Jansens Robert J J, Marmiroli Sandra, Favoreel Herman W
Department of Virology, Parasitology and Immunology, Faculty of Veterinary Medicine, Ghent University, 9820 Merelbeke, Belgium.
Cellular Signaling Laboratory, Department of Surgery, Medicine, Dentistry, and Morphology, University of Modena & Reggio Emilia, 41121 Modena, Italy.
Pathogens. 2020 Nov 5;9(11):916. doi: 10.3390/pathogens9110916.
The US3 serine/threonine protein kinase is conserved among the alphaherpesvirus family and represents an important virulence factor. US3 plays a role in viral nuclear egress, induces dramatic alterations of the cytoskeleton, represses apoptosis, enhances gene expression and modulates the immune response. Although several substrates of US3 have been identified, an unbiased screen to identify US3 phosphorylation targets has not yet been described. Here, we perform a shotgun and phosphoproteomics analysis of cells expressing the US3 protein of pseudorabies virus (PRV) to identify US3 phosphorylation targets in an unbiased way. We identified several cellular proteins that are differentially phosphorylated upon US3 expression and validated the phosphorylation of lamin A/C at serine 404, both in US3-transfected and PRV-infected cells. These results provide new insights into the signaling network of the US3 protein kinase and may serve as a basis for future research into the role of the US3 protein in the viral replication cycle.
US3丝氨酸/苏氨酸蛋白激酶在α疱疹病毒家族中保守，是一种重要的毒力因子。US3在病毒核出芽中起作用，诱导细胞骨架发生显著改变，抑制细胞凋亡，增强基因表达并调节免疫反应。尽管已鉴定出US3的几种底物，但尚未描述用于鉴定US3磷酸化靶点的无偏向性筛选方法。在此，我们对表达伪狂犬病病毒（PRV）US3蛋白的细胞进行鸟枪法和磷酸化蛋白质组学分析，以无偏向性方式鉴定US3磷酸化靶点。我们鉴定出几种在US3表达时发生差异磷酸化的细胞蛋白，并验证了在转染US3的细胞和感染PRV的细胞中，核纤层蛋白A/C在丝氨酸404处的磷酸化。这些结果为US3蛋白激酶的信号网络提供了新见解，并可能为未来研究US3蛋白在病毒复制周期中的作用奠定基础。
