The inner-core lipopolysaccharide biosynthetic waaE gene: function and genetic distribution among some Enterobacteriaceae.
To determine the function of the waaE gene in the biosynthesis of the inner-core LPS of Klebsiella pneumoniae, a waaE non-polar mutant has been constructed. Data obtained from the comparative chemical analysis of LPS samples obtained from the wild-type, the mutant strain and the complemented mutant demonstrated that the waaE gene is involved in substitution of alpha-L-glycero-D-manno-heptopyranose I (L,D-HeppI) at the O-4 position by a beta-D-glucopyranose (beta-D-Glcp) residue. In addition, DNA amplification and nucleotide sequence determination studies revealed that waaE homologues located between the waaA and coaD genes are present in clinical isolates of Enterobacteriaceae containing the structure beta-D-Glcp-(1-->4)-alpha-L,D-HeppI (K. pneumoniae, Proteus mirabilis and Yersinia enterocolitica), as well as in strains of Serratia marcescens and Enterobacter aerogenes of unknown LPS-core structures. Complementation studies using non-polar waaE mutants prove that all the waaE homologues perform the same function. Furthermore, K. pneumoniae, Ser. marcescens and P. mirabilis non-polar waaE mutants showed reduced adhesion and pathogenicity. In addition, the Ser. marcescens and P. murabilis waaE mutants showed reduced swarming motility and ability to form biofilms in vitro. All these characteristics were rescued by reintroduction of the waaE gene independently of its origin. An easy DNA amplification method to detect this gene was established, which also helps in finding the potential presence of this structural feature [beta-D-Glcp-(1-->4)-alpha-L,D-HeppI] in the inner-core LPS of Enterobacteriaceae members with unknown LPS-core structures.
Izquierdo
Luis
L
Departamento de Microbiologı́a, Facultad de Biologı́a, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain2.
Abitiu
Nihal
N
Departamento de Microbiologı́a y Parasitologı́a Sanitarias, División de Ciencias de la Salud, Facultad de Farmacia, Universidad de Barcelona, Av. Joan XXIII s/n, Barcelona 08028, Spain1.
Coderch
Núria
N
Departamento de Microbiologı́a y Parasitologı́a Sanitarias, División de Ciencias de la Salud, Facultad de Farmacia, Universidad de Barcelona, Av. Joan XXIII s/n, Barcelona 08028, Spain1.
Hita
Beatriz
B
Departamento de Microbiologı́a y Parasitologı́a Sanitarias, División de Ciencias de la Salud, Facultad de Farmacia, Universidad de Barcelona, Av. Joan XXIII s/n, Barcelona 08028, Spain1.
Merino
Susana
S
Departamento de Microbiologı́a, Facultad de Biologı́a, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain2.
Gavin
Rosalina
R
Departamento de Microbiologı́a, Facultad de Biologı́a, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain2.
Tomás
Juan M
JM
Departamento de Microbiologı́a, Facultad de Biologı́a, Universidad de Barcelona, Diagonal 645, 08071 Barcelona, Spain2.
Regué
Miguel
M
Departamento de Microbiologı́a y Parasitologı́a Sanitarias, División de Ciencias de la Salud, Facultad de Farmacia, Universidad de Barcelona, Av. Joan XXIII s/n, Barcelona 08028, Spain1.
D001426:Bacterial Proteins / Q000378:metabolism* / Q000502:physiology; D018441:Biofilms / Q000254:growth & development*; D004755:Enterobacteriaceae / Q000378:metabolism* / Q000502:physiology; D005816:Genetic Complementation Test; D005964:Glucosyltransferases; D007711:Klebsiella pneumoniae / Q000378:metabolism / Q000502:physiology; D008070:Lipopolysaccharides / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009068:Movement; D009154:Mutation; D011513:Proteus mirabilis / Q000378:metabolism / Q000502:physiology; D012706:Serratia marcescens / Q000378:metabolism / Q000502:physiology
D001426:Bacterial Proteins; D008070:Lipopolysaccharides; D005964:Glucosyltransferases; C430063:WaaE protein, Klebsiella pneumoniae