The Caenorhabditis elegans gene, gly-2, can rescue the N-acetylglucosaminyltransferase V mutation of Lec4 cells

J Biol Chem. 2002 Jun 21;277(25):22829-38. doi: 10.1074/jbc.M201390200. Epub 2002 Apr 5.

Abstract

UDP-N-acetylglucosamine:alpha-6-d-mannoside beta-1,6-N-acetylglucosaminyltransferase V (GlcNAc-TV) is a regulator of polylactosamine-containing N-glycans and is causally involved in T cell regulation and tumor metastasis. The Caenorhabditis elegans genome contains a single orthologous gene, gly-2, that is transcribed and encodes a 669-residue type II membrane protein that is 36.7% identical to mammalian GlcNAc-TV (Mgat-5). Recombinant GLY-2 possessed GlcNAc-TV activity when assayed in vitro, and protein truncations demonstrated that the N-terminal boundary of the catalytic domain is Ile-138. gly-2 complemented the Phaseolus vulgaris leucoagglutinin binding defect of Chinese hamster ovary Lec4 cells, whereas GLY-2(L116R), an equivalent mutation to that which causes the Lec4A phenotype, could not. We conclude that the worm gene is functionally interchangeable with the mammalian form. GlcNAc-TV activity was detected in wild-type animals but not those homozygous for a deletion allele of gly-2. Activity was restored in mutant animals by an extrachromosomal array that encompassed the gly-2 gene. Green fluorescent protein reporter transgenes driven by the gly-2 promoter were expressed by developing embryos from the late comma stage onward, present in a complex subset of neurons in larvae and, in addition, the spermathecal and pharyngeal-intestinal valves and certain vulval cells of adults. However, no overt phenotypes were observed in animals homozygous for deletion alleles of gly-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • CHO Cells
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins / biosynthesis
  • Caenorhabditis elegans Proteins / genetics*
  • Chromosome Mapping
  • Cricetinae
  • DNA, Complementary / metabolism
  • Gene Deletion
  • Genes, Reporter
  • Genetic Complementation Test
  • Genome, Fungal
  • Green Fluorescent Proteins
  • Homozygote
  • Humans
  • Luminescent Proteins / metabolism
  • Microscopy, Fluorescence
  • Microsomes / metabolism
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Mutation*
  • N-Acetylglucosaminyltransferases / metabolism*
  • Phenotype
  • Poly A / metabolism
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Transfection
  • Transgenes

Substances

  • Caenorhabditis elegans Proteins
  • DNA, Complementary
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • gly-2 protein, C elegans
  • Green Fluorescent Proteins
  • Poly A
  • N-Acetylglucosaminyltransferases
  • alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase

Associated data

  • GENBANK/AF154122
  • GENBANK/AY037800
  • GENBANK/AY037802