Characterization of canine photoreceptor phosducin cDNA and identification of a sequence variant in dogs with photoreceptor dysplasia

Gene. 1998 Jul 30;215(2):231-9. doi: 10.1016/s0378-1119(98)00310-2.

Abstract

Photoreceptor dysplasia (pd) is an autosomal recessive disease of miniature schnauzer dogs causing retinal degeneration. The disease is a homologue of retinitis pigmentosa, a group of genetically heterogeneous diseases, causing blindness in humans. A subtraction library was prepared from retinas of pd affected and age-matched normal control dogs to isolate de novo candidate genes for further examination. From the subtraction library, cDNA for phosducin (PDC), a member of the phototransduction pathway, was isolated as a transcript expressed at a higher level in the affected retina. First, the normal canine PDC cDNA was characterized to evaluate the PDC gene in the pd-affected retina. The characterized region of normal PDC cDNA spans 1258 nucleotides (nt) that include 738 nt of coding sequence predicted to encode a protein (Mr=28 209) of 245 amino acids (aa). Over the coding region, PDC shares 86-95% nt sequence identity and 90-95% identity in the deduced aa sequence with homologous mammalian sequences. A major transcript (1.9 kb) was observed only in retina by Northern analysis, but low levels of transcript were detected in brain, liver and kidney by reverse transcription and polymerase chain reaction. Retinal immunocytochemistry showed that PDC was detected only in rod photoreceptors, mainly in the inner segment and perinuclear region. By Northern blot analysis, increased PDC expression was observed in pre-degenerate affected retina relative to the age-matched normal. In pd- affected miniature schnauzer pedigree, a missense mutation was detected in codon 82 (CGA to GGA) that would create a non-conservative substitution (Arg to Gly) in close vicinity to the residue (Glu 85) which directly interacts with the betagamma-subunits of transducin. Only pd-affected dogs were found to be homozygous for the mutant allele, and none among 48 dogs tested from 20 other dog breeds had this allele, suggesting that the mutation could be causally associated with pd in miniature schnauzers. However, since some affected dogs are heterozygous for the mutant allele, and some are homozygous for the wild-type allele, this putative PDC missense mutation, if it is indeed a disease causing mutation, does not account entirely for the genetics of inherited retinal degeneration in the miniature schnauzer breed.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • DNA Primers
  • DNA, Complementary
  • Dog Diseases / genetics*
  • Dog Diseases / pathology
  • Dogs / genetics*
  • Eye Proteins / biosynthesis
  • Eye Proteins / chemistry
  • Eye Proteins / genetics*
  • GTP-Binding Protein Regulators
  • Genes, Recessive
  • Genetic Carrier Screening
  • Genetic Variation*
  • Molecular Sequence Data
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics*
  • Photoreceptor Cells / pathology*
  • Polymerase Chain Reaction
  • Restriction Mapping
  • Retina / pathology
  • Retinal Degeneration / genetics
  • Retinal Degeneration / pathology
  • Retinal Degeneration / veterinary*
  • Species Specificity

Substances

  • DNA Primers
  • DNA, Complementary
  • Eye Proteins
  • GTP-Binding Protein Regulators
  • Phosphoproteins
  • phosducin

Associated data

  • GENBANK/AF046874