Mutation of Ha-Ras C terminus changes effector pathway utilization

J Biol Chem. 2000 Aug 4;275(31):23559-68. doi: 10.1074/jbc.M001368200.

Abstract

In PC12 cells, Ha-Ras modulates multiple effector proteins that induce neuronal differentiation. To regulate these pathways Ha-Ras must be located at the plasma membrane, a process normally requiring attachment of farnesyl and palmitate lipids to the C terminus. Ext61L, a constitutively activated and palmitoylated Ha-Ras that lacks a farnesyl group, induced neurites with more actin cytoskeletal changes and lamellipodia than were induced by farnesylated Ha-Ras61L. Ext61L-triggered neurite outgrowth was prevented easily by co-expressing inhibitory Rho, Cdc42, or p21-activated kinase but required increased amounts of inhibitory Rac. Compared with Ha-Ras61L, Ext61L caused 2-fold greater Rac GTP binding and phosphatidylinositol 3-kinase activity in membranes, a hyperactivation that explained the numerous lamellipodia and ineffectiveness of Rac(N17). In contrast, Ext61L activated B-Raf kinase and ERK phosphorylation more poorly than Ha-Ras61L. Thus, accentuated differentiation by Ext61L apparently results from heightened activation of one Ras effector (phosphatidylinositol 3-kinase) and suboptimal activation of another (B-Raf). This surprising unbalanced effector activation, without changes in the designated Ras effector domain, indicates the Ext61L C-terminal alternations are a new way to influence Ha-Ras-effector utilization and suggest a broader role of the lipidated C terminus in Ha-Ras biological functions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actins
  • Animals
  • Cell Differentiation
  • Cytoskeleton
  • Models, Biological
  • Mutation
  • Neurites*
  • Neurons / cytology*
  • PC12 Cells
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Proto-Oncogene Proteins c-raf / metabolism*
  • Rats
  • Signal Transduction
  • ras Proteins / genetics
  • ras Proteins / metabolism*

Substances

  • Actins
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-raf
  • ras Proteins