Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China; State Key Laboratory of Animal Nutrition, Beijing, China.
J Proteomics. 2019 Apr 15;197:34-41. doi: 10.1016/j.jprot.2019.02.011. Epub 2019 Feb 18.
H5N1 influenza A virus (IAV) causes seasonal epidemics that represent a worldwide threat to public health. IAV relies on host factors for viral replication. PB1 N40 is translated from the fifth starting code (AUG) of PB1 mRNA, which is the product of the ribosomal scan omission. Here, we report the interactome landscape of H5N1 IAV PB1 N40 protein in chicken cells. The interacting complexes were captured by co-immunoprecipitation and analyzed by mass spectrometry. We identified 135 proteins as PB1 N40-interacting proteins. GO and Pathway analysis showed that proteins with biological functions such as protein localization and viral transcription and proteins related to signaling pathways of DNA replication and cell cycle were significantly enriched in virus-host interactions, suggesting the potential roles of them in infection with H5N1 IAV. Comparative analysis among H1N1 and H5N1 revealed conservation of the virus-host protein interaction between different subtypes or strains of influenza virus. ARCN1 was identified as a host interacting factor of H5N1 IAV PB1 N40 protein, which is the component of the coatomer. Knockdown of ARCN1 significantly decreased the titer of H5N1 IAV in chicken cells. BIOLOGICAL SIGNIFICANCE: Influenza A virus (IAV) is a great threat to public health and avian production. However, the manner in which avian IAV recruits the host cellular machinery for replication and how the host antagonizes the IAV infection was previously poorly understood. Here we present the viral-host interactome of the H5N1 IAV PB1 N40 protein and reveal its involvement with dozens of important host genes during the course of IAV infection.
H5N1 流感病毒 (IAV) 引起季节性流行，对全球公共卫生构成威胁。IAV 依靠宿主因素进行病毒复制。PB1 N40 是从 PB1 mRNA 的第五个起始密码子 (AUG) 翻译而来的，这是核糖体扫描缺失的产物。在这里，我们报告了 H5N1 IAV PB1 N40 蛋白在鸡细胞中的互作组图谱。通过共免疫沉淀捕获相互作用复合物，并通过质谱分析进行分析。我们鉴定出 135 种蛋白质为 PB1 N40 相互作用蛋白。GO 和通路分析表明，具有蛋白质定位和病毒转录等生物学功能的蛋白质以及与 DNA 复制和细胞周期信号通路相关的蛋白质在病毒-宿主相互作用中显著富集，表明它们在感染 H5N1 IAV 中的潜在作用。在 H1N1 和 H5N1 之间进行比较分析表明，不同亚型或株流感病毒之间存在病毒-宿主蛋白相互作用的保守性。ARCN1 被鉴定为 H5N1 IAV PB1 N40 蛋白的宿主相互作用因子，是衣壳的组成部分。ARCN1 的敲低显著降低了 H5N1 IAV 在鸡细胞中的滴度。生物学意义：流感病毒 (IAV) 对公共卫生和禽类生产构成巨大威胁。然而，此前人们对禽类 IAV 如何招募宿主细胞机制进行复制以及宿主如何拮抗 IAV 感染知之甚少。在这里，我们展示了 H5N1 IAV PB1 N40 蛋白的病毒-宿主互作组，并揭示了其在 IAV 感染过程中涉及数十个重要宿主基因的情况。
