Functional Characterization and Biological Significance of Yersinia pestis Lipopolysaccharide Biosynthesis Genes

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S. V. Dentovskaya¹, A. P. Anisimov¹, A. N. Kondakova^2,3, B. Lindner³, O. V. Bystrova², T. E. Svetoch¹, R. Z. Shaikhutdinova¹, S. A. Ivanov¹, I. V. Bakhteeva¹, G. M. Titareva¹, and Yu. A. Knirel²

¹State Research Center for Applied Microbiology and Biotechnology, 142279 Obolensk, Moscow Region, Russia; fax: (4967) 360-061; E-mail: anisimov@obolensk.org

²Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky pr. 47, 119991 Moscow, Russia; fax: (499) 135-5328; E-mail: annakond@gmail.com

³Research Center Borstel, Center for Medicine and Biosciences, D-23485 Borstel, Parkallee 22, Germany; E-mail: blindner@fz-borstel.de

^* To whom correspondence should be addressed.

Received November 27, 2010; Revision received February 4, 2011

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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 LD₅₀ 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.

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KEY WORDS: Yersinia pestis, lipopolysaccharide biosynthesis, virulence, serum resistance, antimicrobial-peptide resistance, fibrinolytic activity, coagulase activity

DOI: 10.1134/S0006297911070121

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