Ras pathway activates epithelial Na+ channel and decreases its surface expression in Xenopus oocytes

Mol Biol Cell. 1998 Dec;9(12):3417-27. doi: 10.1091/mbc.9.12.3417.

Abstract

The small G protein K-Ras2A is rapidly induced by aldosterone in A6 epithelia. In these Xenopus sodium reabsorbing cells, aldosterone rapidly activates preexisting epithelial Na+ channels (XENaC) via a transcriptionally mediated mechanism. In the Xenopus oocytes expression system, we tested whether the K-Ras2A pathway impacts on XENaC activity by expressing XENaC alone or together with XK-Ras2A rendered constitutively active (XK-Ras2AG12V). As a second control, XENaC-expressing oocytes were treated with progesterone, a sex steroid that induces maturation of the oocytes similarly to activated Ras. Progesterone or XK-Ras2AG12V led to oocyte maturation characterized by a decrease in surface area and endogenous Na+ pump function. In both conditions, the surface expression of exogenous XENaC's was also decreased; however, in comparison with progesterone-treated oocytes, XK-ras2AG12V-coinjected oocytes expressed a fivefold higher XENaC-mediated macroscopic Na+ current that was as high as that of control oocytes. Thus, the Na+ current per surface-expressed XENaC was increased by XK-Ras2AG12V. The chemical driving force for Na+ influx was not changed, suggesting that XK-Ras2AG12V increased the mean activity of XENaCs at the oocyte surface. These observations raise the possibility that XK-Ras2A, which is the first regulatory protein known to be transcriptionally induced by aldosterone, could play a role in the control of XENaC function in aldosterone target cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldosterone / pharmacology
  • Animals
  • Cell Line
  • Cell Membrane / metabolism
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Female
  • Gene Expression
  • In Vitro Techniques
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Progesterone / pharmacology
  • Sodium Channels / drug effects
  • Sodium Channels / genetics
  • Sodium Channels / metabolism*
  • Xenopus
  • ras Proteins / genetics
  • ras Proteins / metabolism*

Substances

  • Sodium Channels
  • Aldosterone
  • Progesterone
  • ras Proteins