Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C

It has been previously shown that upon sustained stimulation (30-60 min) with phorbol esters, protein kinase C (PKC)α andβII become sequestered in a juxtanuclear region, the pericentrion. The activation of PKC also results in sequestration of transferrin, suggesting a role for PKC in regulating endo...

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Bibliographic Details
Published in:The Journal of biological chemistry 2006-08, Vol.281 (31), p.22321-22331
Main Authors: Idkowiak-Baldys, Jolanta, Becker, Kevin P., Kitatani, Kazuyuki, Hannun, Yusuf A.
Format: Article
Language:English
Subjects:
CD59 Antigens - metabolism
Cell Compartmentation - drug effects
Cell Line
Clathrin - metabolism
Endocytosis
Humans
Intracellular Membranes - drug effects
Protein Kinase C - metabolism
Protein Kinase C - physiology
Protein Kinase C-alpha - metabolism
Protein Kinase C-alpha - physiology
Temperature
Tetradecanoylphorbol Acetate - pharmacology
Transfection
Transferrin - metabolism
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Summary:It has been previously shown that upon sustained stimulation (30-60 min) with phorbol esters, protein kinase C (PKC)α andβII become sequestered in a juxtanuclear region, the pericentrion. The activation of PKC also results in sequestration of transferrin, suggesting a role for PKC in regulating endocytosis and sequestration of recycling components. In this work we characterize the pericentrion as a PKC-dependent subset of the recycling compartment. We demonstrate that upon sustained stimulation of PKC, both protein (CD59, caveolin) and possibly also lipid (Bodipy-GM1) cargo become sequestered in a PKC-dependent manner. This sequestration displayed a strict temperature requirement and was inhibited below 32 °C. Treatment of cells with phorbol myristate acetate for 60 min led to the formation of a distinct membrane structure. PKC sequestration and pericentrion formation were blocked by hypertonic sucrose as well as by potassium depletion (inhibitors of clathrin-dependent endocytosis) but not by nystatin or filipin, which inhibit clathrin-independent pathways. Interestingly, it was also observed that some molecules that internalize through clathrin-independent pathways (CD59, Bodipy-GM1, caveolin) also sequestered to the pericentrion upon sustained PKC activation, suggesting that PKC acted distal to the site of internalization of endocytic cargo. Together these results suggest that PKC regulates sequestration of recycling molecules into this compartment, the pericentrion.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M512540200