Transcriptional activation of the human rod cGMP-phosphodiesterase beta-subunit gene is mediated by an upstream AP-1 element

Nucleic Acids Res. 1997 Oct 1;25(19):3863-7. doi: 10.1093/nar/25.19.3863.

Abstract

During photoactivation retinal cGMP-phosphodiesterase (PDE) mediates signal transduction in the photoreceptor outer segments. Mutations in the beta-subunit gene of rod-specific PDE (beta-PDE) have been associated with inherited retinal degeneration in a number of species, including human. Here we have investigated the proximal upstream sequences that participate in transcriptional activation of this gene. Transient transfections demonstrated that the sequence from -72 to +53 bp contained sufficient information to direct high levels of gene expression in cells of retinal origin. Deletion or mutagenesis of an AP-1 motif present in this region caused 90-95% reduction in reporter gene expression. By gel mobility shift assay we demonstrated specific interactions between putative nuclear transcription factors and this AP-1 element. These findings indicate that the proximal AP-1 site in the human beta-PDE promoter is functionally relevant and necessary for transcriptional activation of this gene.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3',5'-Cyclic-GMP Phosphodiesterases / chemistry
  • 3',5'-Cyclic-GMP Phosphodiesterases / genetics*
  • Base Sequence
  • Binding Sites / genetics
  • Cell Line
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • DNA Primers / genetics
  • Gene Expression
  • Humans
  • Molecular Sequence Data
  • Mutagenesis
  • Nuclear Proteins / metabolism
  • Phosphoric Diester Hydrolases*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Protein Conformation
  • Retina / metabolism
  • Retinal Rod Photoreceptor Cells / metabolism*
  • Transcription Factor AP-1 / metabolism*
  • Transcriptional Activation*
  • Transfection

Substances

  • DNA Primers
  • Nuclear Proteins
  • Transcription Factor AP-1
  • Phosphoric Diester Hydrolases
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • PDE6B protein, human