cDNA cloning and expression of a family of UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase sequence homologs from Caenorhabditis elegans. 1998

F K Hagen, and K Nehrke
Center for Oral Biology, School of Medicine and Dentistry, University of Rochester, Rochester, New York 14642, USA. fred_hagen@urmc.rochester.edu

The initiation of mucin-type O-glycosylation is catalyzed by a family of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGaNTase) (EC 2.4.1.41). By screening two mixed-stage Caenorhabditis elegans cDNA libraries, a total of 11 distinct sequence homologs of the ppGaNTase gene family were cloned, sequenced, and expressed as truncated recombinant proteins (gly-3, gly-4, gly-5a, gly-5b, gly-5c, gly-6a, gly-6b, gly-6c, gly-7, gly-8, and gly-9). All clones encoded type II membrane proteins that shared 60-80% amino acid sequence similarity with the catalytic domain of mammalian ppGaNTase enzymes. Two sets of cDNA clones (gly-5 and gly-6) contained variants that appeared to be produced by alternative message processing. gly-6c contained a reading frameshift and premature termination codon in the C-terminal lectin-like domain found in most other ppGaNTase proteins, and a second clone (gly-8) lacked the typical C-terminal region completely. Homogenates of nematodes and immunopurified preparations of the recombinant GLY proteins demonstrated that worms express functional ppGaNTase enzymes (GLY-3, GLY-4, GLY-5A, GLY-5B, and GLY-5C), which can O-glycosylate mammalian apomucin peptide sequences in vitro. In addition to demonstrating the existence of ppGaNTase enzymes in a nematode organism, the substantial diversity of these isoforms in C. elegans suggests that mucin O-glycosylation is catalyzed by a complex gene family, which is conserved among evolutionary-distinct organisms.

UI MeSH Term Description Entries
D008969 Molecular Sequence Data Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories. Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D000097763 Polypeptide N-acetylgalactosaminyltransferase Family of enzymes that catalyze the formation of GalNAcAlpha1-serine/threonine linkages in glycoproteins. Galactosylgalactosylglucosylceramide beta-D-acetylgalactosaminyltransferase,Globoside Synthase,Globoside beta GalNAc Transferase,Protein-UDPacetylgalactosaminyltransferase,(1-3)-N-acetyl-beta-galactosaminyltransferase,(1-4)-N-acetyl-beta-D-galactosaminyltransferase,4-GalNActransferase,GalNAc-T1,GalNAc-T10,GalNAc-T2,GalNAc-T3,GalNAc-T4,GalNAc-T5,GalNAc-T8,GalNAc-transferase,GalNAcT-1,GalNAcT-2,GalNAcT-4,GalNAcT-8,UDP-GPAGAT,UDP-GalNAc-beta-galactose beta 1,4-N-acetylgalactosaminyltransferase,UDP-GalNAc-polypeptide N-acetylgalactosaminyltransferase,UDP-N-acetyl-D-galactosamine polypeptide N-acetylgalactosaminyltransferase-T4,UDP-N-acetylgalactosamine mucin transferase,UDP-N-acetylgalactosamine-beta-galactose beta 1,4-N-acetylgalactosaminyltransferase,UDP-N-acetylgalactosamine-globoside beta-N-acetylgalactosaminyltransferase,UDP-N-acetylgalactosamine-globosidetriaosylceramide beta-3-N-acetylgalactosaminyltransferase,UDP-N-acetylgalactosamine-polypeptide N-acetylgalactosamine transferase,UDPacetylgalactosamine-galactosyl-galactosyl-glucosylceramide beta-N-acetyl-D-galactosaminyltransferase,UDPacetylgalactosamine-protein acetylgalactosaminyltransferase,beta-1,4-N-acetylgalactosaminyltransferase,beta-N-acetylgalactosaminyltransferase,beta1,6N-acetylgalactosaminyltransferase,polypeptide N-acetylgalactosaminyltransferase 1,polypeptide N-acetylgalactosaminyltransferase 10,polypeptide N-acetylgalactosaminyltransferase 2,polypeptide N-acetylgalactosaminyltransferase 3,polypeptide N-acetylgalactosaminyltransferase 4,polypeptide N-acetylgalactosaminyltransferase 5,polypeptide N-acetylgalactosaminyltransferase 8,pp-GalNAc-T10,ppGalNAc-T,Synthase, Globoside
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D001483 Base Sequence The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence. DNA Sequence,Nucleotide Sequence,RNA Sequence,DNA Sequences,Base Sequences,Nucleotide Sequences,RNA Sequences,Sequence, Base,Sequence, DNA,Sequence, Nucleotide,Sequence, RNA,Sequences, Base,Sequences, DNA,Sequences, Nucleotide,Sequences, RNA
D016415 Sequence Alignment The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms. Sequence Homology Determination,Determination, Sequence Homology,Alignment, Sequence,Alignments, Sequence,Determinations, Sequence Homology,Sequence Alignments,Sequence Homology Determinations
D017173 Caenorhabditis elegans A species of nematode that is widely used in biological, biochemical, and genetic studies. Caenorhabditis elegan,elegan, Caenorhabditis
D017350 N-Acetylgalactosaminyltransferases Enzymes that catalyze the transfer of N-acetylgalactosamine from a nucleoside diphosphate N-acetylgalactosamine to an acceptor molecule which is frequently another carbohydrate. EC 2.4.1.-. N-Acetylgalactosamine Transferases,N Acetylgalactosamine Transferases,N Acetylgalactosaminyltransferases,Transferases, N-Acetylgalactosamine
D017386 Sequence Homology, Amino Acid The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species. Homologous Sequences, Amino Acid,Amino Acid Sequence Homology,Homologs, Amino Acid Sequence,Homologs, Protein Sequence,Homology, Protein Sequence,Protein Sequence Homologs,Protein Sequence Homology,Sequence Homology, Protein,Homolog, Protein Sequence,Homologies, Protein Sequence,Protein Sequence Homolog,Protein Sequence Homologies,Sequence Homolog, Protein,Sequence Homologies, Protein,Sequence Homologs, Protein

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