Instituto de Biología Integrativa de Sistemas (I2SysBio), CSIC - Universitat de València, 46980, Paterna, Spain.
Instituto de Biología Molecular y Celular de Plantas (IBMCP), CSIC - Universitat Politècnica de València, 46022, València, Spain.
Commun Biol. 2023 Jan 11;6(1):28. doi: 10.1038/s42003-023-04427-8.
Viruses are obligate intracellular parasites that have co-evolved with their hosts to establish an intricate network of protein-protein interactions. Here, we followed a high-throughput yeast two-hybrid screening to identify 378 novel protein-protein interactions between turnip mosaic virus (TuMV) and its natural host Arabidopsis thaliana. We identified the RNA-dependent RNA polymerase NIb as the viral protein with the largest number of contacts, including key salicylic acid-dependent transcription regulators. We verified a subset of 25 interactions in planta by bimolecular fluorescence complementation assays. We then constructed and analyzed a network comprising 399 TuMV-A. thaliana interactions together with intravirus and intrahost connections. In particular, we found that the host proteins targeted by TuMV are enriched in different aspects of plant responses to infections, are more connected and have an increased capacity to spread information throughout the cell proteome, display higher expression levels, and have been subject to stronger purifying selection than expected by chance. The proviral or antiviral role of ten host proteins was validated by characterizing the infection dynamics in the corresponding mutant plants, supporting a proviral role for the transcriptional regulator TGA1. Comparison with similar studies with animal viruses, highlights shared fundamental features in their mode of action.
病毒是专性细胞内寄生虫,它们与宿主共同进化,形成了一个复杂的蛋白质-蛋白质相互作用网络。在这里,我们通过高通量酵母双杂交筛选,鉴定了芜菁花叶病毒(TuMV)与其天然宿主拟南芥之间的 378 种新的蛋白质-蛋白质相互作用。我们发现 RNA 依赖性 RNA 聚合酶 NIb 是与宿主互作最多的病毒蛋白,其中包括关键的水杨酸依赖型转录调控因子。我们通过双分子荧光互补实验验证了 25 个互作中的一部分。然后,我们构建并分析了一个包含 399 个 TuMV-拟南芥相互作用以及病毒内和宿主内连接的网络。特别是,我们发现 TuMV 靶向的宿主蛋白在植物对感染的反应的不同方面富集,它们之间的连接更多,并且具有在整个细胞蛋白质组中传播信息的更高能力,表达水平更高,并且受到的净化选择压力比预期的随机选择更大。通过对相应突变体植物的感染动力学进行表征,验证了十个宿主蛋白的助病毒或抗病毒作用,支持转录调控因子 TGA1 具有助病毒作用。与动物病毒的类似研究进行比较,突出了它们作用模式的共同基本特征。