Identification and characterization of Klebsiella pneumoniae two lpxL lipid A late acyltransferases and their role in virulence.

Grant Mills, Amy Dumigan, Jose A. Bengoechea, Timothy Kidd

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18 Citations (Scopus)
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Abstract

Klebsiella pneumoniae causes a wide range of infections, from urinary tract infections to pneumonia. The lipopolysaccharide is a virulence factor of this pathogen, although there are gaps in our understanding of its biosynthesis. Here we report the characterization of K. pneumoniae LpxL which encodes one of the enzymes responsible for late secondary acylation of immature lipid A molecules. Analysis of the available K. pneumoniae genomes revealed that this pathogen encodes two orthologues of Escherichia coli LpxL. Using genetic methods and mass spectrometry, we demonstrate that LpxL1 catalyses the addition of laureate and LpxL2 the addition of myristate. Both enzymes acylated E. coli lipid A whereas only LpxL2 mediated K. pneumoniae lipid A acylation. We show that LpxL1 is negatively regulated by the two-component system PhoPQ. The lipid A produced by lpxL2 mutant lacked the 2-hydroxymyristate, palmitate, and 4-aminoarabinose decorations found in the lipid A synthesized by the wild type. The lack of 2-hydroxymiristate was expected since LpxO modifies the myristate transferred by LpxL2 to the lipid A. The absence of the other two decorations is most likely caused by downregulation of phoPQ and pmrAB expression. LpxL2-dependent lipid A acylation protects Klebsiella from polymyxins, mediates resistance to phagocytosis, limits the activation of inflammatory responses by macrophages, and is required for pathogen survival in the waxmoth Galleria mellonella. Our findings indicate that LpxL2 contribution to virulence is dependent on LpxO-mediated hydroxylation of the LpxL2-transferred myristate. Our studies suggest that LpxL2 might be a candidate target for developing anti K. pneumoniae drugs.
Original languageEnglish
JournalInfection and Immunity
Early online date26 Jun 2017
DOIs
Publication statusEarly online date - 26 Jun 2017

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