An Arabidopsis thaliana cDNA complements the N-acetylglucosaminyltransferase I deficiency of CHO Lec1 cells.
N-Acetylglucosaminyltransferase I (GlcNAcT-I, EC 2.4.1.101) is the enzyme which initiates the formation of complex N-linked glycans in eukaryotes by transforming GlcNAc to the oligo-mannosyl acceptor Man(5)GlcNAc(2)-Asn. The enzymatic activity and the structure that is synthesised by this enzyme are found in animals and plants but not in yeast. cDNAs encoding the enzyme have already been cloned from several mammals and the nematode Caenorhabditis elegans. In this article the cloning of an Arabidopsis thaliana GlcNAcT-I cDNA with homology to animal cDNAs is described. By expression of the plant cDNA in CHO Lec1 cells, a mammalian cell line deficient in GlcNAcT-I, it was shown that it encodes an active enzyme with the same enzymatic activity as the animal homologue. It has already been shown that a human GlcNAcT-I can complement an A. thaliana mutant (cgl-1). Here it is shown that the reverse is also true, the plant glycosyltransferase is able to complement a mammalian mutant (Lec1) deficient in GlcNAcT-I.
Bakker
H
H
Department of Molecular Biology, Centre for Plant Breeding and Reproduction Research (CPRO-DLO), Wageningen, 6700AA, The Netherlands. h.bakker@cpro.dlo.nl
Lommen
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A
Jordi
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Stiekema
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Bosch
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D
D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D016466:CHO Cells / Q000201:enzymology*; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000737:chemistry / Q000172:deficiency / Q000235:genetics*; D016415:Sequence Alignment; D014162:Transfection
D018076:DNA, Complementary; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I