PKC and Pi deprivation modulate differently the ubiquitous Na-dependent Pi uptake in MDCK cells.
The role of protein kinase C (PKC) in the modulation of the ubiquitous sodium-dependent phosphate transport and in adaptation of that transport to phosphate deprivation was investigated in MDCK cells. Phorbol myristate acetate (PMA) had a biphasic effect on sodium-dependent phosphate uptake characterized by early inhibition (-25% at 1 h) followed by late stimulation (2.3-fold at 15 h). Late stimulation was related to a decreased apparent affinity (Km) with unchanged maximal velocity (Vmax). The 15-h stimulation of phosphate uptake was also induced by an initial 1-h PMA treatment followed by a 14-h washout of PMA or by R59 022. The stimulation was inhibited by PKC downregulation. PMA stimulation was dependent on protein synthesis but not on transcription, as shown by the respective effects of cycloheximide, 3'-deoxyadenosine, and actinomycin D. In phosphate-deprived cells PMA had also a biphasic effect. A potentiation of PMA stimulation of phosphate uptake with phosphate deprivation was observed. Adaptation to phosphate deprivation was not prevented by PKC downregulation. Cytosolic and membranous PKC activities were not changed by 15-h phosphate deprivation. We conclude that 1) PKC modulates sodium-dependent phosphate uptake in MDCK cells, and 2) phosphate deprivation and PKC modulation of sodium-dependent phosphate uptake involve different cellular pathways; that is, phosphate deprivation acts through gene regulation, and PKC acts through translation regulation.
Escoubet
B
B
Department of Physiology, Faculté de Médecine X. Bichat, Institut National de la Santé et de la Recherche Médicale Unité 251, Paris, France.
Garestier
M C
MC
Cherqui
G
G
Amiel
C
C
D000222:Adaptation, Physiological; D000818:Animals; D002460:Cell Line; D004075:Diglycerides / Q000378:metabolism; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism*; D010710:Phosphates / Q000172:deficiency / Q000378:metabolism*; D011493:Protein Kinase C / Q000502:physiology*; D011744:Pyrimidinones / Q000494:pharmacology; D012964:Sodium / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology
D004075:Diglycerides; D010710:Phosphates; D011744:Pyrimidinones; D013844:Thiazoles; C047505:R 59022; D012964:Sodium; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate