The budded form of Autographa californica M nucleopolyhedrovirus enters permissive cells via adsorptive endocytosis. Shortly after nucleocapsid penetration into the cytoplasm, thick actin cables form, which frequently project toward the nucleus. These actin cables are transient structures, formed in association with viral nucleocapsids prior to viral gene expression and concomitant with nucleocapsid transport to the nucleus. In this paper we report that nucleocapsids are capable of nucleating actin polymerization in vitro in a concentration-dependent manner. Two viral-encoded capsid proteins, p39 and p78/83, were found to bind actin directly and therefore could be involved in the observed acceleration of actin polymerization. When nucleocapsids were added to actin in the presence of cytochalasin D, actin polymerization was reduced to levels below those obtained with actin and cytochalasin D alone, suggesting that the nucleocapsids bound to the pointed ends of actin filaments. Finally, treatment of infected cells with the myosin inhibitor 2,3-butanedione monoxime delayed nucleocapsid transport to the nucleus. We postulate that upon entering the cytoplasm, AcMNPV nucleocapsids induce the polymerization of actin cables, which, in conjunction with a myosin-like motor, facilitate their transport to and/or into the nucleus.