The Pseudomonas aeruginosa algC gene encodes phosphoglucomutase, required for the synthesis of a complete lipopolysaccharide core.
We have constructed strains of Pseudomonas aeruginosa with mutations in the algC gene, previously shown to encode the enzyme phosphomannomutase. The algC mutants of a serotype O5 strain (PAO1) and a serotype O3 strain (PAC1R) did not express lipopolysaccharide (LPS) O side chains or the A-band (common antigen) polysaccharide. The migration of LPS from the algC mutant strains in Tricine-sodium dodecyl sulfate-polyacrylamide gels was similar to that of LPS from a PAO1 LPS-rough mutant, strain AK1012, and from a PAC1R LPS-rough mutant, PAC605, each previously shown to be deficient in the incorporation of glucose onto the LPS core (K. F. Jarrell and A. M. Kropinski, J. Virol. 40:411-420, 1981, and P. S. N. Rowe and P. M. Meadow, Eur. J. Biochem. 132:329-337, 1983). We show that, as expected, the algC mutant strains had no detectable phosphomannomutase activity and that neither algC strain had detectable phosphoglucomutase (PGM) activity. To confirm that the PGM activity was encoded by the algC gene, we transferred the cloned, intact P. aeruginosa algC gene to a pgm mutant of Escherichia coli and observed complementation of the pgm phenotype. Our finding that the algC gene product has PGM activity and that strains with mutations in this gene produce a truncated LPS core suggests that the synthesis of glucose 1-phosphate is necessary in the biosynthesis of the P. aeruginosa LPS core. The data presented here thus demonstrate that the algC gene is required for the synthesis of a complete LPS core in two strains with different LPS core and O side chain structures.
Coyne
M J
MJ
Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115-5899.
Russell
K S
KS
Coyle
C L
CL
Goldberg
J B
JB
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D005958:Glucosephosphates / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism*; D008360:Mannosephosphates / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D019081:O Antigens; D010733:Phosphoglucomutase / Q000235:genetics* / Q000378:metabolism; D017875:Phosphotransferases (Phosphomutases) / Q000235:genetics* / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis / Q000737:chemistry; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism
D005958:Glucosephosphates; D008070:Lipopolysaccharides; D008360:Mannosephosphates; D019081:O Antigens; D011135:Polysaccharides, Bacterial; D011994:Recombinant Proteins; C047217:mannose 1-phosphate; C031590:glucose-1-phosphate; D017875:Phosphotransferases (Phosphomutases); D010733:Phosphoglucomutase; C052799:phosphomannomutase