Tyrosine hydroxylase (TH) and dopa decarboxylase (DDC) are essential enzymes for dopamine (DA) biosynthesis. We found previously that V-1 overexpression elevates TH and DDC mRNA levels in culture (JBC 1998), raising the possibility of application of V-1 gene therapy for Parkinson's disease (PD). Here, we provide the first evidence for a novel regulatory mechanism of coordinated gene expression of TH/DDC/Nurr1 by V-1, an actin dynamics regulator. Knockdown of endogenous V-1 reduced TH, DDC and Nurr1 expression with reduction of SRF-mediated transcription in cultured mesencephalic neurons (DA neurons), which concomitantly decreased RhoA activity and actin assembly. Conversely, V-1 overexpression promoted RhoA activity and actin assembly to elevate TH/DDC/Nurr1 gene expression, accompanied with enhancement of MAL nuclear import and DA biosynthesis. ChIP-PCR assay with anti-SRF antibody identified the V-1 regulated SRF-binding in the promoter region of above three genes. Interestingly, VSV-G/V-1 transfection alleviated MPTP-induced neurodegeneration of DA neurons and recovered impaired expression levels of TH, DDC, Nurr1 and VMAT2 as well as decreased cofilin phosphorylation and SRF-mediated transcription. Injection of VSV-G/V-1 into the SNc of C57BL6 mice also elevated TH, DDC, Nurr1 and VMAT2 levels in nigrostriatum in vivo and augmented DA biosynthesis without an increase of L-DOPA, which caused no abnormal behavior. We conclude that V-1 facilitates RhoA-actin signaling, thereby inducing MAL nuclear accumulation, which activates the transcription of SRF target genes, including TH/DDC/Nurr1 gene in dopaminergic system, indicating V-1's potential benefit for safe and fundamental therapy of PD.