An Inhibitor of Protein Phosphorylation Stops the Circadian Oscillator and Blocks Light-Induced Phase Shifting in Gonyaulax polyedra

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Abstract
The expression of circadian rhythmicity in Gonyaulax polyedra is strikingly altered by an inhibitor of protein phosphorylation. The effects of 6-dimethylaminopurine (6-DMAP), known to reversibly block cell division in many systems through inhibition of protein kinase activity, are described here for Gonyaulax. Its action appears to be exclusively tonic in nature; in cells continuously exposed, the period is lengthened in a concentration-dependent fashion. Shorter treatments at a higher concentra tion of 6-DMAP (5 mM) apparently stop the circadian oscillator, but reversibly so, since the rhythm resumes after drug removal with a phase delay approximately equal to the duration of the treatment. Pulses of the inhibitor are effective in causing phase delays at all times of the circadian cycle. In addition, 6-DMAP completely blocks light-induced phase advances and is effective in inhibiting many Gonyaulax protein kinases in vitro.