title: Cloning and expression of a brain-specific putative UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferase gene.
abstract: We isolated a rat cDNA clone and its human orthologue, which are most homologous to UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferase 9, by homology-based PCR from brain. Nucleotide sequence analysis of these putative GalNAc-transferases (designated pt-GalNAc-T) showed that they contained structural features characteristic of the GalNAc-transferase family. It was also found that human pt-GalNAc-T was identical to the gene WBSCR17, which is reported to be in the critical region of patients with Williams-Beuren Syndrome, a neurodevelopmental disorder, and to be predominantly expressed in brain and heart. In order to investigate the expression of pt-GalNAc-T in brain in more detail, we first examined that of human pt-GalNAc-T by Northern blot analysis and found the expression of the 5.0-kb mRNA to be most abundant in cerebral cortex with somewhat less abundant in cellebellum. The expression of rat pt-GalNAc-T was investigated more extensively. The brain-specific expression of 2.0-kb and 5.0-kb transcripts was demonstrated by Northern blot analysis. In situ hybridization in the adult brain revealed high levels of expression in cerebellum, hippocampus, thalamus, and cerebral cortex. Moreover, observation at high magnification revealed the expression to be associated with neurons, but not with glial cells. Analysis of the rat embryos also demonstrated that rat pt-GalNAc-T was expressed in the nervous system, including in the diencephalons, cerebellar primordium, and dorsal root ganglion. However, recombinant human pt-GalNAc-T, which was expressed in insect cells, did not glycosylate several peptides derived from mammalian mucins, suggesting that it may have a strict substrate specificity. The brain-specific expression of pt-GalNAc-T suggested its involvement in brain development, through O-glycosylation of proteins in the neurons.
authors.affiliation: Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Kamigamo-motoyama, Kyoto, Japan.
authors.forename: Naosuke
authors.initials: N
authors.lastname: Nakamura
authors.forename: Shinya
authors.initials: S
authors.lastname: Toba
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Hirai
authors.forename: Shinichi
authors.initials: S
authors.lastname: Morishita
authors.forename: Tadahisa
authors.initials: T
authors.lastname: Mikami
authors.forename: Morichika
authors.initials: M
authors.lastname: Konishi
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Itoh
authors.forename: Akira
authors.initials: A
authors.lastname: Kurosaka
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular / Q000379:methods*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D017350:N-Acetylgalactosaminyltransferases / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D014536:Uridine Diphosphate N-Acetylgalactosamine / Q000737:chemistry*; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D017350:N-Acetylgalactosaminyltransferases; D014536:Uridine Diphosphate N-Acetylgalactosamine; D000097763:Polypeptide N-acetylgalactosaminyltransferase