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T-cell-specific deletion of a polypeptide N-acetylgalactosaminyl-transferase gene by site-directed recombination.

UDP-N-acetylgalactosamine (GalNAc): polypeptide N-acetylgalactosaminyltransferase (polypeptide GalNAc-T) catalyzes transfer of the monosaccharide GalNAc to serine and threonine residues, thereby initiating O-linked oligosaccharide biosynthesis. Previous studies have suggested the possibility of multiple polypeptide GalNAc-Ts, although attachment of saccharide units to polypeptide or lipid in generating oligosaccharide structures in vertebrates has been dependent upon the activity of single gene products. To address this issue and to determine the relevance of Oglycosylation variation in T-cell ontogeny, we have directed Cre/loxP mutagenic recombination to the polypeptide GalNAc-T locus in gene-targeted mice. Resulting deletion in the catalytic region of polypeptide GalNAc-T occurred to completion on both alleles in thymocytes and was found in peripheral T cells, but not among other cell types. Thymocyte O-linked oligosaccharide formation persisted in the absence of a functional targeted polypeptide GalNAc-T allele as determined by O-glycan-specific lectin binding. T-cell development and colonization of secondary lymphoid organs were also normal. These results indicate a complexity in vertebrate O-glycan biosynthesis that involves multiple polypeptide GalNAc-Ts. We infer the potential for protein-specific O-glycan formation governed by distinct polypeptide GalNAc-Ts.

Pubmed ID: 8618846


  • Hennet T
  • Hagen FK
  • Tabak LA
  • Marth JD


Proceedings of the National Academy of Sciences of the United States of America

Publication Data

December 19, 1995

Associated Grants

  • Agency: NIDCR NIH HHS, Id: DE08108

Mesh Terms

  • Alleles
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cattle
  • Cloning, Molecular
  • Consensus Sequence
  • Glycosylation
  • Introns
  • Mice
  • Mice, Transgenic
  • Molecular Sequence Data
  • N-Acetylgalactosaminyltransferases
  • Oligopeptides
  • Oligosaccharides
  • Recombinant Fusion Proteins
  • Recombination, Genetic
  • Restriction Mapping
  • Sequence Deletion
  • Spleen
  • Substrate Specificity
  • T-Lymphocytes
  • Vertebrates