Treffers Emmely E, Tas Ali, Scholte Florine E M, Van Myrthe N, Heemskerk Matthias T, de Ru Arnoud H, Snijder Eric J, van Hemert Martijn J, van Veelen Peter A
Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, ZA, Leiden, The Netherlands.
Department of Immunohematology and Blood transfusion, Leiden University Medical Center, ZA, Leiden, The Netherlands.
Proteomics. 2015 Jul;15(13):2267-80. doi: 10.1002/pmic.201400581. Epub 2015 Apr 22.
Chikungunya virus (CHIKV) is an arthropod-borne reemerging human pathogen that generally causes a severe persisting arthritis. Since 2005, the virus has infected millions of people during outbreaks in Africa, Indian Ocean Islands, Asia, and South/Central America. Many steps of the replication and expression of CHIKV's 12-kb RNA genome are highly dependent on cellular factors, which thus constitute potential therapeutic targets. SILAC and LC-MS/MS were used to define the temporal dynamics of the cellular response to infection. Using samples harvested at 8, 10, and 12 h postinfection, over 4700 proteins were identified and per time point 2800-3500 proteins could be quantified in both biological replicates. At 8, 10, and 12 h postinfection, 13, 38, and 106 proteins, respectively, were differentially expressed. The majority of these proteins showed decreased abundance. Most subunits of the RNA polymerase II complex were progressively degraded, which likely contributes to the transcriptional host shut-off observed during CHIKV infection. Overexpression of four proteins that were significantly downregulated (Rho family GTPase 3 (Rnd3), DEAD box helicase 56 (DDX56), polo-like kinase 1 (Plk1), and ubiquitin-conjugating enzyme E2C (UbcH10) reduced susceptibility of cells to CHIKV infection, suggesting that infection-induced downregulation of these proteins is beneficial for CHIKV replication. All MS data have been deposited in the ProteomeXchange with identifier PXD001330 (http://proteomecentral.proteomexchange.org/dataset/PXD001330).
基孔肯雅病毒（CHIKV）是一种节肢动物传播的再度出现的人类病原体，通常会导致严重的持续性关节炎。自2005年以来，该病毒在非洲、印度洋岛屿、亚洲以及南美洲/中美洲的疫情爆发期间感染了数百万人。CHIKV 12 kb RNA基因组的复制和表达的许多步骤高度依赖细胞因子，因此这些细胞因子构成了潜在的治疗靶点。采用稳定同位素标记氨基酸法（SILAC）和液相色谱-串联质谱法（LC-MS/MS）来确定细胞对感染反应的时间动态。利用感染后8、10和12小时采集的样本，鉴定出了超过4700种蛋白质，并且在每个时间点，两个生物学重复中均可对2800 - 3500种蛋白质进行定量分析。在感染后8、10和12小时，分别有13、38和106种蛋白质差异表达。这些蛋白质中的大多数丰度降低。RNA聚合酶II复合物的大多数亚基逐渐降解，这可能导致了基孔肯雅病毒感染期间观察到的转录宿主关闭现象。四种显著下调的蛋白质（Rho家族GTP酶3（Rnd3）、DEAD盒解旋酶56（DDX56）、polo样激酶1（Plk1）和泛素结合酶E2C（UbcH10））的过表达降低了细胞对基孔肯雅病毒感染的易感性，这表明感染诱导的这些蛋白质下调有利于基孔肯雅病毒的复制。所有质谱数据已存入蛋白质组交换库，标识符为PXD001330（http://proteomecentral.proteomexchange.org/dataset/PXD001330）。
