Transcriptional regulation of N-acetylglucosaminyltransferase V by the src oncogene

Phillip Buckhaults, Lin Chen, Nevis L. Fregien, Michael Pierce

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

Transformation of baby hamster kidney fibroblasts by the Rous sarcoma virus causes a significant increase in the GlcNAcβ(1,6)Man-branched oligosaccharides by elevating the activity and mRNA transcript levels encoding N-acetylglucosaminyltransferase V (GlcNAc-T V). Elevated activity and mRNA levels could be inhibited by blocking cell proliferation with herbimycin A, demonstrating that Src kinase activity can regulate GlcNAc-T V expression. 5' RACE analysis was used to identify a 3-kilobase 5'- untranslated region from GlcNAc-T V mRNA and locate a transcriptional start site in a 25-kilobase pair GlcNAc-T V human genomic clone. A 6-kilobase pair fragment of the 5' region of the gene contained AP-1 and PEA3/Ets binding elements and, when co-transfected with a src expression plasmid into HepG2 cells, conferred src-stimulated transcriptional enhancement upon a luciferase reporter gene. This stimulation by src could be antagonized by co- transfection with a dominant-negative mutant of the Raf kinase, suggesting the involvement of Ets transcription factors in the regulation of GlcNAe-T V gene expression. The src-responsive element was localized by 5' deletion analysis to a 250-base pair region containing two overlapping Ets sites. src stimulation of transcription from this region was inhibited by co- transfection with a dominant-negative mutant of Ets-2, demonstrating that the effects of the src kinase on GlcNAc-T V expression are dependent on Ets.

Original languageEnglish
Pages (from-to)19575-19581
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number31
DOIs
StatePublished - Aug 1 1997

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alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase
src Genes
src-Family Kinases
Messenger RNA
Transfection
Genes
raf Kinases
Proto-Oncogene Proteins c-ets
Rous sarcoma virus
5' Untranslated Regions
Transcription Factor AP-1
Hep G2 Cells
Cell proliferation
Transcription
Fibroblasts
Luciferases
Oligosaccharides
Reporter Genes
Viruses
Gene expression

ASJC Scopus subject areas

  • Biochemistry

Cite this

Transcriptional regulation of N-acetylglucosaminyltransferase V by the src oncogene. / Buckhaults, Phillip; Chen, Lin; Fregien, Nevis L.; Pierce, Michael.

In: Journal of Biological Chemistry, Vol. 272, No. 31, 01.08.1997, p. 19575-19581.

Research output: Contribution to journalArticle

Buckhaults, Phillip ; Chen, Lin ; Fregien, Nevis L. ; Pierce, Michael. / Transcriptional regulation of N-acetylglucosaminyltransferase V by the src oncogene. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 31. pp. 19575-19581.
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