Evidence that WapB Is a 1,2-glucosyltransferase of Pseudomonas aeruginosa involved in lipopolysaccharide outer core biosynthesis

  1. Get@NRC: Evidence that WapB Is a 1,2-glucosyltransferase of Pseudomonas aeruginosa involved in lipopolysaccharide outer core biosynthesis (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1128/JB.00032-11
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Journal titleJournal of Bacteriology
Pages27082716; # of pages: 9
Subject1,2 glucosyltransferase; bacterial enzyme; bacterium lipopolysaccharide; core oligosaccharide; glucose; glucosyltransferase; oligosaccharide; unclassified drug; article; bacterial gene; biosynthesis; controlled study; enzyme structure; gene expression; genetic code; genetic complementation; mass spectrometry; nonhuman; nuclear magnetic resonance spectroscopy; priority journal; protein modification; Pseudomonas aeruginosa; serotype; terminal sequence; Western blotting; wild type; Genetic Complementation Test; Glucosyltransferases; Lipopolysaccharides; Magnetic Resonance Spectroscopy; Mass Spectrometry; Mutagenesis, Insertional; Pseudomonas aeruginosa; Pseudomonas aeruginosa
AbstractPseudomonas aeruginosa is an important opportunistic pathogen infecting debilitated individuals. One of the major virulence factors expressed by P. aeruginosa is lipopolysaccharide (LPS), which is composed of lipid A, core oligosaccharide (OS), and O-antigen polysaccharide. The core OS is divided into inner and outer regions. Although the structure of the outer core OS has been elucidated, the functions and mechanisms of the glycosyltransferases involved in core OS biogenesis are currently unknown. Here, we show that a previously uncharacterized gene, pa1014, is involved in outer core biosynthesis, and we propose to rename this gene wapB. We constructed a chromosomal mutant, wapB::Gm, in a PAO1 (O5 serotype) strain background. Characterization of the LPS from the mutant by Western immunoblotting showed a lack of reactivity to PAO1 outer core-specific monoclonal antibody (MAb) 5c-101. The chemical structure of the core OS of the wapB mutant was elucidated using nuclear magnetic resonance spectroscopy and mass spectrometry techniques and revealed that the core OS of the wapB mutant lacked the terminal β-1,2-linked-D-glucose residue. Complementation of the mutant with wapB in trans restored the core structure to one that is identical to that of the wild type. Eleven of the 20 P. aeruginosa International Antigenic Typing Scheme (IATS) serotypes produce LPSs that lack the terminal D-glucose residue (Glc IV). Interestingly, expressing wapB in each of these 11 serotypes modifies each of their outer core OS structures, which became reactive to MAb 5c-101 in Western immunoblotting, suggesting the presence of a terminal D-glucose in these core OS structures. Our results strongly suggested that wapB encodes a 1,2-glucosyltransferase. © 2011, American Society for Microbiology.
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AffiliationNational Research Council Canada (NRC-CNRC); NRC Institute for Biological Sciences (IBS-ISB)
Peer reviewedYes
NPARC number21271084
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Record identifier1ffc7d91-53e0-4a97-a79b-c4be8d122a3c
Record created2014-03-24
Record modified2016-05-09
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