Abstract
BackgroundThe rising drug resistance in pathogenic bacteria and inefficiency of current antibiotics to meet clinical requirements has augmented the need to establish new and innovative approaches for antibacterial drug discovery involving identification of novel antibacterial targets and inhibitors. Being obligatory for bacterial growth, essential gene products are considered vital as drug targets. The bacterial protein YidC is highly conserved among pathogens and is essential for membrane protein insertion due to which it holds immense potential as a promising target for antibacterial therapy.Methods/Principal FindingsThe aim of this study was to explore the feasibility and efficacy of expressed antisense-mediated gene silencing for specific downregulation of yidC in Escherichia coli. We induced RNA silencing of yidC which resulted in impaired growth of the host cells. This was followed by a search for antibacterial compounds sensitizing the YidC depleted cells as they may act as inhibitors of the essential protein or its products. The present findings affirm that reduction of YidC synthesis results in bacterial growth retardation, which warrants the use of this enzyme as a viable target in search of novel antibacterial agents. Moreover, yidC antisense expression in E. coli resulted in sensitization to antibacterial essential oils eugenol and carvacrol. Fractional Inhibitory Concentration Indices (FICIs) point towards high level of synergy between yidC silencing and eugenol/carvacrol treatment. Finally, as there are no known YidC inhibitors, the RNA silencing approach applied in this study put forward rapid means to screen novel potential YidC inhibitors.Conclusions/SignificanceThe present results suggest that YidC is a promising candidate target for screening antibacterial agents. High level of synergy reported here between yidC silencing and eugenol/carvacrol treatment is indicative of a potential antibacterial therapy. This is the first report indicating that the essential gene yidC is a therapeutic target of the antibacterial essential oils eugenol and carvacrol in E. coli.
Highlights
Evolution of multidrug resistance in bacterial pathogens has led to a situation in which there are diminutive or no treatment options for infections with certain pathogens
AsRNA approach has been used to elaborate the mechanism of action of natural products in E. coli [16], there is a dearth of published reports describing the application of this approach for screening target specific antibacterial agents in Gram-negative bacteria
The growth curves for different levels of yidC and murA antisense RNA (asRNA) expression are presented in figure 2 which shows that silencing of yidC and murA lowered the final optical density at 600 nm (OD600) of E. coli cells
Summary
Evolution of multidrug resistance in bacterial pathogens has led to a situation in which there are diminutive or no treatment options for infections with certain pathogens. Regardless of the rise of resistant pathogens, the pace of new antibiotic approvals is falling [1,5,6]. Over the past few decades, in order to solve the problem of drug resistance, existing antibiotics are being chemically modified [1]. This has led to relatively narrow diversity of compound classes aiming at limited number of targets [7]. The rising drug resistance in pathogenic bacteria and inefficiency of current antibiotics to meet clinical requirements has augmented the need to establish new and innovative approaches for antibacterial drug discovery involving identification of novel antibacterial targets and inhibitors. The bacterial protein YidC is highly conserved among pathogens and is essential for membrane protein insertion due to which it holds immense potential as a promising target for antibacterial therapy
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