Involvement of Rho p21 small GTP-binding protein and its regulator in the HGF-induced cell motility

Oncogene. 1994 Jan;9(1):273-9.

Abstract

Hepatocyte growth factor (HGF) induced motility of cultured mouse keratinocytes (308R cells). This HGF-induced cell motility was inhibited by microinjection of either rho GDI, an inhibitory GDP/GTP exchange protein for rho p21 small GTP-binding protein, or a botulinum exoenzyme C3 which is known to selectively impair the function of rho p21 by ADP-ribosylating its effector domain. The rho GDI action was prevented by comicroinjection with the guanosine 5'-(3-0-thio)triphosphate (GTP gamma S)-bound active form of rhoA p21, and the C3 action was prevented by comicroinjection with a rhoA p21 mutant (rhoAIle41 p21) which is resistant to the C3 action. The HGF-induced cell motility was not inhibited by microinjection of a dominant negative rac1 p21 mutant (rac1Asn17 p21) or a dominant negative Ki-ras p21 mutant (Ki-rasAsn17 p21). Microinjection of the GTP gamma S-bound form of rac1 p21 or a dominant active Ki-ras p21 mutant (Ki-rasVal12 p21) did not induce cell motility. These results indicate that both rho p21 and rho GDI, but neither rac p21 nor ras p21, are involved in the HGF-induced cell motility. However, microinjection of the GTP gamma S-bound form of rhoA p21 alone did not induce cell motility in the absence of HGF, suggesting that activation of rho p21 is necessary but not sufficient for the HGF-induced cell motility. The HGF-induced cell motility was mimicked by 12-0-tetradecanoyl-phorbol-13-acetate, a protein kinase C-activating phorbol ester, but not by Ca2+ ionophore. The phorbol ester-induced cell motility was also inhibited by microinjection of rho GDI or C3. These results indicate that both rho p21 and rho GDI are also involved in the phorbol ester-induced cell motility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / physiology
  • Animals
  • Botulinum Toxins*
  • Cell Movement / drug effects
  • GTP-Binding Proteins / physiology*
  • Hepatocyte Growth Factor / pharmacology*
  • Mice
  • Microinjections
  • Protein Kinase C / physiology
  • Proto-Oncogene Proteins c-met
  • Proto-Oncogene Proteins p21(ras) / physiology
  • Receptor Protein-Tyrosine Kinases / physiology
  • Tetradecanoylphorbol Acetate / pharmacology
  • rhoA GTP-Binding Protein

Substances

  • Hepatocyte Growth Factor
  • ADP Ribose Transferases
  • exoenzyme C3, Clostridium botulinum
  • Proto-Oncogene Proteins c-met
  • Receptor Protein-Tyrosine Kinases
  • Protein Kinase C
  • Botulinum Toxins
  • GTP-Binding Proteins
  • Proto-Oncogene Proteins p21(ras)
  • rhoA GTP-Binding Protein
  • Tetradecanoylphorbol Acetate