DOI: 10.1042/bj3370575. Role of phosphatidylinositol 3,4,5-trisphosphate in regulating the activity and localization of 3-phosphoinositide-dependent protein kinase-1

Role of phosphatidylinositol 3,4,5-trisphosphate in regulating the activity and localization of 3-phosphoinositide-dependent protein kinase-1

10.1042/bj3370575

Crossref journal-article

Portland Press Ltd.

Biochemical Journal (288)

Abstract

3-Phosphoinositide-dependent protein kinase-1 (PDK1) interacts stereoselectively with the d-enantiomer of PtdIns(3,4,5)P3 (KD 1.6 nM) and PtdIns(3,4)P2 (KD 5.2 nM), but binds with lower affinity to PtdIns3P or PtdIns(4,5)P2. The binding of PtdIns(3,4,5)P3 to PDK1 was greatly decreased by making specific mutations in the pleckstrin homology (PH) domain of PDK1 or by deleting it. The same mutations also greatly decreased the rate at which PDK1 activated protein kinase Bα (PKBα) in vitro in the presence of lipid vesicles containing PtdIns(3,4,5)P3, but did not affect the rate at which PDK1 activated a PKBα mutant lacking the PH domain in the absence of PtdIns(3,4,5)P3. When overexpressed in 293 or PAE cells, PDK1 was located at the plasma membrane and in the cytosol, but was excluded from the nucleus. Mutations that disrupted the interaction of PtdIns(3,4,5)P3 or PtdIns(4,5)P2 with PDK1 abolished the association of PDK1 with the plasma membrane. Growth-factor stimulation promoted the translocation of transfected PKBα to the plasma membrane, but had no effect on the subcellular distribution of PDK1 as judged by immunoelectron microscopy of fixed cells. This conclusion was also supported by confocal microscopy of green fluorescent protein–PDK1 in live cells. These results, together with previous observations, indicate that PtdIns(3,4,5)P3 plays several roles in the PDK1-induced activation of PKBα. First, it binds to the PH domain of PKB, altering its conformation so that it can be activated by PDK1. Secondly, interaction with PtdIns(3,4,5)P3 recruits PKB to the plasma membrane with which PDK1 is localized constitutively by virtue of its much stronger interaction with PtdIns(3,4,5)P3 or PtdIns(4,5)P2. Thirdly, the interaction of PDK1 with PtdIns(3,4,5)P3 facilitates the rate at which it can activate PKB.

Bibliography

CURRIE, R. A., WALKER, K. S., GRAY, A., DEAK, M., CASAMAYOR, A., DOWNES, C. P., COHEN, P., ALESSI, D. R., & LUCOCQ, J. (1999). Role of phosphatidylinositol 3,4,5-trisphosphate in regulating the activity and localization of 3-phosphoinositide-dependent protein kinase-1. Biochemical Journal, 337(3), 575â583.

Authors 9

Richard A. CURRIE (first)
Kay S. WALKER (additional)
Alex GRAY (additional)
Maria DEAK (additional)
Antonio CASAMAYOR (additional)
C. Peter DOWNES (additional)
Philip COHEN (additional)
Dario R. ALESSI (additional)
John LUCOCQ (additional)

References 0 Referenced 288

None

Dates

Type	When
Created	10 years ago (Aug. 10, 2015, 6:04 p.m.)
Deposited	3 years, 9 months ago (Nov. 23, 2021, 7:42 p.m.)
Indexed	36 minutes ago (Sept. 6, 2025, 9:09 a.m.)
Issued	26 years, 7 months ago (Jan. 25, 1999)
Published	26 years, 7 months ago (Jan. 25, 1999)
Published Online	26 years, 7 months ago (Jan. 25, 1999)
Published Print	26 years, 7 months ago (Feb. 1, 1999)

Funders 0

None

BibTeX

@article{CURRIE_1999, title={Role of phosphatidylinositol 3,4,5-trisphosphate in regulating the activity and localization of 3-phosphoinositide-dependent protein kinase-1}, volume={337}, ISSN={1470-8728}, url={http://dx.doi.org/10.1042/bj3370575}, DOI={10.1042/bj3370575}, number={3}, journal={Biochemical Journal}, publisher={Portland Press Ltd.}, author={CURRIE, Richard A. and WALKER, Kay S. and GRAY, Alex and DEAK, Maria and CASAMAYOR, Antonio and DOWNES, C. Peter and COHEN, Philip and ALESSI, Dario R. and LUCOCQ, John}, year={1999}, month=jan, pages={575–583} }

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