Regulation of serum response factor (SRF)-mediated gene transcription by G protein subunits and G protein-coupled receptors was investigated in transfected NIH3T3 cells and in a cell line that was derived from mice lacking Galphaq and Galpha11. We found that the constitutively active forms of the alpha subunits of the Gq and G12 class of G proteins, including Galphaq, Galpha11, Galpha14, Galpha16, Galpha12, and Galpha13, can activate SRF in NIH3T3 cells. We also found that the type 1 muscarinic receptor (m1R) and alpha1-adrenergic receptor (AR)-mediated SRF activation is exclusively dependent on Galphaq/11, while the receptors for thrombin, lysophosphatidic acid (LPA), thromboxane A2, and endothelin can activate SRF in the absence of Galphaq/11. Moreover, RGS12 but not RGS2, RGS4, or Axin was able to inhibit Galpha12 and Galpha13-mediated SRF activation. And RGS12, but not other RGS proteins, blocked thrombin- and LPA-mediated SRF activation in the Galphaq/11-deficient cells. Therefore, the thrombin, LPA, thromboxane A2, and endothelin receptors may be able to couple to Galpha12/13. On the contrary, receptors including beta2- and alpha2-ARs, m2R, the dopamine receptors type 1 and 2, angiotensin receptors types 1 and 2, and interleukin-8 receptor could not activate SRF in the presence or absence of Galphaq/11, suggesting that these receptors cannot couple to endogenous G proteins of the G12 or Gq classes.