Identification of amino acid residues of Ras protein that are essential for signal-transducing activity but not for enhancement of GTPase activity by GAP

FEBS Lett. 1991 Dec 9;294(3):187-90. doi: 10.1016/0014-5793(91)80665-p.

Abstract

To determine the amino acid residues required for the signal-transducing activity of the human c-Ha-Ras protein, we introduced point mutations at residues 45-54 near the 'effector region' (residues 32-40). We transfected PC12 cells with these mutant genes and also micro-injected the mutant proteins, bound with an unhydrolyzable GTP analog, into PC12 cells. Both procedures showed that Val45----Glu and Gly48----Cys mutations impaired the ability of the Ras protein to induce morphological change of PC12 cells. These mutations did not affect the guanine nucleotide-binding activity or GTPase activity in the absence or presence of bovine GTPase-activating protein (GAP). Therefore, the Val45 and Gly48 residues should be included by definition in the effector region responsible for the signal transduction, while only a subset of the effector-region residues is required for enhancement of the GTPase activity by GAP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Gland Neoplasms
  • Amino Acids / chemistry*
  • GTP Phosphohydrolases / metabolism*
  • GTPase-Activating Proteins
  • Genes, ras
  • Guanosine Diphosphate / metabolism
  • Guanosine Triphosphate / metabolism
  • Mutagenesis, Site-Directed
  • Pheochromocytoma
  • Proteins / pharmacology*
  • Proto-Oncogene Proteins p21(ras) / chemistry*
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Proto-Oncogene Proteins p21(ras) / metabolism
  • Signal Transduction*
  • Structure-Activity Relationship
  • Transfection
  • Tumor Cells, Cultured
  • ras GTPase-Activating Proteins

Substances

  • Amino Acids
  • GTPase-Activating Proteins
  • Proteins
  • ras GTPase-Activating Proteins
  • Guanosine Diphosphate
  • Guanosine Triphosphate
  • GTP Phosphohydrolases
  • Proto-Oncogene Proteins p21(ras)