Cloning the 5' flanking region of neuron-specific Hel-N1: evidence for positive regulatory elements governing cell-specific transcription.

Academic Article

Abstract

  • A 5.4 kilobase-pair segment of DNA flanking the 5' end of Hel-N1 was isolated and characterized. Primer extension studies with normal human brain and neuroblastoma cells revealed a major and minor transcription-initiation site. Sequence analysis of the initial 536 bp upstream to the major start site revealed a core promoter (-1 to -181) which contained two CCAAT boxes, a weakly-conserved TATA box, and an SP1 site. This region was also moderately GC-rich (62%). Using a transient luciferase-reporter-gene assay, the core promoter was found to be essential for basal transcription both in neural (PC12) and non-neural (HeLa and glial) cell types. Two positive regulatory elements, however, were identified in the initial 536 bp (-1 to -181 and -182 to -350) which produced a five- to six-fold increase in transcriptional activity in PC12 cells vs. HeLa or glial cells. These elements, therefore, were sufficient to confer cell-specific enhanced transcription and likely contribute to the neuronal specificity of Hel-N1 mRNA expression.
  • Authors

    Published In

  • Brain Research  Journal
  • Keywords

  • Animals, Base Sequence, Cell Line, ELAV Proteins, ELAV-Like Protein 2, Gene Expression, Humans, Molecular Sequence Data, Nerve Tissue Proteins, Neurons, PC12 Cells, Promoter Regions, Genetic, RNA-Binding Proteins, Rats, Transcription, Genetic
  • Digital Object Identifier (doi)

    Pubmed Id

  • 16602715
  • Author List

  • King PH
  • Start Page

  • 141
  • End Page

  • 147
  • Volume

  • 723
  • Issue

  • 1-2