Rho regulates cytoskeletal actin structure and integrin-mediated cell adhesion. Rho also has a role in cell growth regulation and is required for cell transformation by oncogenic Ras. Recently, it has been demonstrated that Rho can activate transcription from the c-fos serum response element (SRE). This raised the possibility that functions required for Rho-mediated cell transformation might overlap with those involved in transcriptional regulation. Here we show that RhoB can activate the SRE and can synergize in cell transformation with constitutively activated Raf-CAAX. Significantly, unprenylated forms of RhoB that are biologically inert and unable to transform cells can still activate SRE-dependent transcription. This finding suggests that transcriptional activation by Rho may be separable from its cell transforming functions.