Zn²⁺ influx activates ERK and Akt signaling pathways

Zinc (Zn²⁺) is an essential metal in biology, and its bioavailability is highly regulated. Many cell types exhibit fluctuations in Zn²⁺ that appear to play an important role in cellular function. However, the detailed molecular mechanisms by which Zn²⁺ dynamics influence cell physiology remain enigmatic. Here, we use a combination of fluorescent biosensors and cell perturbations to define how changes in intracellular Zn²⁺ impact kinase signaling pathways. By simultaneously monitoring Zn²⁺ dynamics and kinase activity in individual cells, we quantify changes in labile Zn²⁺ and directly correlate changes in Zn²⁺ with ERK and Akt activity. Under our experimental conditions, Zn²⁺ fluctuations are not toxic and do not activate stress-dependent kinase signaling. We demonstrate that while Zn²⁺ can nonspecifically inhibit phosphatases leading to sustained kinase activation, ERK and Akt are predominantly activated via upstream signaling and through a common node via Ras. We provide a framework for quantification of Zn²⁺ fluctuations and correlate these fluctuations with signaling events in single cells to shed light on the role that Zn²⁺ dynamics play in healthy cell signaling.