encoded tet regulatory elements Applied Microbiology and Biotechnology 33:657–663
Guzman L.M., Belin D., Calson M.J., Beckwith J., 1995 Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter Journal of Bacteriology 177:4121–4130
Ji Y., 2002 The role of genomics in the discovery of novel targets for antibiotic therapy Pharmacogenomics 3:315–323
Ji Y., Zhang B., Van Horn S.F., Warren P., Woodnutt G., Burnham M.K.R., Rosenberg M., 2001 Identification of critical staphylococcal genes using conditional phenotypes generated by antisense RNA Science 293:2266–2269
Ji Y., Yin D., Fox B., Holmes D., Payne D., Rosenberg M., 2004 Validation of antibiotic mechanism of action by regulated antisense RNA expression in Staphylococcus aureus FEMS Microbiology Letters 231:177–184
Khodursky A.B., Peter B.J., Schmid M.B., DeRisi J., Botstein D., Brown P.O., Cozzarelli N.R., 2000 Analysis of topoisomerase function in bacterial replication fork movement: use of DNA microarrays Proceedings of the National Academy of Sciences of the USA 97:9419–9424
Schmid M.B., Kapur N., Isaacson D.R., Lindroos P., Sharpe C., 1989 Genetic analysis of temperature-sensitive lethal mutants of Salmonella typhimurium Genetics 123:625–633
Shlaes D., Projan S., Edwards J., 2004 Antibiotic discovery: state of the state ASM News 70:275–281
Stieger M., Wohlgensinger B., Kamber M., Rolf L., Keck W., 1999 Integrational plasmids for the tetracycline-regulated expression of genes in Streptococcus pneumoniae Gene 226:243–251
Yin D., Fox B., Lonetto M.L., Etherton M.R., Payne D.J., Holmes D.J., Rosenberg M., Ji Y., 2004 Identification of antimicrobial targets using a comprehensive genomic approach Pharmocogenomics 5:101–113
Zhang L., Fan F., Palmer L., Lonetto M., Petit C., Voelker L., John A., Bankosky B., Rosenberg M., McDevitt D., 2000 Regulated gene expression in Staphylococcus aureus for identifying conditional lethal phenotypes and antibiotic mode of action Gene 255:297–305
Acknowledgements
We thank Dr. Martin Burnham for his critical review and suggestions, Dr. F. Fan for strain RN4220/tetM, Dr. J. Huang for plasmid DNA pYH4, and Dr. L. Volker for providing compounds.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, J., Zheng, L. & Ji, Y. Confirmation of the mode of action of an antibacterial inhibitor using regulated antisense RNA. World J Microbiol Biotechnol 22, 299–303 (2006). https://doi.org/10.1007/s11274-005-9036-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-005-9036-x