Wilson Alexandra, Kamel Wael, Davies Kelsey, De Laurent Zaydah R, Arif Rozeena, Clarke Andrew T, Bell-Sakyi Lesley, Lamont Douglas, Demyanenko Yana, Noerenberg Marko, Kohl Alain, Mohammed Shabaz, Castello Alfredo, Brennan Benjamin
Medical Research Council-University of Glasgow Centre for Virus Research, Glasgow, Scotland, United Kingdom.
Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
PLoS Pathog. 2025 Aug 4;21(8):e1013393. doi: 10.1371/journal.ppat.1013393. eCollection 2025 Aug.
Cellular RNA-binding proteins (RBPs) are pivotal for the viral lifecycle, mediating key host-virus interactions that promote or repress virus infection. While these interactions have been largely studied in the vertebrate host, no comprehensive analyses of protein-RNA interactions occurring in cells of arbovirus vectors, in particular ticks, have been performed to date. Here we systematically identified the responses of the RNA-binding proteome (RBPome) to infection with a prototype bunyavirus (Uukuniemi virus; UUKV) in tick cells and discovered changes in RNA-binding activity for 283 proteins. In an orthogonal approach, we analysed the composition of the viral ribonucleoprotein by immunoprecipitation of UUKV nucleocapsid protein (N) in infected cells. We found many tick RBPs that are regulated by UUKV infection and associate with viral nucleocapsid protein complexes, and we confirmed experimentally that they impact UUKV infection. This includes the tick homolog of topoisomerase 3B (TOP3B), a protein able to manipulate the topology of RNA, which particularly affected viral particle production. Our data thus reveals the first protein-RNA interaction map for infected tick cells.
细胞RNA结合蛋白(RBPs)对病毒生命周期至关重要,介导促进或抑制病毒感染的关键宿主-病毒相互作用。虽然这些相互作用在脊椎动物宿主中已得到广泛研究,但迄今为止,尚未对虫媒病毒载体(特别是蜱虫)细胞中发生的蛋白质-RNA相互作用进行全面分析。在这里,我们系统地鉴定了蜱虫细胞中RNA结合蛋白质组(RBPome)对原型布尼亚病毒(乌库涅米病毒;UUKV)感染的反应,并发现283种蛋白质的RNA结合活性发生了变化。通过正交方法,我们通过免疫沉淀感染细胞中的UUKV核衣壳蛋白(N)来分析病毒核糖核蛋白的组成。我们发现许多蜱虫RBPs受UUKV感染调控并与病毒核衣壳蛋白复合物相关联,并且我们通过实验证实它们影响UUKV感染。这包括拓扑异构酶3B(TOP3B)的蜱虫同源物,一种能够操纵RNA拓扑结构的蛋白质,它对病毒粒子产生有特别影响。因此,我们的数据揭示了感染蜱虫细胞的首个蛋白质-RNA相互作用图谱。