The heme sensing response regulator HssR in Staphylococcus aureus but not the homologous RR23 in Listeria monocytogenes modulates susceptibility to the antimicrobial peptide plectasin

<p>Abstract</p> <p>Background</p> <p>Host defence peptides (HDPs), also known as antimicrobial peptides (AMPs), have emerged as potential new therapeutics and their antimicrobial spectrum covers a wide range of target organisms. However, the mode of action and the genetics behind the bacterial response to HDPs is incompletely understood and such knowledge is required to evaluate their potential as antimicrobial therapeutics. Plectasin is a recently discovered HDP active against Gram-positive bacteria with the human pathogen, <it>Staphylococcus aureus </it>(<it>S. aureus</it>) being highly susceptible and the food borne pathogen, <it>Listeria monocytogenes </it>(<it>L. monocytogenes</it>) being less sensitive. In the present study we aimed to use transposon mutagenesis to determine the genetic basis for <it>S. aureus </it>and <it>L. monocytogenes </it>susceptibility to plectasin.</p> <p>Results</p> <p>In order to identify genes that provide susceptibility to plectasin we constructed bacterial transposon mutant libraries of <it>S. aureus </it>NCTC8325-4 and <it>L. monocytogenes </it>4446 and screened for increased resistance to the peptide. No resistant mutants arose when <it>L. monocytogenes </it>was screened on plates containing 5 and 10 fold Minimal Inhibitory Concentration (MIC) of plectasin. However, in <it>S. aureus</it>, four mutants with insertion in the heme response regulator (<it>hssR</it>) were 2-4 fold more resistant to plectasin as compared to the wild type. The <it>hssR </it>mutation also enhanced resistance to the plectasin-like defensin eurocin, but not to other classes of HDPs or to other stressors tested. Addition of plectasin did not influence the expression of <it>hssR </it>or <it>hrtA</it>, a gene regulated by HssR. The genome of <it>L. monocytogenes </it>LO28 encodes a putative HssR homologue, RR23 (in <it>L. monocytogenes </it>EGD-e lmo2583) with 48% identity to the <it>S. aureus </it>HssR, but a mutation in the <it>rr23 </it>gene did not change the susceptibility of <it>L. monocytogenes </it>to plectasin.</p> <p>Conclusions</p> <p><it>S. aureus </it>HssR, but not the homologue RR23 from <it>L. monocytogenes</it>, provides susceptibility to the defensins plectasin and eurocin. Our data suggest that a functional difference between response regulators HssR and RR23 is responsible for the difference in plectasin susceptibility observed between <it>S. aureus </it>and <it>L. monocytogenes</it>.</p