Isolation and initial characterization of the 5' flanking region of the human and murine cyclic guanosine monophosphate-phosphodiesterase beta-subunit genes

Invest Ophthalmol Vis Sci. 1996 Mar;37(4):551-60.


Purpose: As an initial approach to study the mechanisms that direct photoreceptor-specific expression of the rod cyclic guanosine monophosphate-phosphodiesterase beta-subunit (beta-PDE) gene, the 5' flanking regions of the human and mouse genes were cloned and analyzed.

Methods: Genomic libraries were screened and clones containing the 5' upstream region of the beta-PDE gene were isolated and sequenced. Primer extension and ribonuclease protection assays were used to determine the transcription initiation sites. Sequences were compared using dot-matrix analysis and nucleotide alignment to determine potential regulatory elements that have been conserved through evolution. DNA-protein interactions were examined using DNAse I footprinting.

Results: The beta-PDE gene 5' sequence contains two distinct transcription start sites and lacks a TATA box. A stretch of approximately 30 nucleotides just upstream of the first transcribed nucleotide is strongly conserved in both species. This sequence contains a TATA-like element and a -CTAATC- motif previously described in other photoreceptor-specific genes. A highly-conserved AP-1 element, the recognition site for members of the jun and the fos oncoproteins family, is also present in this proximal region. DNAse I footprinting revealed an array of retinal proteins binding to these elements.

Conclusions: The beta-PDE 5' region features match those of a highly tissue-specific gene in which factors restricted to the retina might play a role in gene activation. Elements conserved through evolution in the human and mouse sequences were found and analyzed as potential cis-acting elements. The availability of the human beta-PDE 5' upstream sequence will allow patients with retinal degeneration to be screened for possible mutations in these control sequences.

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 / genetics*
  • 3',5'-Cyclic-GMP Phosphodiesterases / isolation & purification
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Conserved Sequence
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • DNA Primers / chemistry
  • Female
  • Gene Library
  • Genes, Regulator / genetics
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Phosphoric Diester Hydrolases*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • Retina / enzymology*
  • Sequence Analysis, DNA
  • Sequence Homology, Nucleic Acid
  • TATA Box / genetics
  • Transcription, Genetic / genetics


  • DNA Primers
  • Phosphoric Diester Hydrolases
  • 3',5'-Cyclic-GMP Phosphodiesterases
  • Cyclic Nucleotide Phosphodiesterases, Type 6
  • PDE6B protein, human
  • Pde6b protein, mouse

Associated data

  • GENBANK/U31760
  • GENBANK/U31761