title: DNA-mediated transformation of N-acetylglucosaminyltransferase I activity into an enzyme deficient cell line.
abstract: N-acetylglucosaminyltransferase I (GlcNAc-TI) catalyzes the first reaction in the conversion of ASN-linked cell surface oligosaccharides from a mannose-terminating structure to more complex carbohydrate structures. The mutant Chinese hamster ovary (CHO) cell line, Lec1, is deficient in this enzyme and, therefore, shows increased sensitivity to the lectin, Concanavalin A, which binds to the mannose-terminating oligosaccharides that accumulate on Lec1 cell surface glycoproteins. Spontaneous revertants of the Lec1 phenotype have never been observed. We report here the isolation of stable revertants of Lec1 cells to the parental CHO cell lectin-resistance phenotype after DNA-mediated transformation with human DNA. Both primary and secondary transformants express varying levels of GlcNAc-TI enzyme activity which was stable even when the cells were cultured in nonselective conditions. Human alu repeat DNA sequences are present in the primary transformants, but these sequences could not be detected in the secondary transformants.
authors.affiliation: Department of Anatomy and Cell Biology, University of Miami School of Medicine, FL 33101.
authors.forename: J
authors.initials: J
authors.lastname: Ripka
authors.forename: M
authors.initials: M
authors.lastname: Pierce
authors.forename: N
authors.initials: N
authors.lastname: Fregien
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed / Q000201:enzymology; D003208:Concanavalin A; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA; D005260:Female; D005964:Glucosyltransferases / Q000172:deficiency* / Q000378:metabolism; D006801:Humans; D017351:N-Acetylglucosaminyltransferases; D010053:Ovary; D014162:Transfection; D014170:Transformation, Genetic; D014909:Wheat Germ Agglutinins
chemical_list: D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D004247:DNA; D005964:Glucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C024850:alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I