Abstract
In the seasonal cycle of the silkworm Bombyx mori, an ATPase called esterase A4 (EA4) is thought to measure time interval as a diapause-duration timer. To address the mechanism by which EA4 measures the time, we present a simple EA4 screening method. By the method, EA4 activity can be assessed with a short incubation at 25°C without the need to purify the enzyme further. The method largely overcomes methodological problems that remain unanswered. Besides, the results obtained by the method establish that the time measurement is based on the moment when the time-holding factor(s) PIN (peptidyl-inhibitory needle) is removed. EA4 may originally be a complex with PIN, and external time cues such as winter cold may induce the dissociation of the complex, which in turn results in the timer activation of EA4.
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