C-terminal half of Salmonella enterica WbaP (RfbP) is the galactosyl-1-phosphate transferase domain catalyzing the first step of O-antigen synthesis.
We previously showed that the product of the wbaP gene of Salmonella enterica serovar Typhimurium has two functions: it is involved in the first step of O-antigen synthesis (the galactosyltransferase [GT] function) and in a later step (the T function), first thought to be the flipping of the O-antigen subunit on undecaprenyl pyrophosphate from the cytoplasmic face to the periplasmic face of the cytoplasmic membrane. We now locate two wbaP(T) mutations within the first half of the wbaP gene by sequencing. Both mutants retain GT activity, although one was a frameshift mutation resulting in a stop codon 10 codons after the frameshift to give an open reading frame containing only 138 of the 476 codons in WbaP. We also show that there is a secondary translation starting within the wbaP gene resulting in the synthesis of a polypeptide with GT activity. These results indicate that the N- and C-terminal halves of WbaP are the T and GT functional domains, respectively. We now propose that the T block operates prior to the flippase function, probably at the release of undecaprenyl pyrophosphate-linked galactose from WbaP.
Wang
L
L
Department of Microbiology, The University of Sydney, New South Wales, Australia.
Liu
D
D
Reeves
P R
PR
D000595:Amino Acid Sequence; D001483:Base Sequence; D002240:Carbohydrate Sequence; D018388:Codon, Terminator; D004252:DNA Mutational Analysis; D016368:Frameshift Mutation; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D019081:O Antigens / Q000096:biosynthesis*; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D017855:Transferases (Other Substituted Phosphate Groups) / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D018388:Codon, Terminator; D019081:O Antigens; D017855:Transferases (Other Substituted Phosphate Groups); C088138:undecaprenyl-phosphate galactosephosphotransferase