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J. Biol. Chem., Vol. 277, Issue 25, 22829-22838, June 21, 2002

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The Caenorhabditis elegans Gene, gly-2, Can Rescue the N-Acetylglucosaminyltransferase V Mutation of Lec4 Cells^*

Charles E. Warren, Aldis Krizus, Peter J. Roy^§¶, Joseph G. Culotti^§, and James W. Dennis^§

From the  Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario M5G 1X5 and ^§ Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario, M55 1A8, Canada

UDP-N-acetylglucosamine:-6-D-mannoside -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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF154122 (gly-2), AY037800 (cm20c4), and AY037802 (yk126h8). Appropriate data have also been contributed to ACeDB.

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