A novel interaction between Glycogen Synthase Kinase-3α (GSK-3α) and the scaffold protein Receptor for Activated C-Kinase 1 (RACK1) regulates the circadian clock

Glycogen synthase kinase-3α (GSK-3α) and GSK-3β are intracellular kinases with largely redundant functions. However, the deletion of each GSK-3 isoform in the mouse has distinct consequences, suggesting that these related enzymes also have non-overlapping isoform-specific functions. A yeast two-hybrid screen for GSK-3α interacting partners revealed an interaction with the Receptor for Activated C-Kinase 1 (RACK1). We confirm this interaction in mammalian cells, and provide evidence that RACK1 does not interact with GSK-3β. Structure-function analyses revealed that WD repeats 5-6 are required to interact with GSK-3α. Furthermore, this interaction is independent of GSK-3α activity. Finally, our data show that the GSK-3α-RACK1 interaction is necessary for regulating the circadian clock in mammalian cells. In summary, our data provides a mechanistic link between GSK-3 and RACK-1 in the regulation of the circadian clock, and demonstrates that this effect is specific to the GSK-3α isoform.