Separate roles for H-Ras and Rac in signaling by transforming growth factor (TGF)-beta. H-Ras is essential for activation of MAP kinase, partially required for transcriptional activation by TGF-beta, but not required for signaling of growth suppression by TGF-beta.

The signaling components located downstream of the transforming growth factor (TGF)-beta receptor are poorly understood. We constructed adenoviral vectors expressing a dominant-negative form of either H-Ras (AdCARasY57) or Rac (AdCARacN17), and used them to examine the roles of H-Ras and Rac in TGF-beta signaling using arterial endothelial cells in primary culture, and several established cells including a mink lung epithelial cell line (Mv1Lu). The rapid activation of p42/44 MAP kinase (MAPK) by TGF-beta1 was eliminated completely, and transcriptional activation by TGF-beta1 of the plasminogen activator inhibitor-1 gene was reduced by 50% in both endothelial cells and Mv1Lu when they were infected with AdCARasY57. However, the antiproliferative effect of TGF-beta, as assessed by the induction of the mRNA for Cdk4/6-specific inhibitor p15INK4B and by DNA synthesis, was not affected in AdCARasY57-infected cells. A MAPK kinase (MEK)1/2 inhibitor, U0126 also abolished MAPK activation and partially inhibited transcriptional activation by TGF-beta, suggesting that MAPK may be partially involved in this pathway. MAPK activation, transcriptional activation and growth suppression by TGF-beta were all unaffected in cells infected with AdCARacN17, although the DNA synthesis elicited by serum mitogens was suppressed completely in the infected cells. Our data indicate that H-Ras is essential for mitogen-activated protein kinase activation, partly required for transcriptional activation by TGF-beta, but not critically involved in the signaling that exerts the antiproliferative effect of TGF-beta. The results also suggest that Rac may not serve as an essential molecule in signaling by TGF-beta in the cells tested.