title: UDP-Gal/UDP-GlcNAc chimeric transporter complements mutation defect in mammalian cells deficient in UDP-Gal transporter.
abstract: The role of UDP-galactose transporter (UGT; SLC35A2) and UDP-N-acetylglucosamine transporter (NGT; SLC35A3) in glycosylation of macromolecules may be coupled and either of the transporters may partially replace the function played by its partner. The aim of this study was to construct chimeric transporters composed of the N-terminal portion of human NGT and the C-terminal portion of human UGT1 or UGT2 (NGT-UGT1 or NGT-UGT2, respectively), as well as of the N-terminal portion of UGT and C-terminal portion of NGT (UGT-NGT), overexpress them stably in UGT-deficient CHO-Lec8 and MDCK-RCA(r) cells, and characterize their involvement in glycosylation. Two chimeric proteins, NGT-UGT1 and NGT-UGT2, did not overexpress properly. In contrast, UGT-NGT chimeric protein was successfully overexpressed and localized properly to the Golgi apparatus. UGT-NGT chimeric transporter delivered UDP-Gal to the Golgi vesicles of UGT-deficient cells, which resulted in the increased content of galactosylated structures to such an extent that the wild-type phenotypes were completely restored. Our data further support our hypothesis that UGT and NGT cooperate in the UDP-Gal delivery for glycosyltransferases located in the Golgi apparatus. In a wider context, the results gained in this study add to our understanding of glycosylation, one of the basic posttranslational modifications, which affects the majority of macromolecules.
authors.affiliation: Laboratory of Biochemistry, Faculty of Biotechnology, University of Wrocław, 2 Tamka St, 50-137 Wrocław, Poland. Mariusz.Olczak@biotech.uni.wroc.pl
authors.forename: Mariusz
authors.initials: M
authors.lastname: Olczak
authors.forename: Dorota
authors.initials: D
authors.lastname: Maszczak-Seneczko
authors.forename: Paulina
authors.initials: P
authors.lastname: Sosicka
authors.forename: Piotr
authors.initials: P
authors.lastname: Jakimowicz
authors.forename: Teresa
authors.initials: T
authors.lastname: Olczak
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D006031:Glycosylation; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D017386:Sequence Homology, Amino Acid; D014531:Uridine Diphosphate Galactose / Q000378:metabolism*; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000031:analogs & derivatives* / Q000378:metabolism; D000099237:UDP-Galactose Translocators
chemical_list: D017931:DNA Primers; D009004:Monosaccharide Transport Proteins; D014531:Uridine Diphosphate Galactose; D014536:Uridine Diphosphate N-Acetylgalactosamine; D000099237:UDP-Galactose Translocators; C051472:uridine diphosphate galactosamine