Identification of antimicrobial targets using a comprehensive genomic approach

Dezhong Yin, Brian Fox, Michael L. Lonetto, Mark R. Etherton, David J. Payne, David J. Holmes, Martin Rosenberg, Yinduo Ji

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

Regulated antisense RNA enables the construction of a defined set of conditional growth-defective/lethal strains. In this study, we expanded the regulated antisense RNA interference technology and developed a high-throughput screening strategy to identify the potential drug targets of novel antimicrobials. To prove this concept, the specific antisense sublibrary of different essential open reading frames were pooled in the presence of an inducer, and treated with or without sublethal levels of mupirocin, triclosan, or gentamicin. Antisense RNA-expressing strains that were sensitized for increased susceptibility to the antibiotics were selectively detected via DNA subtractive hybridization, microarray, and whole-cell analyses. No strain was identified as supersensitive to gentamicin because there was no target-specific antisense strain in this sublibrary. In contrast, strains expressing antisense to isoleucine tRNA synthetase (ileS) and enoyl-[acyl-carrier-protein] reductase (fabl) were specifically identified as having increased susceptibility to mupirocin and triclosan, respectively. These results demonstrated that ileS and fabl antisense strains showed significant increases of susceptibility only to their specific inhibitors. This data demonstrates that a regulated antisense RNA expression library provides an effective tool to assist in the identification of potential targets for novel antibacterial agents.

Original languageEnglish (US)
Pages (from-to)101-113
Number of pages13
JournalPharmacogenomics
Volume5
Issue number1
DOIs
StatePublished - Jan 1 2004

Keywords

  • Antibiotics
  • Antisense RNA
  • DNA microarray
  • DNA substractive hybridization
  • Mode of action
  • Staphylococcus aureus

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