title: Loss of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferase  3 and reduced O-glycosylation in colon carcinoma cells selected  for hepatic metastasis.
abstract: O-glycosylation of mucin is initiated by the attachment of N-acetyl-D-galactosamine (GalNAc) to serine or threonine residues in mucin core polypeptides by UDPGalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGalNAc-Ts). It is not well understood how GalNAc attachment is regulated by multiple ppGalNAc-Ts in each cell. In the present study, the expression levels of murine ppGalNAc-Ts (mGalNAc-Ts), T1, T2, T3, T4, T6, and T7 were compared between mouse colon carcinoma colon 38 cells and variant SL4 cells, selected for their metastatic potentials, by using the competitive RT-PCR method. The expression levels of mGalNAc-T1, T2, and T7 were slightly higher in the SL4 cells than in the colon 38 cells, whereas the expression level of mGalNAc-T3 in the SL4 cells was 1.5% of that in the colon 38 cells. Products of enzymatic incorporations of GalNAc residues into FITCPTTTPITTTTK peptide by the use of microsome fractions of these cells as the enzyme source were separated and characterized for the number of attached GalNAc residues and their positions. The maximum number of attached GalNAc residues was 6 and 4 when the microsome fractions of the colon 38 cells and SL4 cells were used, respectively. When the microsome fractions of the colon 38 cells were treated with a polyclonal antibody raised against mGalNAc-T3, the maximum number of incorporated GalNAc residues was 4. These results strongly suggest that mGalNAc-T3 in colon 38 cells is involved in additional transfer of GalNAc residues to this peptide.
authors.affiliation: Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Kato
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Takeuchi
authors.forename: Akira
authors.initials: A
authors.lastname: Kanoh
authors.forename: Naoki
authors.initials: N
authors.lastname: Miyahara
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nemoto-Sasaki
authors.forename: Megumi
authors.initials: M
authors.lastname: Morimoto-Tomita
authors.forename: Azusa
authors.initials: A
authors.lastname: Matsubara
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Ohashi
authors.forename: Michihiko
authors.initials: M
authors.lastname: Waki
authors.forename: Katsuaki
authors.initials: K
authors.lastname: Usami
authors.forename: Ulla
authors.initials: U
authors.lastname: Mandel
authors.forename: Henrik
authors.initials: H
authors.lastname: Clausen
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Higashi
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Irimura
mesh_terms: D000116:Acetylgalactosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008113:Liver Neoplasms / Q000556:secondary*; D051379:Mice; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology; D008969:Molecular Sequence Data; D009077:Mucins / Q000378:metabolism; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D000116:Acetylgalactosamine; D000906:Antibodies; D006020:Glycopeptides; D037102:Lectins; D009077:Mucins; D017350:N-Acetylgalactosaminyltransferases; D012333:RNA, Messenger; D000097763:Polypeptide N-acetylgalactosaminyltransferase