鉴定和筛选与 ORFV-ORF047 蛋白相互作用的宿主蛋白。
Identification and screening of host proteins interacting with ORFV-ORF047 protein.
机构信息
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Public Health of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China.
School of Public Health, Lanzhou University, Lanzhou, 730046, China.
出版信息
Virol J. 2021 Jan 26;18(1):27. doi: 10.1186/s12985-021-01499-y.
BACKGROUND
Orf virus (ORFV) is a member of the genus Parapoxvirus and family Poxviridae. The virus has a worldwide distribution and infects sheep, goats, humans, and wild animals. However, due to the complex structure of the poxvirus, the underlying mechanism of the entry and infection by ORFV remains largely unknown. ORFV ORF047 encodes a protein named L1R. Poxviral L1R serves as the receptor-binding protein and blocks virus binding and entry independently of glycosaminoglycans (GAGs). The study aimed to identify the host interaction partners of ORFV ORF047.
METHODS
Yeast two-hybrid cDNA library of sheep testicular cells was applied to screen the host targets with ORF047 as the bait. ORF047 was cloned into a pBT3-N vector and expressed in the NMY51 yeast strain. Then, the expression of bait proteins was validated by Western blot analysis.
RESULTS
Sheep SERP1and PABPC4 were identified as host target proteins of ORFV ORF047, and a Co-IP assay further verified their interaction.
CONCLUSIONS
New host cell proteins SERP1and PABPC4 were found to interact with ORFV ORF047 and might involve viral mRNA translation and replication.
背景
口疮病毒（ORFV）是副痘病毒属和痘病毒科的成员。该病毒分布广泛，可感染绵羊、山羊、人类和野生动物。然而，由于痘病毒的结构复杂，ORFV 的进入和感染的潜在机制在很大程度上仍不清楚。ORFV ORF047 编码一种名为 L1R 的蛋白质。痘病毒的 L1R 作为受体结合蛋白，独立于糖胺聚糖（GAGs）阻断病毒结合和进入。本研究旨在鉴定 ORFV ORF047 的宿主相互作用伙伴。
方法
应用绵羊睾丸细胞的酵母双杂交 cDNA 文库筛选以 ORF047 为诱饵的宿主靶标。将 ORF047 克隆到 pBT3-N 载体中，并在 NMY51 酵母菌株中表达。然后，通过 Western blot 分析验证诱饵蛋白的表达。
结果
鉴定出绵羊 SERP1 和 PABPC4 为 ORFV ORF047 的宿主靶蛋白，Co-IP 实验进一步验证了它们的相互作用。
结论
发现新的宿主细胞蛋白 SERP1 和 PABPC4 与 ORFV ORF047 相互作用，可能参与病毒 mRNA 的翻译和复制。
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