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
-L-glycero-D-manno-heptopyranose II
(L,D-HeppII) at the O-3 position of an
-D-galactopyranosyluronic acid
(
-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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Copyright © 2003 by the American Society for Microbiology. All rights reserved.