College of Life Science, Henan Agricultural University, Zhengzhou 450002, China.
Genes (Basel). 2021 Sep 30;12(10):1557. doi: 10.3390/genes12101557.
Pseudorabies virus (PRV) is recognized as one of the most important pathogens of swine and poses a serious threat to the swine industry worldwide. Available commercial vaccines fail to protect against the emergence of new PRV strains. Therefore, the new protein targets against PRV highlight the urgent need for uncovering the molecular determinants of host cellular proteins following PRV infection. Interferon-stimulated gene 15 (ISG15) demonstrates an outstanding antiviral response. However, the molecular mechanism of ISG15 that affects PRV replication is incompletely known. Here, we performed a tandem mass tag (TMT)-based approach to quantitatively identify protein expression changes in PRV-infected ISG15 knockout PK15 (ISG15-PK15) cells. In total, 4958 proteins were identified by using TMT coupled with LC-MS/MS in this study. In the PRV- and mock-infected groups, 241 differentially expressed proteins (DEPs) were identified, 162 upregulated and 79 downregulated proteins at 24 h post-infection (hpi), among which AFP, Vtn, Hsp40, Herc5, and Mccc1 may play important roles in PRV propagation. To ensure the validity and reliability of the proteomics data, the randomly selected DEPs were verified by RT-qPCR and Western blot analysis, and the results were consistent with the TMT results. Bioinformatics analyses further demonstrated that the DEPs are mainly involved in various biological processes and signaling pathways, such as signal transduction, the digestive system, and the PI3K-AKT pathway. These findings may provide new insight into molecular mechanisms for PRV infection, which is helpful for identifying potential protein targets for antiviral agents.
伪狂犬病病毒 (PRV) 被认为是猪的最重要病原体之一，对全球养猪业构成严重威胁。现有的商业疫苗无法预防新 PRV 毒株的出现。因此，针对 PRV 的新蛋白靶标突显了揭示 PRV 感染后宿主细胞蛋白分子决定因素的迫切需要。干扰素刺激基因 15 (ISG15) 表现出出色的抗病毒反应。然而，ISG15 影响 PRV 复制的分子机制尚不完全清楚。在这里，我们使用串联质量标签 (TMT) 方法定量鉴定 PRV 感染的 ISG15 敲除 PK15 (ISG15-PK15) 细胞中的蛋白质表达变化。在这项研究中，通过 TMT 与 LC-MS/MS 联用共鉴定到 4958 种蛋白质。在 PRV 和 mock 感染组中，在感染后 24 小时 (hpi) 鉴定到 241 个差异表达蛋白 (DEPs)，其中 162 个上调，79 个下调，AFP、Vtn、Hsp40、Herc5 和 Mccc1 可能在 PRV 繁殖中发挥重要作用。为了确保蛋白质组学数据的有效性和可靠性，通过 RT-qPCR 和 Western blot 分析随机选择的 DEPs 进行了验证，结果与 TMT 结果一致。生物信息学分析进一步表明，DEPs 主要参与各种生物过程和信号通路，如信号转导、消化系统和 PI3K-AKT 通路。这些发现可能为 PRV 感染的分子机制提供新的见解，有助于鉴定抗病毒药物的潜在蛋白靶标。
