Difference cloning has identified a Villin-like mRNA transcript expressed selectively in peripheral sensory and sympathetic neurons. Pervin, the encoded 820-amino acid protein, has 60% identity with Villin and is the rat homologue of Advillin. Coimmunoprecipitation studies demonstrate that Pervin and actin interact in vivo. Transfection of COS-7 epithelial cell lines demonstrates colocalization of epitope-tagged Pervin with green fluorescent protein-actin and results in an increase in process formation. This effect is abolished when the putative actin-bundling headpiece of Pervin is deleted. Biolistic transfection of primary cultures of rat dorsal root ganglion sensory neurons also results in increased neurite outgrowth with FLAG-tagged Pervin. Deletion of the actin-bundling headpiece inhibits normal neurite growth. These data suggest that Pervin may play a significant role in regulating process outgrowth in peripheral neurons through a mechanism that involves the activity of an actin-bundling domain.