title: Functional characterization and biological significance of Yersinia pestis lipopolysaccharide biosynthesis genes.
abstract: In silico analysis of available bacterial genomes revealed the phylogenetic proximity levels of enzymes responsible for biosynthesis of lipopolysaccharide (LPS) of Yersinia pestis, the cause of plague, to homologous proteins of closely related Yersinia spp. and some other bacteria (Serratia proteamaculans, Erwinia carotovora, Burkholderia dolosa, Photorhabdus luminescens and others). Isogenic Y. pestis mutants with single or double mutations in 14 genes of LPS biosynthetic pathways were constructed by site-directed mutagenesis on the base of the virulent strain 231 and its attenuated derivative. Using high-resolution electrospray ionization mass spectrometry, the full LPS structures were elucidated in each mutant, and the sequence of monosaccharide transfers in the assembly of the LPS core was inferred. Truncation of the core decreased significantly the resistance of bacteria to normal human serum and polymyxin B, the latter probably as a result of a less efficient incorporation of 4-amino-4-deoxyarabinose into lipid A. Impairing of LPS biosynthesis resulted also in reduction of LPS-dependent enzymatic activities of plasminogen activator and elevation of LD(50) and average survival time in mice and guinea pigs infected with experimental plague. Unraveling correlations between biological properties of bacteria and particular LPS structures may help a better understanding of pathogenesis of plague and implication of appropriate genes as potential molecular targets for treatment of plague.
authors: State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russia.
authors: S V
authors: SV
authors: Dentovskaya
authors: A P
authors: AP
authors: Anisimov
authors: A N
authors: AN
authors: Kondakova
authors: B
authors: B
authors: Lindner
authors: O V
authors: OV
authors: Bystrova
authors: T E
authors: TE
authors: Svetoch
authors: R Z
authors: RZ
authors: Shaikhutdinova
authors: S A
authors: SA
authors: Ivanov
authors: I V
authors: IV
authors: Bakhteeva
authors: G M
authors: GM
authors: Titareva
authors: And Yu A
authors: AY
authors: Knirel
mesh_terms: D000606:Amino Sugars / Q000378:metabolism; D000818:Animals; D001770:Blood Bactericidal Activity; D024881:Drug Resistance, Bacterial; D005260:Female; D005798:Genes, Bacterial / Q000502:physiology*; D006168:Guinea Pigs; D006801:Humans; D008050:Lipid A / Q000096:biosynthesis; D008070:Lipopolysaccharides / Q000096:biosynthesis*; D008297:Male; D051379:Mice; D010930:Plague / Q000382:microbiology; D010960:Plasminogen Activators / Q000378:metabolism; D011112:Polymyxin B / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D014774:Virulence; D015010:Yersinia pestis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity
chemical_list: D000606:Amino Sugars; D008050:Lipid A; D008070:Lipopolysaccharides; C040134:4-amino-4-deoxyarabinose; D010960:Plasminogen Activators; D011112:Polymyxin B