Functional contributions of ion fluxes to cellular circadian organisation

Our recent work revealed that circadian rhythms exist in the intracellular concentrations of key ions fundamental to cellular life: magnesium and potassium. We now propose a creative research programme around the central hypothesis that fluxes of ions provide key regulatory functions in the circadian organisation of the cellular landscape. Objective 1 is to delineate ion concentration rhythms at the sub-cellular level and use multi-omics techniques to reveal how these integrate into the rhythmic transcriptome, proteome, and metabolome over the diurnal and circadian cycle. Objective 2 will reveal the full cellular magnesium transport machinery of a eukaryotic cell and use gene editing and live cell imaging to visualise the dynamics subcellular magnesium fluxes, as well as long-term adaptation experiments to identify those cellular functions that are sensitive to magnesium rhythms. Objective 3 will investigate the functional effects of potassium concentration rhythms in progression of the cell cycle, in gatekeeping glycolysis and primary metabolism, and in facilitating regulated cellular proteostasis throughout the cell and circadian cycles. Overall, the program will deliver systems level understanding of spatiotemporal ion fluxes and their key roles in the rhythmic orchestration of cellular properties that are fundamental to the health of a eukaryotic cell.