Influence of various domains of protein kinase C epsilon on its PMA-induced translocation from the Golgi to the plasma membrane.
Subcellular redistribution (translocation) was initiated by treatment of NIH 3T3 cells overexpressing different epitope-tagged fragments of PKC epsilon with PMA, and was analyzed by immunocytochemistry. The PMA-induced translocation of holo PKC epsilon, as well as fragments epsilon 2 (zinc finger domain + pseudosubstrate domain) and epsilon 7 (zinc finger domain + hinge region) from the Golgi to the plasma membrane was rapid (<10 min), while translocation of fragment epsilon 3 (zinc finger domain) was much slower (30-60 min). These results, combined with results of studies carried out at 20 degrees C to inhibit exocytotic vesicle traffic, indicated that PMA-induced translocation from the Golgi to the plasma membrane may proceed by two distinct mechanisms: a rapid, vesicle independent process noted with holo PKC epsilon (which requires the presence of the pseudosubstrate and/or hinge regions), and a slow, vesicle-dependent pathway observed with the zinc finger fragment.
Lehel
C
C
Laboratory of Cellular Oncology, NCI, NIH, Bethesda, Maryland 20892, USA.
Oláh
Z
Z
Petrovics
G
G
Jakab
G
G
Anderson
W B
WB
D016475:3T3 Cells; D000085:Acetates / Q000494:pharmacology; D019342:Acetic Acid; D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D005089:Exocytosis; D015870:Gene Expression / Q000187:drug effects; D006056:Golgi Apparatus / Q000201:enzymology*; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D016296:Mutagenesis; D011493:Protein Kinase C / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D016324:Sequence Tagged Sites; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection; D016335:Zinc Fingers
D000085:Acetates; D007527:Isoenzymes; D011994:Recombinant Proteins; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D019342:Acetic Acid