Activation of G protein-coupled receptors (GPCRs) plays important roles in ovarian physiology. When GPCRs are bound by specific ligands, such as hormones, a series of signaling pathways can be initiated, leading to a variety of physiological events. GPCRs-mediated activation of protein kinase A (PKA) as well as protein kinase C (PKC) signaling pathways has been demonstrated to involve in regulation of proliferation, differentiation and cell survival in ovarian granulosa cells. The regulator of G protein signaling (RGS) is able to regulate GPCRs signaling by accelerating the deactivation of heterotrimeric G proteins via enhancing the intrinsic GTPase activity of Gα subunit, resulting in hydrolysis of GTP to GDP to cease the signaling. Currently the detailed mechanisms in PKA- and PKC-regulated RGS2 expression as well as translocation in ovarian system remain mostly unclear. In this study, we aimed to unveil the impacts of PKA and PKC activation in RGS2 expression and cellular localization in mouse granulosa cells. We first demonstrated that cAMP and phorbol-12, 13-didecanoate (PDD), a PKC activator were able to elevate both RGS2 protein and mRNA expression, as determined by western blotting and reverse transcriptase-PCR assays. Transient transfection of granulosa cells with the plasmids carrying a full-length or various sizes in length of mouse RGS2 promoter sequence into granulosa cells allowed us to observe that both PKA and PKC appeared to mediate RGS2 promoter activation and a short 78 bp fragment of this promoter sequence still retained a full-inducibility by cAMP and PDD. In addition, two consensus Sp1 sites within the 78 promoter sequence were mutated individually or in combination to identify that a Sp1 element was an essential component to regulate PKA- and PKC-induced promoter activation. Finally, we also monitored RGS2 protein expression/localization in total cell lysates, nuclear and cytosolic compartments and the results indicated an upregulation in total and in nuclear and cytosolic expression by cAMP and PDD. Similar results were also observed by transfecting a FLAG-RGS2 fusion plasmid into COS7 cells and we noted that PDD indeed induced a translocation of nuclear FLAG-RGS2 fusion protein to cytosol within 1 hr. Our findings suggest that expression of RGS2 protein and mRNA could be enhanced by PKA-dependent and PKC-dependent pathways with the Sp1 element within the RGS2 promoter sequence being the important component to mediate PKA- and PKC-induced RGS2 transcription.