Successful utilization of myogenic progenitor cells for therapeutic applications requires an understanding of the intrinsic and extrinsic cues involved in their regulation. Here we demonstrate the expression pattern and transcriptional regulation of Rad, a prototypic member of a family of novel Ras-related GTPases, during mammalian development and skeletal muscle regeneration. Rad was identified from microarray analysis based on the robust upregulation of its expression during skeletal muscle regeneration. Our current findings demonstrate negligible Rad expression within resting adult skeletal muscle, however following muscle injury, Rad expression is significantly increased and appears to be localized to the myogenic progenitor cell population during the early phases of regeneration and within the newly regenerated myofibers during the later phases of regeneration. This expression profile of Rad during skeletal muscle regeneration is consistent with the proposed roles of Rad in the inhibition of L-type Ca²⁺ channel activity and the inhibition of Rho/ROK activity. We also demonstrate that known myogenic transcription factors (MEF2, MyoD and Myf-5) can increase the transcriptional activity of the Rad promoter and this ability is significantly enhanced by the presence of the calcium-dependent phosphatase, calcineurin. Furthermore, this enhanced transcriptional activity appears to be dependent on the presence of a conserved NFAT binding motif within the Rad promoter. Taken together, these data define Rad as a novel factor within the regenerative process of skeletal muscle and identify key regulators of its transcriptional activity.