Search

Background Fitness costs imposed on bacteria by antibiotic resistance mechanisms are believed to hamper their dissemination. The scale of these costs is highly variable. Some, including resistance of Staphylococcus aureus to the clinically important antibiotic mupirocin, have been reported as being cost-free, which suggests that there are few barriers preventing their global spread. However, this is not supported by surveillance data in healthy communities, which indicate that this resistance mechanism is relatively unsuccessful. Results Epistasis analysis on two collections of MRSA provides an explanation for this discord, where the mupirocin resistance-conferring mutation of the ileS gene appears to affect the levels of toxins produced by S. aureus when combined with specific polymorphisms at other loci. Proteomic analysis demonstrates that the activity of the secretory apparatus of the PSM family of toxins is affected by mupirocin resistance. As an energetically costly activity, this reduction in toxicity masks the fitness costs associated with this resistance mutation, a cost that becomes apparent when toxin production becomes necessary. This hidden fitness cost provides a likely explanation for why this mupirocin-resistance mechanism is not more prevalent, given the widespread use of this antibiotic. Conclusions With dwindling pools of antibiotics available for use, information on the fitness consequences of the acquisition of resistance may need to be considered when designing antibiotic prescribing policies. However, this study suggests there are levels of depth that we do not understand, and that holistic, surveillance and functional genomics approaches are required to gain this crucial information. Electronic supplementary material The online version of this article (10.1186/s13059-018-1469-2) contains supplementary material, which is available to authorized users.

Strains of the Gram-positive pathogen Staphylococcus aureus can be divided into four quorum sensing (QS) groups. Membership of each group is defined by the amino acid sequence of the autoinducing peptide (AIP) QS signal molecule that is encoded within the agrBDCA genetic locus and specifically within agrD. This chapter describes the use of simple, in-cell, lux-based, bio-reporters that can be used to identify/confirm the specific agr group to which a particular S. aureus isolate belongs, as well as to assess the timing and quantity of AIP produced.

Understanding how multi-drug resistant pathogens evolve is key to identifying means of curtailing their further emergence and dissemination. Fitness costs imposed on bacteria by resistance mechanisms are believed to hamper their dissemination in an antibiotic free environment, however, some have been reported to have little or no cost, which suggests there are few barriers preventing their global spread. One such apparently cost-free resistance mechanism acquired by the major human pathogenStaphylococcus aureusis to the clinically important antibiotic mupirocin, which is mediated by mutation of the highly-conserved and essential isoleucyl-tRNA synthethase (ileS) gene. In Genome Wide Association Studies (GWAS) on two genetically and geographically distinct MRSA lineages we have found this mutation to be associated with changes in bacterial virulence, driven through epistatic interactions with other loci. Given the potential dual effect of this mutation on both antibiotic resistance and virulence we adopted a proteomic approach and observed pleiotropic effects. This analysis revealed that the activity of the secretory apparatus of the PSM family of cytolytic toxins, the Pmt system, is affected in the mupirocin resistant mutant, which explains why it is less toxic. As an energetically costly activity, this reduction in toxicity masks the fitness costs associated with this resistance mutation, a cost that becomes apparent when toxin production is required. Given the widespread use of this antibiotic, and that this resistance often results from a single nucleotide substitution in theileSgene, these hidden fitness costs provide an explanation for why this resistance mechanism is not more prevalent. This work also demonstrates how population-based genomic analysis of virulence and antibiotic resistance can contribute to uncovering hidden features of the biology of microbial pathogens.

A series of 3-oxo-C12-HSL, tetramic acid, and tetronic acid analogues were synthesized to gain insights into the structural requirements for quorum sensing inhibition in Staphylococcus aureus. Compounds active against agr were noncompetitive inhibitors of the autoinducing peptide (AIP) activated AgrC receptor, by altering the activation efficacy of the cognate AIP-1. They appeared to act as negative allosteric modulators and are exemplified by 3-tetradecanoyltetronic acid 17, which reduced nasal cell colonization and arthritis in a murine infection model.

This chapter provides an overview on the role of bacterial channels in cell physiology. The proposed role of mechanosensitive (MS) channels is the rapid and nonspecific release of solutes from the cell in response to the generation of excessive turgor pressure. This model has now been tested and verified for E. coli. In most bacterial cells Cl- ions have not been predicted to have significant roles in cell physiology. In some halophilic bacteria Cl- is accumulated to provide salt balance during growth in very high salt concentrations. The aquaglyceroporins are pores that allow the permeation of water and small linear polyhydric molecules through the bacterial membrane. The cell faces two types of problems that relate to ammonium ions. First, there is the need to scavenge for ammonium when the concentration in the environment is low. Second, cells growing on broth may encounter a surfeit of cytoplasmic ammonium ions arising from the deamination of amino acids. In a wide range of bacterial genera the genes for AmtB and GlnK exist as an operon, and recent work suggests that the former is a pore that is used by cells to accelerate the passage of ammonium ions across the membrane and that GlnK is a specific component of the nitrogen-regulatory circuit that controls its activity. The increased understanding of mechanistic aspects of the channel selectivity and gating has advanced rapidly, but an appreciation of the role of channels in cell physiology lags well behind.