Cdk5 regulates IP3R1-mediated Ca²⁺ dynamics and Ca²⁺-mediated cell proliferation

Loss of cyclin-dependent kinase 5 (Cdk5) in the mitochondria-associated endoplasmic reticulum (ER) membranes (MAMs) increases ER-mitochondria tethering and ER Ca²⁺ transfer to the mitochondria, subsequently increasing mitochondrial Ca²⁺ concentration ([Ca²⁺]_mt). This suggests a role for Cdk5 in regulating intracellular Ca²⁺ dynamics, but how Cdk5 is involved in this process remains to be explored. Using ex vivo primary mouse embryonic fibroblasts (MEFs) isolated from Cdk5^-/- mouse embryos, we show here that loss of Cdk5 causes an increase in cytosolic Ca²⁺concentration ([Ca²⁺]_cyt), which is not due to reduced internal Ca²⁺ store capacity or increased Ca²⁺ influx from the extracellular milieu. Instead, by stimulation with ATP that mediates release of Ca²⁺ from internal stores, we determined that the rise in [Ca²⁺]_cyt in Cdk5^-/- MEFs is due to increased inositol 1,4,5-trisphosphate receptor (IP3R)-mediated Ca²⁺ release from internal stores. Cdk5 interacts with the IP3R1 Ca²⁺ channel and phosphorylates it at Ser₄₂₁. Such phosphorylation controls IP3R1-mediated Ca²⁺ release as loss of Cdk5, and thus, loss of IP3R1 Ser₄₂₁ phosphorylation triggers an increase in IP3R1-mediated Ca²⁺ release in Cdk5^-/- MEFs, resulting in elevated [Ca²⁺]_cyt. Elevated [Ca²⁺]_cyt in these cells further induces the production of reactive oxygen species (ROS), which upregulates the levels of Nrf2 and its targets, Prx1 and Prx2. Cdk5^-/- MEFs, which have elevated [Ca²⁺]_cyt, proliferate at a faster rate compared to wt, and Cdk5^-/- embryos have increased body weight and size compared to their wt littermates. Taken together, we show that altered IP3R1-mediated Ca²⁺ dynamics due to Cdk5 loss correspond to accelerated cell proliferation that correlates with increased body weight and size in Cdk5^-/- embryos.