Functional identification of the promoter of the gene encoding the Rhesus monkey β-amyloid precursor protein

Weihong Song, Debomoy K. Lahiri

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

Misregulation of transcription of the β-amyloid precursor protein (βAPP) gene is implicated in the pathogenesis of Alzheimer's disease (AD). Here we characterize the 5'-flanking region, the first exon and intron of the βAPP gene of the Rhesus monkey (rhβAPP). For functional analysis, transient transfection in PC12 cells was performed with a series of 5'-deletion constructs (fused with a reporter gene), that extended as far upstream as -7900 down to -1 bp. Chloramphenicol acetyltransferase/promoter fusion assays showed that both -7900/+104 and -75/+104-bp regions possessed strong promoter activity. However, -2542/+104 bp had the strongest promoter activity, whereas -204/+104 bp showed a major reduction in activity and -47/+104 bp showed almost a complete loss of activity. A region from -75 to +104 bp was essential for minimal basic promoter activity because mutation at the activating site of an upstream stimulator factor (USF) within this region abolished the promoter activity. The very upstream region (-5529/-3416 bp) displayed a negative effect on promoter activity. Two blocks of the sequence, 641 bp (-1131/-490) and 105 bp (-309/-204), acted as positive regulators for promoter activity. Another 61-bp block (-204/-143) acted as a negative regulator. Gel shift assay indicated that the -249-242-bp region contains a binding domain for the AP-2 transcription factor. No second promoter or bidirectional promoter was observed. A region spanning the first exon and part of the first intron (+99 to +6800 bp) acted as a negative regulator. These results suggest that a region of -75 to +104 bp, which contains the pyrimidine-rich initiator element, the 5'-untranslated region and the binding site for USF, constitute the minimal promoter element and that interactions between multiple positive and negative elements, the USF and initiator element are crucial for transcription of the TATA-less βAPP promoter.

Original languageEnglish (US)
Pages (from-to)165-176
Number of pages12
JournalGene
Volume217
Issue number1-2
DOIs
StatePublished - Sep 14 1998

Keywords

  • APP intron
  • APP promoter
  • Primate
  • Promoter activity
  • Regulatory elements
  • Serial deletion
  • Transcription factor
  • Transcriptional regulator
  • Upstream sequence

ASJC Scopus subject areas

  • Genetics

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