http://hdl.handle.net/1765/3035

Molecular and functional analysis of the XPBC/ERCC-3 promoter: Transcription activity is dependent on the integrity of an Sp1 binding element.


Article
volume 20 pp 217-224.
Related Files
asset icon
(eur_hoeijmakers_9050.pdf, 1.9MB)

The human XPBC/ERCC-3 gene, which corrects the excision-repair defect in xeroderma pigmentosum group B cells and the UV-sensitive CHO mutant 27-1 cells, appears to be expressed constitutively in various cell types and tissues. We have analysed the structure and functionality of the XPBC/ERCC-3 promoter. Transcription of the XPBC/ERCC-3 gene is initiated from heterogeneous sites, with a major startpoint mapped at position -54 (relative to the translation start codon ATG). The promoter region does not possess classical TATA and CAAT elements, but it is GC-rich and contains three putative Sp1-binding sites. In addition, there are two elements related to the cyclic AMP (cAMP)-response element (CRE) and the 12-O-tetradecanoyl phorbol-13-acetate-response element (TRE) in the 5'-flanking region. Transient expression analysis of XPBC/ERCC-3 promoter-CAT chimeric plasmids revealed that a 127-bp fragment, spanning position -129 to -3, is minimally required for the promoter activity. Transcription of the XPBC/ERCC-3 promoter depends on the integrity of a putative Sp1-binding site in close proximity to the major cap site. Band shift assays showed that this putative Sp1-binding site can interact specifically with a nuclear factor, most likely transcription factor Sp1 (or an Sp1-like factor) in vitro.



Keywords


Automatically Extracted Terms
  • /ercc
  • promoter
  • xpbc /ercc promoter
  • xpbc /ercc gene
  • element
  • transcription
  • sequence
  • plasmid
  • fragment
  • cap site
  • region
  • activity
  • transcription initiation site
  • factor
  • analysis
  • position
  • oligonucleotide
  • probe
  • initiation
  • assay


Bookmark with: