Roles of 3-deoxy-D-manno-2-octulosonic acid transferase from Moraxella catarrhalis in lipooligosaccharide biosynthesis and virulence.
Lipooligosaccharide (LOS), a major outer membrane component of Moraxella catarrhalis, is a possible virulence factor in the pathogenesis of human infections caused by the organism. However, information about the roles of the oligosaccharide chain from LOS in bacterial infection remains limited. Here, a kdtA gene encoding 3-deoxy-D-manno-2-octulosonic acid (Kdo) transferase, which is responsible for adding Kdo residues to the lipid A portion of the LOS, was identified by transposon mutagenesis and construction of an isogenic kdtA mutant in strain O35E. The resulting O35EkdtA mutant produced only lipid A without any core oligosaccharide, and it was viable. Physicochemical and biological analysis revealed that the mutant was susceptible to hydrophobic reagents and a hydrophilic glycopeptide and was sensitive to bactericidal activity of normal human serum. Importantly, the mutant showed decreased toxicity by the Limulus amebocyte lysate assay, reduced adherence to human epithelial cells, and enhanced clearance in lungs and nasopharynx in a mouse aerosol challenge model. These data suggest that the oligosaccharide moiety of the LOS is important for the biological activity of the LOS and the virulence capability of the bacteria in vitro and in vivo. This study may bring new insights into novel vaccines or therapeutic interventions against M. catarrhalis infections.
Peng
Daxin
D
Vaccine Research Section, National Institute on Deafness and Other Communication Disorders, Rockville, Maryland 20850, USA.
Choudhury
Biswa P
BP
Petralia
Ronald S
RS
Carlson
Russell W
RW
Gu
Xin-Xing
XX
D001422:Bacterial Adhesion; D003001:Cloning, Molecular; D008401:Gas Chromatography-Mass Spectrometry; D006367:HeLa Cells; D006801:Humans; D008070:Lipopolysaccharides / Q000032:analysis / Q000096:biosynthesis*; D001936:Moraxella catarrhalis / Q000201:enzymology / Q000254:growth & development / Q000472:pathogenicity*; D013400:Sugar Acids / Q000032:analysis; D014166:Transferases / Q000502:physiology*; D014774:Virulence
D008070:Lipopolysaccharides; D013400:Sugar Acids; C023023:lipid-linked oligosaccharides; C002532:2-keto-3-deoxyoctonate; D014166:Transferases; C019917:3-deoxy-D-manno-octulosonate transferase