Immediate upstream promoter regions required for neurospecific expression of SNAP-25

Andrey E. Ryabinin, Tom N. Sato, Peter J. Morris, David S. Latchman, Michael C. Wilson

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

The promoter structure and regulation of Snap, a gene encoding the presynaptic t-SNARE SNAP-25 implicated in synaptic vesicle docking and fusion, was studied. Transcription start-site analysis revealed two major start sites located 42 nucleotides apart. Nucleotide sequence of a promoter region 2073 nucleotides upstream of the first transcription site contains three AP-1, one CRE sequence, and three Sp1-like sites close to the TATA box. Further upstream of these sites two TG repeats were found. The ability of regions within the 5′ upstream sequence to promote basal neural-specific expression in tissue culture cells was evaluated using a series of constructs containing both Snap gene start sites with progressively restricted 5′ sequence linked to the chloramphenicol acetyl transferase (CAT) reporter gene. CAT expression was maximal in neuron-like undifferentiated ND7 and PC12 cells transfected with constructs containing Snap sequences up to 127 bp from the start site. In contrast, nonneuronal fibroblast cell lines did not express significant amounts of CAT, suggesting that this short 127-bp sequence is sufficient to drive neural specific expression of SNAP-25. Band shift analysis of oligonucleotides spanning from -127 to -41 bp of the Snap promoter revealed three distinct DNA-protein complexes generated by brain nuclear extracts and one by liver nuclear extracts, indicating that transcription factors that bind to this 86-bp sequence located just upstream of the TATA box are involved in regulation of basal neurospecific expression of this gene.

Original languageEnglish (US)
Pages (from-to)201-210
Number of pages10
JournalJournal of Molecular Neuroscience
Volume6
Issue number3
DOIs
StatePublished - Sep 1 1995

Keywords

  • ND7 cell
  • PC12 cell
  • SNAP-25
  • Sp1-like sites
  • gene regulation
  • presynaptic protein
  • promoter
  • t-SNARE

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience

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