title: Novel functions for glycosyltransferases Jhp0562 and GalT in Lewis antigen synthesis and variation in Helicobacter pylori.
abstract: Lewis (Le) antigens are fucosylated oligosaccharides present in the Helicobacter pylori lipopolysaccharide. Expression of these antigens is believed to be important for H. pylori colonization, since Le antigens also are expressed on the gastric epithelia in humans. A galactosyltransferase encoded by β-(1,3)galT is essential for production of type 1 (Le(a) and Le(b)) antigens. The upstream gene jhp0562, which is present in many but not all H. pylori strains, is homologous to β-(1,3)galT but is of unknown function. Because H. pylori demonstrates extensive intragenomic recombination, we hypothesized that these two genes could undergo DNA rearrangement. A PCR screen and subsequent sequence analyses revealed that the two genes can recombine at both the 5' and 3' ends. Chimeric β-(1,3)galT-like alleles can restore function in a β-(1,3)galT null mutant, but neither native nor recombinant jhp0562 can. Mutagenesis of jhp0562 revealed that it is essential for synthesis of both type 1 and type 2 Le antigens. Transcriptional analyses of both loci showed β-(1,3)galT expression in all wild-type (WT) and mutant strains tested, whereas jhp0562 was not expressed in jhp0562 null mutants, as expected. Since jhp0562 unexpectedly displayed functions in both type 1 and type 2 Le synthesis, we asked whether galT, part of the type 2 synthesis pathway, had analogous functions in type 1 synthesis. Mutagenesis and complementation analysis confirmed that galT is essential for Le(b) production. In total, these results demonstrate that galT and jhp0562 have functions that cross the expected Le synthesis pathways and that jhp0562 provides a substrate for intragenomic recombination to generate diverse Le synthesis enzymes.
authors.affiliation: maryann.pohl@einstein.yu.edu
authors.forename: Mary Ann
authors.initials: MA
authors.lastname: Pohl
authors.forename: Sabine
authors.initials: S
authors.lastname: Kienesberger
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Blaser
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D005816:Genetic Complementation Test; D014644:Genetic Variation; D005838:Genotype; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D016480:Helicobacter pylori / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D007983:Lewis Blood Group Antigens / Q000096:biosynthesis / Q000276:immunology; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011995:Recombination, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D007983:Lewis Blood Group Antigens; D008070:Lipopolysaccharides; C442865:lipopolysaccharide, Helicobacter pylori; D016695:Glycosyltransferases