DNA-mediated transformation of N-acetylglucosaminyltransferase I activity into an enzyme deficient cell line

James Ripka, Michael Pierce, Nevis L. Fregien

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)554-560
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume159
Issue number2
DOIs
StatePublished - Mar 15 1989
Externally publishedYes

Fingerprint

Cells
Mannose
Cricetulus
Oligosaccharides
Lectins
Cell Line
Ovary
DNA
Enzymes
Conversion Disorder
Phenotype
DNA sequences
Membrane Glycoproteins
Enzyme activity
Concanavalin A
Cultured Cells
Carbohydrates
alpha-1,3-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase I

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

DNA-mediated transformation of N-acetylglucosaminyltransferase I activity into an enzyme deficient cell line. / Ripka, James; Pierce, Michael; Fregien, Nevis L.

In: Biochemical and Biophysical Research Communications, Vol. 159, No. 2, 15.03.1989, p. 554-560.

Research output: Contribution to journalArticle

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