Aldosterone induces ras methylation in A6 epithelia

• Published in: American Journal of Physiology: Cell Physiology Vol. 279; no. 2; p. C429
• Main Authors: Al-Baldawi, N F, Stockand, J D, Al-Khalili, O K, Yue, G, Eaton, D C
• Format: Journal Article
• Language: English
• Published: United States 01-08-2000
• Subjects: Aldosterone - pharmacology; Animals; Cells, Cultured; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Kidney Tubules - drug effects; Kidney Tubules - metabolism; Methylation; Protein Methyltransferases - drug effects; Protein Methyltransferases - metabolism; ras Proteins - drug effects; ras Proteins - metabolism; Sodium Channels - drug effects; Sodium Channels - metabolism
• ISSN: 0363-6143
• DOI: 10.1152/ajpcell.2000.279.2.c429

Aldosterone increases Na(+) reabsorption by renal epithelial cells: the acute actions (<4 h) appear to be promoted by protein methylation. This paper describes the relationship between protein methylation and aldosterone's action and describes aldosterone-mediated targets for methylation in cultured renal cells (A6). Aldosterone increases protein methylation from 7.90 +/- 0.60 to 20.1 +/- 0.80 methyl ester cpm/microg protein. Aldosterone stimulates protein methylation by increasing methyltransferase activity from 14.0 +/- 0.64 in aldosterone-depleted cells to 31.8 +/- 2.60 methyl ester cpm/microg protein per hour in aldosterone-treated cells. Three known methyltransferase inhibitors reduce the aldosterone-induced increase in methyltransferase activity. One of these inhibitors, the isoprenyl-cysteine methyltransferase-specific inhibitor, S-trans, trans-farnesylthiosalicylic acid, completely blocks aldosterone-induced protein methylation and also aldosterone-induced short-circuit current. Aldosterone induces protein methylation in two molecular weight ranges: near 90 kDa and around 20 kDa. The lower molecular weight range is the weight of small G proteins, and aldosterone does increase both Ras protein 1.6-fold and Ras methylation almost 12-fold. Also, Ras antisense oligonucleotides reduce the activity of Na(+) channels by about fivefold. We conclude that 1) protein methylation is essential for aldosterone-induced increases in Na(+) transport; 2) one target for methylation is p21(ras); and 3) inhibition of Ras expression or Ras methylation inhibits Na(+) channel activity.