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Characterization and Biological Role of the O-Polysaccharide Gene Cluster of Yersinia enterocolitica Serotype O:9

Mikael Skurnik,^1,2* Marta Biedzka-Sarek,¹ Peter S. Lübeck,^3, Tea Blom,^1, José Antonio Bengoechea,⁵ Camino Pérez-Gutiérrez,⁵ Peter Ahrens,^4, and Jeffrey Hoorfar³

Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki,¹ Helsinki University Central Hospital Laboratory Diagnostics, Helsinki, Finland,² National Food Institute,³ National Veterinary Institute, Technical University of Denmark, Copenhagen, Denmark,⁴ Unidad de Investigacion, Hospital Son Dureta, Palma Mallorca, and Program Infection and Immunity, Fundación Caubet-Cimera Illes Balears, Bunyola, Spain⁵

Received 19 April 2007/ Accepted 16 July 2007

Yersinia enterocolitica serotype O:9 is a gram-negative enteropathogen that infects animals and humans. The role of lipopolysaccharide (LPS) in Y. enterocolitica O:9 pathogenesis, however, remains unclear. The O:9 LPS consists of lipid A to which is linked the inner core oligosaccharide, serving as an attachment site for both the outer core (OC) hexasaccharide and the O-polysaccharide (OPS; a homopolymer of N-formylperosamine). In this work, we cloned the OPS gene cluster of O:9 and identified 12 genes organized into four operons upstream of the gnd gene. Ten genes were predicted to encode glycosyltransferases, the ATP-binding cassette polysaccharide translocators, or enzymes required for the biosynthesis of GDP-N-formylperosamine. The two remaining genes within the OPS gene cluster, galF and galU, were not ascribed a clear function in OPS biosynthesis; however, the latter gene appeared to be essential for O:9. The biological functions of O:9 OPS and OC were studied using isogenic mutants lacking one or both of these LPS parts. We showed that OPS and OC confer resistance to human complement and polymyxin B; the OPS effect on polymyxin B resistance could be observed only in the absence of OC.

Published ahead of print on 10 August 2007.

Present address: Bio Science and Technology, BioCentrum, Technical University of Denmark, Copenhagen, Denmark.

Present address: Molecular Cancer Biology Program, 00014 University of Helsinki, Finland.

Present address: Statens Serum Institut, Copenhagen, Denmark.

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