Cationic antimicrobial peptides serve as activation signals for the Salmonella Typhimurium PhoPQ and PmrAB regulons in vitro and in vivo.
Salmonella enterica serovar Typhimurium uses two-component regulatory systems (TCRSs) to respond to environmental stimuli. Upon infection, the TCRSs PhoP-PhoQ (PhoPQ) and PmrA-PmrB (PmrAB) are activated by environmental signals detected in the lumen of the intestine and within host cells. TCRS-mediated gene expression leads to upregulation of genes involved in lipopolysaccharide (LPS) modification and cationic antimicrobial peptide (CAMP) resistance. This research expands on previous studies which have shown that CAMPs can activate Salmonella TCRSs in vitro. The focus of this work was to determine if CAMPs can act as environmental signals for PhoPQ- and PmrAB-mediated gene expression in vitro, during infection of macrophages and in a mouse model of infection. Monitoring of PhoPQ and PmrAB activation using recombinase-based in vivo expression technology (RIVET), alkaline phosphtase and β-galactosidase reporter fusion constructs demonstrated that S. Typhimurium PhoQ can sense CAMPs in vitro. In mouse macrophages, the cathelecidin CRAMP does not activate the PhoPQ regulon. Acidification of the Salmonella-containing vacuole activates PhoP- and PmrA-regulated loci but blocking acidification still does not reveal a role for CRAMP in TCRS activation in mouse macrophages. However, assays performed in susceptible wild type (WT), CRAMP knockout (KO), and matrilysin (a metalloproteinase necessary for activating murine α-defensins) KO mice suggest CRAMP, but not α-defensins, serve as a putative direct TCRS activation signal in the mouse intestine. These studies provide a better understanding of the in vivo environments that result in activation of these virulence-associated TCRSs.
Department of Microbial Infection and Immunity, Center for Microbial Interface Biology, The Ohio State University, Columbus OH, USA.
Susan M
SM
Richards
Kristi L
KL
Strandberg
Megan
M
Conroy
John S
JS
Gunn
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000096:biosynthesis*; D004195:Disease Models, Animal; D005260:Female; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D017930:Genes, Reporter; D008264:Macrophages / Q000276:immunology / Q000382:microbiology; D051379:Mice; D018085:Regulon; D012481:Salmonella Infections, Animal / Q000276:immunology / Q000382:microbiology; D012486:Salmonella typhimurium / Q000187:drug effects* / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis*; D023084:alpha-Defensins / Q000378:metabolism; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics; D054804:Cathelicidins
D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; C079792:PhoQ protein, Bacteria; C409039:PmrB protein, bacteria; D014157:Transcription Factors; D023084:alpha-Defensins; C081902:pmrA protein, Bacteria; C061064:PhoP protein, Bacteria; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase; D054804:Cathelicidins