1. Chemotactic activation of Dictyostelium AGC-family kinases AKT and PKBR1 requires separate but coordinated functions of PDK1 and TORC2.
- Author
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Liao XH, Buggey J, and Kimmel AR
- Subjects
- 3-Phosphoinositide-Dependent Protein Kinases, Amino Acid Motifs, Animals, Chemotactic Factors pharmacology, Cyclic AMP pharmacology, Dictyostelium drug effects, Enzyme Activation drug effects, Folic Acid pharmacology, Lipid Metabolism drug effects, Phosphatidylinositol 3-Kinases metabolism, Phosphatidylinositol Phosphates metabolism, Phosphorylation drug effects, Signal Transduction drug effects, Substrate Specificity drug effects, Chemotaxis drug effects, Dictyostelium cytology, Dictyostelium enzymology, Protein Serine-Threonine Kinases metabolism, Proto-Oncogene Proteins c-akt metabolism, Protozoan Proteins metabolism, Trans-Activators metabolism
- Abstract
Protein kinases AKT and PKBR1 of Dictyostelium belong to the AGC protein kinase superfamily. AKT and PKBR1 are phosphorylated at similar sites by phosphoinositide-dependent kinase 1 (PDK1) and TORC2 kinases; however, they have different subcellular localizing domains. AKT has a phosphoinositide 3-kinase (PI3K)/phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P(3)]-regulated PH (pleckstrin homology) domain whereas PKBR1 is myristoylated and persistently membrane localized. Using strains defective for PI3K/PtdIns(3,4,5)P(3)-, PDK1- and TORC2-signaling or strains that express phospho-site mutants of AKT and PKBR1, we dissect the different roles of PI3K/PtdIns(3,4,5)P(3), PDK1 and TORC2. We show that activation of AKT and PKBR1 requires PDK1-site phosphorylation, but that phosphorylation by TORC2 is insufficient for AKT or PKBR1 activation. However, PDK1-site phosphorylation is dependent on phosphorylation by TORC2, which suggests that there is regulatory coordination among PDK1, TORC2 and their phospho-site targets. This defines a separate input for signaling in control of chemotaxis and dependency on PDK1 function. We also demonstrate that PDK1 in Dictyostelium functions independently of PI3K/PtdIns(3,4,5)P(3). Finally, we show that AKT and PKBR1 exhibit substrate selectivity and identify two novel lipid-interacting proteins preferentially phosphorylated by AKT. Despite certain similarities, AKT and PKBR1 have distinct regulatory paths that impact activation and effector targeting, with PDK1 serving a central role.
- Published
- 2010
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