Role of phosphoglucomutase in lipooligosaccharide biosynthesis in Neisseria gonorrhoeae.
A region of pSG30 that complements the pyocin-derived gonococcal lipooligosaccharide (LOS) mutants 1291d and 1291e was characterized by DNA sequence analysis and an open reading frame of 1,380 bases was identified that is 89% similar and 56% identical over 452 amino acids to the algC gene product from Pseudomonas aeruginosa that encodes phosphomannomutase. Enzymatic analysis of gonococcal crude protein extracts demonstrated that pSG30 encodes phosphoglucomutase (PGM) and phosphomannomutase activity. This activity is absent in 1291d and 1291e but is restored upon introduction of pSG30. PGM encoded by pSG34, a subclone of pSG30, was able to complement Escherichia coli PGM1, a strain deficient in PGM, as determined by bacteriophage C21 plaque formation. A revertant of 1291d that binds monoclonal antibody 2-1-L8 (specific for a 3.6-kDa LOS component) was isolated. The construction of a site-specific deletion of this region in the chromosome of 1291 confirms the role of this open reading frame in LOS biosynthesis.
Sandlin
R C
RC
Department of Microbiology, University of Maryland, College Park 20742.
Stein
D C
DC
D000595:Amino Acid Sequence; D001483:Base Sequence; D005816:Genetic Complementation Test; D008070:Lipopolysaccharides / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D016366:Open Reading Frames / Q000235:genetics; D010733:Phosphoglucomutase / Q000235:genetics / Q000378:metabolism*; D017875:Phosphotransferases (Phosphomutases) / Q000235:genetics / Q000378:metabolism*; D011709:Pyocins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid
D008070:Lipopolysaccharides; D009844:Oligosaccharides; D011709:Pyocins; D017875:Phosphotransferases (Phosphomutases); D010733:Phosphoglucomutase; C052799:phosphomannomutase