Rhythmic regulation of Ca2+ signalling networks

The circadian clock is the internal timekeeper of plants. This clock regulates most aspects of plant physiology providing considerable competitive advantage. We are investigating the role for oscillations in the cytosolic free Ca2+ concentration ([Ca2+]cyt) in the circadian control of cellular physiology. We have previously demonstrated that circadian oscillations of [Ca2+]cyt encode photoperiodic information but the precise role of circadian [Ca2+]cyt oscillations remain obscure. We have been taking a systems wide approach to determine the origin and function of circadian oscillations of [Ca2+]cyt. Using pharmacology, bioinformatics and biochemical tools we have new evidence that oscillations of [Ca2+]cyt are generated by the small signalling intermediate, cADPR. Positioning the oscillations of [Ca2+]cyt with respect to the molecular oscillator using reverse genetics indicates that [Ca2+]cyt is an output of the clock. Using a whole genome transcriptional profile we have identified over 1800 circadian-regulated transcripts, many of which encode for Ca2+ signalling elements. The function of circadian-regulated transcripts encoding signalling components is being investigated by reverse genetic screens with automated imaging. Using our extensive data sets describing the circadian regulation of [Ca2+]cyt in different backgrounds and conditions we have constructed a mathematical model. This is being validated using mutant analysis and refined by introducing complexity to the model. Our data and models suggest that [Ca2+]cyt acts an output of the clock that regulates diverse aspects of physiology and has the potential to form a feedback loop with the molecular components of the oscillator.