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Journal of Bacteriology, December 2003, p. 7213-7221, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7213-7221.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

The Klebsiella pneumoniae wabG Gene: Role in Biosynthesis of the Core Lipopolysaccharide and Virulence

Luis Izquierdo,1 Núria Coderch,2 Nuria Piqué,2 Emiliano Bedini,3 Maria Michela Corsaro,3 Susana Merino,1 Sandra Fresno,1 Juan M. Tomás,1* and Miguel Regué2

Departamento de Microbiología y Parasitología Sanitarias, División de Ciencias de la Salud, Facultad de Farmacia,2 Departamento de Microbiología, Facultad de Biología,Universidad de Barcelona, Barcelona, Spain,1 Dipartimento di Chimica e Biochimica, Università Federico II di Napoli, Naples, Italy3

Received 16 September 2003/ Accepted 30 September 2003

To determine the function of the wabG gene in the biosynthesis of the core lipopolysaccharide (LPS) of Klebsiella pneumoniae, we constructed wabG nonpolar mutants. Data obtained from the comparative chemical and structural analysis of LPS samples obtained from the wild type, the mutant strain, and the complemented mutant demonstrated that the wabG gene is involved in attachment to {alpha}-L-glycero-D-manno-heptopyranose II (L,D-HeppII) at the O-3 position of an {alpha}-D-galactopyranosyluronic acid ({alpha}-D-GalAp) residue. K. pneumoniae nonpolar wabG mutants were devoid of the cell-attached capsular polysaccharide but were still able to produce capsular polysaccharide. Similar results were obtained with K. pneumoniae nonpolar waaC and waaF mutants, which produce shorter LPS core molecules than do wabG mutants. Other outer core K. pneumoniae nonpolar mutants in the waa gene cluster were encapsulated. K. pneumoniae waaC, waaF, and wabG mutants were avirulent when tested in different animal models. Furthermore, these mutants were more sensitive to some hydrophobic compounds than the wild-type strains. All these characteristics were rescued by reintroduction of the waaC, waaF, and wabG genes from K. pneumoniae.

* Corresponding author. Mailing address: Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain. Phone: 34-402-4496. Fax: 34-3402-4498. E-mail: juant{at}porthos.bio.ub.es.

Journal of Bacteriology, December 2003, p. 7213-7221, Vol. 185, No. 24
0021-9193/03/$08.00+0     DOI: 10.1128/JB.185.24.7213-7221.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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