Many behaviors, such as when we fall asleep or wake up, follow the rhythm of day and night. This is regulated in part by our ‘circadian clock’, which controls biological processes through the timed activation of hundreds of genes over the 24-hour day. In fruit flies, the proteins that form the core of the circadian clock activate and repress each other in such a way that their expression oscillates over a 24-hour cycle. During the late afternoon and early evening, the Clock protein initiates the production of proteins Period and Timeless: these two molecules then accumulate in the cell, and after binding to each other, they are transported into the nucleus. During the late night and early morning, this Period/Timeless complex inhibits the activity of Clock. After a delay, Period and Timeless are degraded. This allows Clock to be reactivated, restarting the cycle for the next day. Period is critical to help maintain the 24-hour oscillation shown by these proteins. A protein called Doubletime is responsible for making a number of chemical modifications on Period. It is unclear how these changes interact with each other, and how they influence the stability and function of Period when it is associated with Timeless. Here, Top et al. generate mutations in the fruit fly gene period to study these processes, and develop a new biomolecular technique to monitor the stability and activity of Period protein in insect cells grown in the laboratory. The experiments reveal new roles for the chemical changes made by Doubletime to Period. First, after Period associates with Timeless, Doubletime triggers certain modifications that lead to Period being able to inactivate Clock. Second, Doubletime makes another change in a nearby region of Period that results in the Period/Timeless complex being stabilized. Both sets of modifications help the complex to stay active and keep inhibiting Clock for long enough such that a 24-hour rhythm can be maintained. Finally, when Timeless is degraded, Period is released from the complex. At this time, the modifications made by Doubletime promote the degradation of Period, resetting the clock. Fruit flies with mutations that block this mechanism perceive the day as shorter. This shows that the smallest change to clock genes can disorganize behavior. Indeed in humans, health problems such as sleep or mental health disorders are associated with irregular circadian clocks. Understanding the biochemical mechanisms that keep the body clocks ticking could help to find new therapeutic targets for these conditions.
Doubletime kinase stabilizes the PER/TIM complex and regulates its transcriptional inhibition function to delay circadian transcriptional activity, helping sustain 24-hour periodicity in the circadian clock.
Phosphorylation of PERIOD Is Influenced by Cycling Physical Associations of DOUBLE-TIME, PERIOD, and TIMELESS in the Drosophila Clock
tim 0 ; tim DNLS -yfp flies show increased levels of PER and TIM
CK2 inhibits TIMELESS nuclear export and modulates CLOCK
CK2 inhibits TIMELESS nuclear export and modulates CLOCK transcriptional activity to regulate circadian rhythms
Real time, in vivo measurement of neuronal and peripheral clocks in Drosophila melanogaster
Real time, in vivo measurement of neuronal and peripheral clocks
Casein Kinase Iepsilon - an overview
Time Travels: A 40‐Year Journey from Drosophila's Clock Mutants to Human Circadian Disorders (Nobel Lecture) - Young - 2018 - Angewandte Chemie International Edition - Wiley Online Library
Figure 6 from A PER/TIM/DBT interval timer for Drosophila's
Frontiers PERIOD Phosphoclusters Control Temperature
Compared with the other alleles, DBT ar and DBT K38R extend the
PERIOD phosphorylation leads to feedback inhibition of CK1 activity to control circadian period
NAD+ Controls Circadian Reprogramming through PER2 Nuclear
Integration of Circadian Clock Information in the Drosophila
