Synthesis of Escherichia coli O9a polysaccharide requires the participation of two domains of WbdA, a mannosyltransferase encoded within the wb* gene cluster.
WbdA (previously MtfA) is one of the mannosyltransferases encoded within the Escherichia coli O9a wb* gene cluster. It is composed of two domains of similar size, connected by an alpha-helix chain. Elimination of the C-terminal half by transposon insertion or gene deletion caused synthesis of an altered structural O-polysaccharide consisting only of alpha-1,2-linked mannose. O9a polysaccharide synthesis was restored by the C-terminal half of WbdA in trans. No membrane incorporation of mannose from GDP mannose was observed in a strain carrying only the gene for truncated WbdA. For mannose incorporation, it was necessary to introduce both wbdB and wbdC genes into the strain. Therefore, it is likely that the N-terminal half of truncated WbdA synthesizes the altered O-polysaccharide together with other mannosyltransferases which participate in the initial reactions of the O9a polysaccharide synthesis. Both N- and C-terminal domains of WbdA are required for the synthesis of the complete E. coli O9a polysaccharide. The chi sequence location between the two domains and homology plot analyses of the wbdA and the WbdA protein suggested that the wbdA gene might have arisen by fusion of two independent genes.
Kido
N
N
Biosystems, School of Informatics and Sciences, Nagoya University, Japan. j45811a@nucc.cc.nagoya-u.ac.jp
Sugiyama
T
T
Yokochi
T
T
Kobayashi
H
H
Okawa
Y
Y
D017124:Conserved Sequence / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D005798:Genes, Bacterial / Q000235:genetics; D005816:Genetic Complementation Test; D008070:Lipopolysaccharides / Q000737:chemistry; D008358:Mannose / Q000378:metabolism; D008364:Mannosyltransferases / Q000378:metabolism*; D016254:Mutagenesis, Insertional / Q000235:genetics; D019081:O Antigens / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis* / Q000737:chemistry; D017433:Protein Structure, Secondary; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
D004274:DNA, Recombinant; D008070:Lipopolysaccharides; D019081:O Antigens; D011135:Polysaccharides, Bacterial; D008364:Mannosyltransferases; D008358:Mannose