Rat pineal gland phosducin

cDNA isolation, nucleotide sequence, and chromosomal assignment in the mouse

Cheryl M. Craft, Richard N. Lolley, Michael F Seldin, Rehwa H. Lee

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

The pineal gland contains a soluble phosphoprotein, phosducin, which is homologous to that of retinal photoreceptors. Phosducin has been shown to bind the β, γ subunits of the retinal G-protein transducin. Retinal phosducin has been cloned and now we report a rat pineal cDNA encoding phosducin. A 1217-nucleotide cDNA was isolated from a rat pineal library by DNA-DNA hybridization with a polymerase chain reaction-amplified cDNA of bovine retina mRNA for phosducin. Northern blot analysis demonstrates that the mRNA for phosducin is approximately 1.3 kb in both rat pineal and rat retina. The translated mRNA from rat pineal encodes a protein with 246 amino acids, compared to the 245 amino acids of bovine retina phosducin. The predicted molecular weight of rat pineal phosducin is 28,201. Immunoblot analysis with affinity-purified antibodies against bovine retina phosducin identify a single immunoreactive protein of approximately 33 kDa in both rat retina and rat pineal. The amino acid sequence of rat pineal phosducin is homologous to that of bovine retina phosducin, revealing 89% identity and another 5.7% similarity. Both rat pineal and bovine retina phosducins are acidic proteins with pIs of 4.3 and 4.5, respectively. The translated protein lacks hydrophobic domains that would suggest an integral membrane protein. Rat pineal phosducin has a single consensus phosphorylation domain for protein kinase A that is nearly identical to that of retinal phosducin, which is phosphorylated by protein kinase A in situ. Rat phosducin also contains three potential phosphorylation domains for protein kinase C and nine for casein kinase II as well as a predicted site for N-glycosylation. The cDNA encoding phosducin was used to localize the gene within a linkage group to a large segment of mouse chromosome 1 in a conserved region with the long arm of human chromosome 1 with a panel of DNA samples from an interspecific cross. In keeping with a proposed role of retinal phosducin in down-regulation of the photo-transduction cascade, a modulatory role in signal transduction is proposed for pineal phosducin.

Original languageEnglish (US)
Pages (from-to)400-409
Number of pages10
JournalGenomics
Volume10
Issue number2
DOIs
StatePublished - 1991

Fingerprint

Pineal Gland
Complementary DNA
Retina
phosducin
Chromosomes, Human, Pair 1
Cyclic AMP-Dependent Protein Kinases
Messenger RNA
Proteins
Phosphorylation
Transducin
Casein Kinase II
Amino Acids
Vertebrate Photoreceptor Cells
Antibody Affinity
Phosphoproteins
DNA
Human Chromosomes

ASJC Scopus subject areas

  • Genetics

Cite this

Rat pineal gland phosducin : cDNA isolation, nucleotide sequence, and chromosomal assignment in the mouse. / Craft, Cheryl M.; Lolley, Richard N.; Seldin, Michael F; Lee, Rehwa H.

In: Genomics, Vol. 10, No. 2, 1991, p. 400-409.

Research output: Contribution to journalArticle

Craft, Cheryl M. ; Lolley, Richard N. ; Seldin, Michael F ; Lee, Rehwa H. / Rat pineal gland phosducin : cDNA isolation, nucleotide sequence, and chromosomal assignment in the mouse. In: Genomics. 1991 ; Vol. 10, No. 2. pp. 400-409.
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