College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, 712100, China; College of Agriculture and Animal Husbandry, Qinghai University, Xining, Qinghai, 810000, China.
College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi, 712100, China; School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, Henan, 453003, China.
Vet Microbiol. 2021 Sep;260:109093. doi: 10.1016/j.vetmic.2021.109093. Epub 2021 Jun 30.
The interactions between host cellular proteins and viral proteins are important for successful infection by viruses. Previous studies from our group have identified various host cellular proteins that can interact with the Newcastle disease virus V protein (Chu et al., 2018a), but their function in NDV replication has not been fully determined. The present study reports that heterogenous nuclear ribonucleoprotein H1 (hnRNP H1) can interact with NDV V protein in yeast. The immunofluorescence results showed that hnRNP H1 and V protein could colocalize in the cytoplasm of a chicken embryo fibroblast cell line (DF-1 cells). Co-immunoprecipitation assays further verified the interaction of these two proteins. The effects of overexpression and knockdown of hnRNP H1 on NDV replication were evaluated in DF-1 cells through real time quantitative PCR (RT-qPCR) and plaque assays. The regulation of V protein on hnRNP H1 expression was also examined. The results indicated that overexpression of hnRNP H1 facilitated NDV replication, while knockdown of hnRNP H1 decreased NDV replication. It was also shown that V protein could regulate hnRNP H1 expression at the protein level instead of the transcription level. The effect of V protein and hnRNP H1 on the DF-1 cell cycle was also tested and the results revealed that V protein may regulate cell proliferation by controlling the expression of hnRNP H1. Taken together, these results suggest that NDV V protein could promote viral replication by interacting with hnRNP H1.
宿主细胞蛋白与病毒蛋白之间的相互作用对病毒的成功感染很重要。我们小组的先前研究已经鉴定了各种宿主细胞蛋白,它们可以与新城疫病毒 V 蛋白相互作用(Chu 等人,2018a),但它们在 NDV 复制中的功能尚未完全确定。本研究报告称异质核核糖核蛋白 H1(hnRNP H1)可以与 NDV V 蛋白在酵母中相互作用。免疫荧光结果表明,hnRNP H1 和 V 蛋白可以在鸡胚成纤维细胞系(DF-1 细胞)的细胞质中共定位。共免疫沉淀实验进一步验证了这两种蛋白质的相互作用。通过实时定量 PCR(RT-qPCR)和蚀斑测定评估了 hnRNP H1 在 DF-1 细胞中对 NDV 复制的过表达和敲低的影响。还检查了 V 蛋白对 hnRNP H1 表达的调节作用。结果表明,hnRNP H1 的过表达促进了 NDV 的复制,而 hnRNP H1 的敲低则降低了 NDV 的复制。还表明 V 蛋白可以在蛋白质水平而不是转录水平上调节 hnRNP H1 的表达。还测试了 V 蛋白和 hnRNP H1 对 DF-1 细胞周期的影响,结果表明 V 蛋白可能通过控制 hnRNP H1 的表达来调节细胞增殖。总之,这些结果表明 NDV V 蛋白可以通过与 hnRNP H1 相互作用来促进病毒复制。