Aldosterone rapidly activates protein kinase D via a mineralocorticoid receptor/EGFR trans-activation pathway in the M1 kidney CCD cell line.
Aldosterone elicits physiological responses through the modulation of gene expression and by stimulating signaling processes. Here we investigated the activation pathway of protein kinase D1 (PKD1) by aldosterone in the murine M1 renal cortical collecting duct cell line. Aldosterone stimulated a rapid increase in PKD1 activity peaking at 2-5 min and at 30 min after treatment that was insensitive to inhibitors of transcription or translation. PKD1 was not activated by aldosterone in MR null NIH-3T3 fibroblasts or M1-CCD cells propagated without dexamethasone, which did not express MR. PKD1 activation was sensitive to the MR antagonists spironolactone and RU28318 but not to the glucocorticoid receptor antagonist RU486. Aldosterone activation of PKD1 was inhibited by the epidermal growth factor (EGFR) antagonist tyrphostin AG1478 and by the c-Src inhibitor PP2. Western blotting revealed EGFR phosphorylation following aldosterone treatment at the c-Src tyrosine kinase-specific residue Tyr845. The activation of c-Src was dependent on its interaction with HSP84, since HSP84 antagonist 17-AAG inhibited both the phosphorylation of EGFR in response to aldosterone by c-Src and also the subsequent activation of PKD1.
Department of Molecular Medicine, Royal College of Surgeons in Ireland Education and Research Centre, Smurfit Building, Beaumont Hospital, Dublin 9, Ireland.
Victoria
V
McEneaney
Brian J
BJ
Harvey
Warren
W
Thomas
D000450:Aldosterone / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000235:genetics*; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D000451:Mineralocorticoid Receptor Antagonists; D041681:NIH 3T3 Cells; D011493:Protein Kinase C / Q000378:metabolism*; D011799:Quinazolines; D018161:Receptors, Mineralocorticoid / Q000235:genetics*; D013148:Spironolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D015533:Transcriptional Activation / Q000502:physiology*; D020032:Tyrphostins / Q000494:pharmacology
D000451:Mineralocorticoid Receptor Antagonists; D011799:Quinazolines; D018161:Receptors, Mineralocorticoid; D020032:Tyrphostins; C101044:RTKI cpd; D013148:Spironolactone; D000450:Aldosterone; C041006:RU 28318; C089212:protein kinase D; D066246:ErbB Receptors; D011493:Protein Kinase C