Genomic organization and expression of the mouse equilibrative, nitrobenzylthioinosine-sensitive nucleoside transporter 1 (ENT1) gene

Doo Sup Choi, Masahisa Handa, Hannah Young, Adrienne S. Gordon, Ivan Diamond, Robert O. Messing

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We have cloned and characterized the genomic structure of the mouse gene for the NBMPR-sensitive equilibrative nucleoside transporter (mENT1), which is located on chromosome 17C. About 12-kb of genomic DNA was sequenced including the promoter region, 12 exons, 11 introns, and the 3'-untranslated region. All exon-intron junction sequences conform to the GT/AG rule. Primer extension analysis demonstrated a transcription initiation site located 252 bp upstream of the translation start site. Analysis of the 2.5-kb 5'-flanking sequence shows putative binding sites for several transcription factors, including GATA-1, IRF-2, Pit-1, myogenin, CREB, Sp-1, Ap-2, MAZ, and GR. We demonstrated that mouse ENT1 mRNA was highly expressed in heart, spleen, lung, liver, and testis. Lower levels of expression were detected in brain and kidney. Functional analysis of the 5'-flanking region showed that the nucleotide sequence from -652 to -111 contains cis-regulatory elements that promote gene expression. We found two Sp-1 binding sites (-296/-303, -307/-313) and two MAZ binding sites (-353/-359, -522/-528) in this region. Luciferase assay results suggest that MAZ and Sp-1 transcription factors are important positive regulators of transcription for the ENT1 gene in NG108-15 cells. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)200-208
Number of pages9
JournalBiochemical and Biophysical Research Communications
Volume277
Issue number1
DOIs
StatePublished - Oct 14 2000
Externally publishedYes

Fingerprint

Nucleoside Transport Proteins
Genes
5' Flanking Region
Binding Sites
Introns
Exons
Sp Transcription Factors
Equilibrative Nucleoside Transporter 1
Transcription Factors
GATA Transcription Factors
Myogenin
Functional analysis
Transcription Initiation Site
3' Untranslated Regions
Transcription
Chromosomes
Luciferases
Genetic Promoter Regions
Gene expression
Liver

Keywords

  • Chromosomal localization
  • Gene structure
  • Luciferase assay
  • MAZ
  • mRNA expression
  • Northern blot
  • Nucleoside transporter

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Genomic organization and expression of the mouse equilibrative, nitrobenzylthioinosine-sensitive nucleoside transporter 1 (ENT1) gene. / Choi, Doo Sup; Handa, Masahisa; Young, Hannah; Gordon, Adrienne S.; Diamond, Ivan; Messing, Robert O.

In: Biochemical and Biophysical Research Communications, Vol. 277, No. 1, 14.10.2000, p. 200-208.

Research output: Contribution to journalArticle

Choi, Doo Sup ; Handa, Masahisa ; Young, Hannah ; Gordon, Adrienne S. ; Diamond, Ivan ; Messing, Robert O. / Genomic organization and expression of the mouse equilibrative, nitrobenzylthioinosine-sensitive nucleoside transporter 1 (ENT1) gene. In: Biochemical and Biophysical Research Communications. 2000 ; Vol. 277, No. 1. pp. 200-208.
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