Functional analysis of promoter activity of murine β-1,6-N-acetylglucosaminyltransferase

Guoyun Chen, Nobuyuki Kurosawa, Takashi Muramatsu

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

In the mouse, β-1,6-N-acetylglucosaminyltransferase (IGnT) forms branches in poly-N-acetyllactosamines, which are good scaffolds for oncodevelopmental cell-surface antigens and recognition markers. There are two isoforms of IGnT, IGnT A and B, which are produced by alternative splicing of the IGnT gene; the unique portion is encoded by exon 1 and common portion is encoded by exons 2 and 3. Thus, the expression of each isoform is controlled by a different promoter. Here, we identified the regulatory regions of the mouse IGnT A and B genes. The promoter regions for IGnT A and B did not contain putative TATA or CAAT boxes, but each contained GT boxes. The upstream regulatory region of each gene was examined by transient luciferase reporter gene transfection experiments and gel mobility shift assay. Promoter activity for each gene was detected in F9 embryonal carcinoma cells, which express IGnT A and B, but not in N2a cells, which do not express the gene. Deletion analysis demonstrated that the regions 308 bp upstream from the transcriptional initiation site of IGnT A and 430 bp upstream from the transcriptional initiation site of IGnT B showed minimal promoter activity. Mutation of the single GT box in IGnT A and two GT boxes in IGnT B caused marked reduction of the promoter activity. These findings provided strong evidence that the GT boxes play crucial roles in transcriptional regulation of the genes.

Original languageEnglish (US)
Pages (from-to)253-259
Number of pages7
JournalGene
Volume275
Issue number2
DOIs
StatePublished - Sep 19 2001

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Genes
Nucleic Acid Regulatory Sequences
Exons
Protein Isoforms
Embryonal Carcinoma Stem Cells
Alternative Splicing
Electrophoretic Mobility Shift Assay
Surface Antigens
Luciferases
Reporter Genes
Genetic Promoter Regions
Transfection
N-acetyllactosaminide beta-1,6-N-acetylglucosaminyltransferase
Gels
Mutation
poly-N-acetyllactosamine
N-acetylglucosaminyltransferase IGnT

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Functional analysis of promoter activity of murine β-1,6-N-acetylglucosaminyltransferase. / Chen, Guoyun; Kurosawa, Nobuyuki; Muramatsu, Takashi.

In: Gene, Vol. 275, No. 2, 19.09.2001, p. 253-259.

Research output: Contribution to journalArticle

Chen, Guoyun ; Kurosawa, Nobuyuki ; Muramatsu, Takashi. / Functional analysis of promoter activity of murine β-1,6-N-acetylglucosaminyltransferase. In: Gene. 2001 ; Vol. 275, No. 2. pp. 253-259.
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