Activation of the essential kinase PDK1 by phosphoinositide-driven trans-autophosphorylation

3-phosphoinositide-dependent kinase 1 (PDK1) is an essential serine/threonine protein kinase, which plays a crucial role in cell growth and proliferation. It is often referred to as a ‘master’ kinase due to its ability to activate at least 23 downstream protein kinases implicated in various signaling pathways. In this study, we have elucidated the mechanism of phosphoinositide-driven PDK1 auto-activation. We show that PDK1 trans -autophosphorylation is mediated by a PIP 3 -mediated face-to-face dimer. We report regulatory motifs in the kinase-PH interdomain linker that allosterically activate PDK1 autophosphorylation via a linker-swapped dimer mechanism. Finally, we show that PDK1 is autoinhibited by its PH domain and that positive cooperativity of PIP 3 binding drives switch-like activation of PDK1. These results imply that the PDK1-mediated activation of effector kinases, including Akt, PKC, Sgk, S6K and RSK, many of whom are not directly regulated by phosphoinositides, is also likely to be dependent on PIP 3 or PI(3,4)P 2 . The essential protein kinase PDK1 is activated by phospoinositide-mediated dimerization and trans -autophosphorylation. Here, the authors show that in the absence of PIP 3 or PI(3,4)P 2 phosphoinositides, PDK1 is maintained in an inactive, autoinhibited conformation in the cytosol.