pmrA(Con) confers pmrHFIJKL-dependent EGTA and polymyxin resistance on msbB Salmonella by decorating lipid A with phosphoethanolamine.
Mutations in pmrA were recombined into Salmonella strain ATCC 14028 msbB to determine if pmrA-regulated modifications of lipopolysaccharide could suppress msbB growth defects. A mutation that functions to constitutively activate pmrA [pmrA(Con)] suppresses msbB growth defects on EGTA-containing media. Lipid A structural analysis showed that Salmonella msbB pmrA(Con) strains, compared to Salmonella msbB strains, have increased amounts of palmitate and phosphoethanolamine but no aminoarabinose addition, suggesting that aminoarabinose is not incorporated into msbB lipid A. Surprisingly, loss-of-function mutations in the aminoarabinose biosynthetic genes restored EGTA and polymyxin sensitivity to Salmonella msbB pmrA(Con) strains. These blocks in aminoarabinose biosynthesis also prevented lipid A phosphoethanolamine incorporation and reduced the levels of palmitate addition, indicating previously unknown roles for the aminoarabinose biosynthetic enzymes. Lipid A structural analysis of the EGTA- and polymyxin-resistant triple mutant msbB pmrA(Con) pagP::Tn10, which contains phosphoethanolamine but no palmitoylated lipid A, suggests that phosphoethanolamine addition is sufficient to confer EGTA and polymyxin resistance on Salmonella msbB strains. Additionally, palmitoylated lipid A was observed only in wild-type Salmonella grown in the presence of salt in rich media. Thus, we correlate EGTA resistance and polymyxin resistance with phosphoethanolamine-decorated lipid A and demonstrate that the aminoarabinose biosynthetic proteins play an essential role in lipid A phosphoethanolamine addition and affect lipid A palmitate addition in Salmonella msbB strains.
Murray
Sean R
SR
Department of Biology, Yale University, New Haven, Connecticut, USA.
Ernst
Robert K
RK
Bermudes
David
D
Miller
Samuel I
SI
Low
K Brooks
KB
D001089:Arabinose / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002849:Chromatography, Gas; D002855:Chromatography, Thin Layer; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004533:Egtazic Acid / Q000494:pharmacology*; D004983:Ethanolamines / Q000737:chemistry* / Q000378:metabolism; D008050:Lipid A / Q000737:chemistry* / Q000378:metabolism; D050356:Lipid Metabolism / Q000187:drug effects; D050296:Microbial Viability / Q000187:drug effects / Q000235:genetics; D015394:Molecular Structure; D009154:Mutation; D019308:Palmitic Acid / Q000737:chemistry / Q000378:metabolism; D011113:Polymyxins / Q000494:pharmacology*; D012475:Salmonella / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D012492:Salts / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001426:Bacterial Proteins; D004983:Ethanolamines; D008050:Lipid A; D011113:Polymyxins; D012492:Salts; C016982:aminoarabinose; C081902:pmrA protein, Bacteria; D019308:Palmitic Acid; D004533:Egtazic Acid; C005448:phosphorylethanolamine; D001089:Arabinose