Andersson Ida, Simon Melinda, Lundkvist Ake, Nilsson Mikael, Holmström Anna, Elgh Fredrik, Mirazimi Ali
Center for Microbiological Preparedness,Swedish Institute for Infectious Disease Control, SE Solna, Sweden.
J Med Virol. 2004 Jan;72(1):83-93. doi: 10.1002/jmv.10560.
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.
克里米亚-刚果出血热病毒是非洲、欧洲和亚洲一种严重疾病的病原体。与内罗病毒属的其他成员一样，克里米亚-刚果出血热病毒包含三个基因组RNA片段，即小（S）、中（M）和大（L）片段。S片段编码病毒核衣壳蛋白（NP），而M和L片段分别编码糖蛋白和RNA依赖性RNA聚合酶。在本研究中，对克里米亚-刚果出血热病毒NP在哺乳动物细胞中的表达和组装位点进行了研究。发现NP定位于受感染细胞的核周区域。通过使用辛德毕斯病毒表达系统，表明在没有天然RNA片段和病毒编码糖蛋白的情况下，克里米亚-刚果出血热病毒NP靶向细胞的核周区域。还证明克里米亚-刚果出血热病毒NP不是作为高尔基体膜相关蛋白表达的。通过使用细胞松弛素D（一种破坏肌动蛋白丝的试剂），发现肌动蛋白丝参与将病毒NP靶向核周区域。我们还证明，肌动蛋白丝的破坏使传染性克里米亚-刚果出血热病毒的组装减少了97%。此外，我们表明克里米亚-刚果出血热病毒NP与肌动蛋白相互作用。
