Abstract
Light is the dominant environmental cue that provides temporal information to circadian pacemakers. In Drosophila melanogaster some period gene mutants have altered free-running circadian periods but entrain to 24 h light-dark cycles. To address the mechanism of light entrainment in Drosophila, we examined the effects of constant light on the period gene (per) and timeless gene (tim) products in wild-type and perS flies. The results indicate that light affects three features of the PER-TIM program: PER and TIM phosphorylation, PER and TIM accumulation, and per and tim RNA cycling. A post-transcriptional effect on the PER-TIM complex is the likely primary clock target, which then delays the subsequent decrease in per and tim RNA levels. This is consistent with a negative feedback loop, in which the PER-TIM complex contributes to the decrease in per and tim RNA levels, presumably at the transcriptional level. There are enhanced constant light effects on the perS mutant, which further support negative feedback as well as support its importance to entrainment of these flies to a 24 h cycle, far from their intrinsic period of 19 h. The perS mutant leads to a truncated protein accumulation phase and a subsequent premature perS RNA increase. A standard 24 h light-dark cycle delays the negative feedback circuit and extends the RNA and protein profiles, compensating for the accelerated RNA increase and restoring the rhythms to wild-type-like periodicity.
