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Crimean‐Congo hemorrhagic fever virus is the causative agent of a severe disease throughout Africa, Europe, and Asia. Like other members of the genus Nairovirus, the Crimean‐Congo hemorrhagic fever virus contains three genomic RNA segments, the small (S), medium (M), and large (L) segments. The S segment encodes the viral nucleocapsid protein (NP), while the M and L segments encode the glycoproteins and the RNA‐dependent RNA polymerase, respectively. In this study, the site of expression and assembly of Crimean‐Congo hemorrhagic fever virus NP in mammalian cells have been investigated. It was found that the NP is localized in the perinuclear region of infected cells. By using the Semliki forest virus expression system, it was shown that the Crimean‐Congo hemorrhagic fever virus NP is targeted to the perinuclear region of cells in the absence of native RNA segments and virally encoded glycoproteins. It was also demonstrated that the Crimean‐Congo hemorrhagic fever virus NP was not expressed as a Golgi‐membrane associated protein. By using Cytochalasin D, an agent that disrupts actin filaments, it was found that actin filaments are involved in targeting the viral NP to perinuclear regions. We also demonstrated that disruption of actin filaments reduced the assembly of infectious Crimean‐Congo hemorrhagic fever virus up to 97%. Furthermore, we showed that the NP of Crimean‐Congo hemorrhagic fever virus NP interacts with actin. J. Med. Virol. 72:83–93, 2004. © 2004 Wiley‐Liss, Inc.