Polarized exocytosis in MDCK cells is regulated by phosphorylation.
Protein phosphorylation and dephosphorylation systems modulate many cellular activities and have recently been implicated in the in vitro transport of newly synthesized proteins. Here we show that polarized transport from the Golgi to the plasma membrane in intact MDCK cells is regulated by phosphorylation-dephosphorylation. Transport is inhibited by the phosphatase inhibitor okadaic acid and is stimulated by the kinase inhibitor staurosporine. Stimulation of apical transport exceeds stimulation of basolateral transport by up to 5-fold. We also find that the G protein activator aluminum fluoride, which stimulates transport to the surface at low fluoride concentrations as previously reported, inhibits transport at higher concentrations. In the nonpolarized fibroblast cell line CV-1, neither staurosporine nor aluminum fluoride stimulates transport to the cell surface. Our results suggest that the phosphorylation-dephosphorylation system, like the G protein, may be involved in the specialized sorting process characteristic of polarized cells. We show some evidence that these two mechanisms of regulation may act through common intermediates.
Brewer
C B
CB
Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas 75235-9038, USA.
Roth
M G
MG
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D016764:Cell Polarity; D002522:Chlorocebus aethiops; D004285:Dogs; D004988:Ethers, Cyclic / Q000494:pharmacology; D005089:Exocytosis; D019204:GTP-Binding Proteins / Q000378:metabolism; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D019311:Staurosporine
D000470:Alkaloids; D004988:Ethers, Cyclic; D047428:Protein Kinase Inhibitors; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D019204:GTP-Binding Proteins; D019311:Staurosporine