Dynamic sequestration of the recycling compartment by classical protein kinase C

Jolanta Idkowiak-Baldys, Kevin P. Becker, Kazuyuki Kitatani, Yusuf A. Hannun

Research output: Contribution to journalArticlepeer-review

43 Citations (Scopus)

Abstract

It has been previously shown that upon sustained stimulation (30-60 min) with phorbol esters, protein kinase C (PKC) α and βII 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.

Original languageEnglish
Pages (from-to)22321-22331
Number of pages11
JournalJournal of Biological Chemistry
Volume281
Issue number31
DOIs
Publication statusPublished - 2006 Aug 4
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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