Characterization of the serogroup O11 O-antigen locus of Pseudomonas aeruginosa PA103.
We previously cloned a genomic DNA fragment from the serogroup O11 Pseudomonas aeruginosa strain PA103 that contained all genes necessary for O-antigen synthesis and directed the expression of serogroup O11 antigen on recombinant Escherichia coli and Salmonella. To elucidate the pathway of serogroup O11 antigen synthesis, the nucleotide sequence of the biosynthetic genes was determined. Eleven open reading frames likely to be involved in serogroup O11 O-antigen biosynthesis were identified and are designated in order as wzzPaO111 (wzz from P. aeruginosa serogroup O11), wzxPaO11, wbjA, wzyPaO11, wbjB to wbjF, wbpLO11 and wbpMO11 (wbpL and wbpM from serogroup O11). Consistent with previous descriptions of O-antigen biosynthetic gene loci, the entire region with the exception of wbpMO11 has a markedly reduced G+C content relative to the chromosomal average. WzyPaO11 shows no significant similarity at the protein or DNA sequence level to any database sequence and is very hydrophobic, with 10 to 12 putative transmembrane domains, both typical characteristics of O-antigen polymerases. A nonpolar chromosomal insertion mutation in wzyPaO11 in P. aeruginosa PA103 confirmed the identity of this gene. There is striking similarity between WbjBCDE and Cap(5/8)EFGL, involved in type 5 and type 8 capsule biosynthesis in Staphylococcus aureus. There is nearly total identity between wbpMO11 and wbpMO5, previously shown by others to be present in all 20 P. aeruginosa serogroups. Using similarity searches, we have assigned functions to the proteins encoded by the PA103 O-antigen locus and present the potential steps in the pathway for the biosynthesis of P. aeruginosa serogroup O11 O antigen.
Departments of Microbiology, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908, USA.
C R
CR
Dean
C V
CV
Franklund
J D
JD
Retief
M J
MJ
Coyne
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K
Hatano
D J
DJ
Evans
G B
GB
Pier
J B
JB
Goldberg
D001426:Bacterial Proteins / Q000235:genetics; D015153:Blotting, Western; D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D019081:O Antigens / Q000096:biosynthesis*; D016366:Open Reading Frames; D011550:Pseudomonas aeruginosa / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012703:Serotyping
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D019081:O Antigens; C095976:O antigen, Pseudomonas