Clock and riboswitch control of THIC in tandem are essential for appropriate gauging of TDP levels under light/dark cycles in Arabidopsis.

Metabolic homeostasis is regulated by enzyme activities, but the importance of regulating their corresponding coenzyme levels is unexplored. The organic coenzyme thiamine diphosphate (TDP) is suggested to be supplied as needed and controlled by a riboswitch-sensing mechanism in plants through the circadian-regulated THIC gene. Riboswitch disruption negatively impacts plant fitness. A comparison of riboswitch-disrupted lines to those engineered for enhanced TDP levels suggests that time-of-day regulation of THIC expression particularly under light/dark cycles is crucial. Altering the phase of THIC expression to be synchronous with TDP transporters disrupts the precision of the riboswitch implying that temporal separation of these processes by the circadian clock is important for gauging its response. All defects are bypassed by growing plants under continuous light conditions, highlighting the need to control levels of this coenzyme under light/dark cycles. Thus, consideration of coenzyme homeostasis within the well-studied domain of metabolic homeostasis is highlighted.

The coenzyme thiamine diphosphate displays a daily rhythm in the Arabidopsis nucleus.

In plants, metabolic homeostasis-the driving force of growth and development-is achieved through the dynamic behavior of a network of enzymes, many of which depend on coenzymes for activity. The circadian clock is established to influence coordination of supply and demand of metabolites. Metabolic oscillations independent of the circadian clock, particularly at the subcellular level is unexplored. Here, we reveal a metabolic rhythm of the essential coenzyme thiamine diphosphate (TDP) in the Arabidopsis nucleus. We show there is temporal separation of the clock control of cellular biosynthesis and transport of TDP at the transcriptional level. Taking advantage of the sole reported riboswitch metabolite sensor in plants, we show that TDP oscillates in the nucleus. This oscillation is a function of a light-dark cycle and is independent of circadian clock control. The findings are important to understand plant fitness in terms of metabolite rhythms.