Characterization of the mitochondrial aconitase promoter and the identification of transcription factor binding

Prostate. 2006 Jul 1;66(10):1061-9. doi: 10.1002/pros.20442.

Abstract

Background: Mitochondrial (m) aconitase plays an important role in the unique pathway of citrate accumulation in prostate epithelial cells through its limited activity. In the current study, we characterized the human m-aconitase gene promoter.

Methods: A 1,411-bp 5'-flanking fragment of the human m-aconitase gene was cloned, followed by 5' serial deletion analysis of promoter activity. Transcriptional start sties and transcription factors bound to the promoter were identified by 5' RACE, DNA pull-down assay and transcription factor array analysis.

Results: Two transcriptional start sites were identified. The promoter fragment pulled down 15 transcription factors, some without consensus sequences in the promoter. Deletion of one Sp1 site eliminated all promoter activity.

Conclusions: The m-aconitase promoter is contained in a 153-bp 5' fragment lacking a TATA or CAAT sequence. Sp1 binding to a specific Sp1 site is required for promoter activity while other transcription factors are recruited through protein-protein interactions.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aconitate Hydratase / analysis
  • Aconitate Hydratase / genetics*
  • Aconitate Hydratase / physiology
  • Base Sequence
  • CCAAT-Binding Factor / genetics
  • CCAAT-Binding Factor / physiology
  • Cell Line, Tumor
  • DNA / analysis
  • DNA / genetics
  • Epithelium / enzymology
  • Gene Expression Regulation
  • Humans
  • Male
  • Mitochondria / enzymology*
  • Molecular Sequence Data
  • Oligonucleotide Array Sequence Analysis
  • Promoter Regions, Genetic / genetics*
  • Promoter Regions, Genetic / physiology
  • Prostate / enzymology
  • Protein Binding / genetics
  • Protein Binding / physiology
  • Sp1 Transcription Factor / analysis
  • Sp1 Transcription Factor / genetics
  • Sp1 Transcription Factor / physiology
  • TATA-Box Binding Protein / genetics
  • TATA-Box Binding Protein / physiology
  • Transcription Factors / analysis
  • Transcription Factors / genetics*
  • Transcription Factors / physiology
  • Transcription, Genetic / genetics
  • Transcription, Genetic / physiology

Substances

  • CCAAT-Binding Factor
  • Sp1 Transcription Factor
  • TATA-Box Binding Protein
  • Transcription Factors
  • DNA
  • Aconitate Hydratase