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J Biol Chem, Vol. 273, Issue 41, 26310-26316, October 9, 1998
From the The waaY, waaQ, and
waaP genes are located in the central operon of the
waa (formerly rfa) locus on the chromosome of
Escherichia coli. This locus contains genes whose products
are involved in the assembly of the core region of the
lipopolysaccharide molecule. In the R1 core prototype strain, E. coli F470, there are nine genes in this operon, and all but
waaY, waaQ, and waaP have been assigned function. In this study, the waaY,
waaQ, and waaP genes were independently mutated
by insertion of a non-polar antibiotic resistance cassette, and the
structures of the resulting mutant core oligosaccharides were
determined by chemical analyses and phosphorus-nuclear magnetic
resonance spectroscopy. All three of these mutations were shown to
affect the modification of the heptose region of the core, a region
whose structure is critical to outer membrane stability. Mutation of
waaY resulted in a core oligosaccharide devoid of phosphate
on HepII. Mutation of waaQ resulted in loss of the branch
HepIII residue on HepII and impeded the activity of WaaY. Mutation of
waaP resulted in loss of phosphoryl substituents on HepI
and obviated WaaQ and WaaY activity. Only mutation of waaP
resulted in hypersensitivity to novobiocin and sodium dodecyl sulfate,
a characteristic of deep-rough mutations.
Involvement of waaY, waaQ, and
waaP in the Modification of Escherichia coli
Lipopolysaccharide and Their Role in the Formation of a Stable Outer
Membrane
,,
,, and
Department of Microbiology, University of
Guelph, Guelph, Ontario N1G 2W1, Canada and the Institute for
Biological Sciences, National Research Council, Ottawa,
Ontario K1A OR6, Canada
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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