The role of nuclear factor NF-Y/CP1 in the transcriptional regulation of the human aldehyde dehydrogenase 2-encoding gene

Mark J. Stewart, Katrina M. Dipple, Trent R. Stewart, David Crabb

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Mitochondrial aldehyde dehydrogenase (ALDH2) activity is produced at low levels in many tissues, with highest production in liver. Transfection assays using the first 600 bp of upstream DNA provided evidence for both positive and negative regulatory elements in the proximal promoter. A region from -79 to -116 bp was protected in DNase I footprinting assays and bound in electrophoretic mobility shift assays (EMSA) by a nuclear factor found in all cell lines and tissues tested. This region, denoted FP160, contained the consensus recognition sites for Sp1 and AP2, and a CCAAT box. The CCAAT box was specifically protected by a nuclear factor in methylation interference assays. Mutagenesis of specific bp within the CCAAT box eliminated protein binding in vitro and decreased transcriptional activity from the ALDH2 promoter approximately 50% in reporter gene assays. Competition experiments showed that the nuclear factor binding to the FP160 oligodeoxyribonucleotide (oligo) was competed by oligos corresponding to an NF-Y/CP1-binding site to a greater extent than by those containing sites for CTF/NF1, C/EBP or CP2. The heat stability, resistance to proteinase K digestion, sensitivity to inhibition of DNA binding by o-phenanthroline, and immunological properties of the liver factor binding to FP160 were very similar to the corresponding properties of NF-Y/CP1. Thus, the proximal ALDH2 promoter was bound by NF-Y/CP1 and this transcription factor may be responsible for the basal expression of the gene observed in most tissues. The NF-Y/CP1 present in rat liver has similar properties to that previously characterized in M12 B-lymphoma cells and LMTK mouse fibroblasts.

Original languageEnglish
Pages (from-to)155-161
Number of pages7
JournalGene
Volume173
Issue number2
DOIs
StatePublished - Sep 16 1996

Fingerprint

Aldehyde Dehydrogenase
Liver
Genes
Endopeptidase K
Oligodeoxyribonucleotides
Deoxyribonuclease I
DNA
B-Cell Lymphoma
Electrophoretic Mobility Shift Assay
Reporter Genes
Protein Binding
Mutagenesis
Methylation
Transfection
Digestion
Transcription Factors
Fibroblasts
Hot Temperature
Binding Sites
Gene Expression

Keywords

  • Alcohol metabolism
  • CCAAT box
  • DNA-binding proteins
  • Gene expression
  • Liver genes
  • Transcription factors

ASJC Scopus subject areas

  • Genetics

Cite this

The role of nuclear factor NF-Y/CP1 in the transcriptional regulation of the human aldehyde dehydrogenase 2-encoding gene. / Stewart, Mark J.; Dipple, Katrina M.; Stewart, Trent R.; Crabb, David.

In: Gene, Vol. 173, No. 2, 16.09.1996, p. 155-161.

Research output: Contribution to journalArticle

Stewart, Mark J. ; Dipple, Katrina M. ; Stewart, Trent R. ; Crabb, David. / The role of nuclear factor NF-Y/CP1 in the transcriptional regulation of the human aldehyde dehydrogenase 2-encoding gene. In: Gene. 1996 ; Vol. 173, No. 2. pp. 155-161.
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