Protein kinase C(alpha) actively downregulates through caveolae-dependent traffic to an endosomal compartment.
Receptor desensitization occurs through receptor internalization and targeting to endosomes, a prerequisite for sorting and degradation. Such trafficking processes may not be restricted to membrane associated receptors but may also play an important role in the downregulation of cytoplasmic transducers such as protein kinase C (PKC). It is demonstrated here that acute TPA exposure induces the transport of activated PKC(alpha) from the plasma membrane to endosomes. This process requires PKC activity and catalytically competent PKC can even promote a similar process for a truncated regulatory domain PKC(&agr;) protein. It is established that PKC(&agr;) is targeted to the endosome compartment as an active kinase, where it colocalizes with annexin I, a substrate of PKC. Thus, PKC(alpha) downregulation shares features with plasma membrane associated receptor sorting and degradation. However, it is shown that PKC(&agr;) delivery to the endosome compartment is not a Rab5 mediated process in contrast to the well characterised internalisation of the transferrin receptor. An alternative route for PKC(alpha) is evidenced by the finding that the cholesterol binding drugs nystatin and filipin, known to inhibit caveolae mediated trafficking, are able to block PKC(alpha) traffic and down regulation. Consistent with this, the endosomes where PKC(alpha) is found also contain caveolin. It is concluded that the initial step in desensitisation of PKC(alpha) involves active delivery to endosomes via a caveolae mediated process.
Prevostel
C
C
Protein Phosphorylation Laboratory, Imperial Cancer Research Fund, London WC2a 3PX, UK.
Alice
V
V
Joubert
D
D
Parker
P J
PJ
D000818:Animals; D002451:Cell Compartmentation; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D011992:Endosomes / Q000187:drug effects / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry / Q000187:drug effects / Q000502:physiology; D008856:Microscopy, Fluorescence; D009761:Nystatin / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011493:Protein Kinase C / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D051571:Protein Kinase C-alpha; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D011990:Receptors, Transferrin / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection / Q000235:genetics; D020696:rab5 GTP-Binding Proteins / Q000187:drug effects / Q000378:metabolism
D007527:Isoenzymes; D008565:Membrane Proteins; D011990:Receptors, Transferrin; D009761:Nystatin; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D020696:rab5 GTP-Binding Proteins; D013755:Tetradecanoylphorbol Acetate