Photoneural control of the synthesis and phosphorylation of pineal MEKA (phosducin).
MEKA is an acidic 33-kilodalton phosphoprotein found in the retina and pineal gland. It is of interest because it forms a cytoplasmic heterotrimer with the beta gamma-complex of GTP-binding regulatory proteins (G proteins). Accordingly, MEKA may play a role in signal transduction. MEKA is phosphorylated on Ser73 by cAMP-dependent protein kinase. In the present report, MEKA was studied using an antiserum (Anti-32) against MEKA65-96, which can be used to estimate total MEKA and the phosphorylation state of MEKA. It was confirmed that MEKA is rapidly phosphorylated by adrenergic stimulation of pineal glands in organ culture. In addition, total (dephosphorylated) MEKA was observed to increase after a 6-h treatment with norepinephrine or (Bu)2 cAMP, an effect which was dependent upon new protein synthesis. In in vivo studies, it was found that the total amount of MEKA and MEKA phosphorylation were increased at night in the dark, a time when the pineal gland is adrenergically stimulated. The high level of phosphorylation was rapidly reduced when animals were exposed to light, which blocks neural stimulation of the gland. This report provides the first in vivo evidence that MEKA phosphorylation is under physiological control, and that MEKA synthesis is controlled by an adrenergic----cAMP mechanism which requires protein synthesis.
Schaad
N C
NC
Section on Neuroendocrinology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Shinohara
T
T
Abe
T
T
Klein
D C
DC
D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002940:Circadian Rhythm; D003513:Cycloheximide / Q000494:pharmacology; D005136:Eye Proteins / Q000096:biosynthesis / Q000378:metabolism*; D020555:GTP-Binding Protein Regulators; D008027:Light; D008297:Male; D009638:Norepinephrine / Q000494:pharmacology*; D009924:Organ Culture Techniques; D010750:Phosphoproteins / Q000096:biosynthesis / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism; D010870:Pineal Gland / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
D005136:Eye Proteins; D020555:GTP-Binding Protein Regulators; D010750:Phosphoproteins; C065631:phosducin; D010768:Phosphoserine; D003994:Bucladesine; D003513:Cycloheximide; D009638:Norepinephrine