PMID: 8595955Mar 1, 1996Paper

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

Investigative Ophthalmology & Visual Science
A Di PoloD B Farber

Abstract

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. 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. 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 foot...Continue Reading

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