UDP-N-acetylglucosamine:alpha-3-D-mannoside
beta-1,2-N-acetylglucosaminyltransferase I (GnT I) and UDP-N-acetylglucosamine:alpha-6-D-mannoside
beta-1,2-N-acetylglucosaminyltransferase II (GnT 11) are key enzymes in the synthesis of Asn-linked hybrid and complex glycans. We have cloned cDNAs from Caenorhabditis elegans for three genes homologous to mammalian GnT I (designated
gly-12,
gly-13 and
gly-14) and one gene homologous to mammalian GnT II. All four cDNAs encode proteins which have the domain structure typical of previously cloned Golgi-type glycosyltransferases and show enzymatic activity (GnT I and GnT 11, respectively) on expression in transgenic worms. We have isolated worm mutants lacking the three GnT I genes by the method of ultraviolet irradiation in the presence of trimethylpsoralen (TMP); null mutants for GnT 11 have not yet been obtained. The
gly-12 and
gly-14 mutants as well as the
gly-14;
gly-12 double mutant displayed wild-type phenotypes indicating that neither
gly-12 nor
gly-14 is necessary for worm development under standard laboratory conditions. This finding and other data indicate that the GLY-13 protein is the major functional GnT I in C elegans. The mutation lacking the
gly-13 gene is partially lethal and the few survivors display severe morphological and behavioral defects. We have shown that the observed phenotype co-segregates with the
gly-13 deletion in genetic mapping experiments although a second mutation near the
gly-13 gene cannot as yet be ruled out. Our data indicate that complex and hybrid N-glycans may play critical roles in the morphogenesis of C elegans, as they have been shown