与PRRSV nsp11互作的宿主细胞蛋白鉴定及生物信息学分析

发布时间：2018-07-03 15:56

本文选题：猪繁殖与呼吸综合征病毒 + 非结构蛋白　；参考：《微生物学通报》2017年12期

【摘要】：【目的】研究猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)nsp11与宿主细胞蛋白之间的相互作用,对于揭示nsp11在病毒复制过程中发挥的功能至关重要。【方法】在病毒感染细胞的基础上,利用nsp11的单克隆抗体,采用免疫沉淀结合串联质谱的方法,筛选与PRRSV nsp11相互作用的宿主细胞蛋白,并对所筛选出的宿主细胞蛋白进行了GO注释、COG注释和KEGG代谢通路注释;选取筛选出的宿主细胞蛋白IRAK1,利用免疫共沉淀技术和激光共聚焦技术鉴定其与nsp11之间的相互作用。【结果】与空白对照组相比,病毒感染组中出现3条差异带;经质谱分析共筛选得到了201个与nsp11相互作用的宿主细胞蛋白,分别与蛋白质代谢、细胞信号通路转导以及病原致病性等密切相关;在生物信息学分析的基础上,实验验证了nsp11确与宿主细胞蛋白IRAK1进行相互作用。【结论】鉴定出与PRRSV nsp11相互作用的宿主细胞蛋白,生物信息学分析显示它们在病毒的复制和致病过程中发挥重要作用。研究结果为探究nsp11的生物学功能指明了方向,也为研究宿主细胞蛋白与病毒蛋白间的相互作用及其调控病毒复制和致病性的分子机制奠定了基础。
[Abstract]:[objective] to study the interaction between porcine reproductive and respiratory syndrome virus (PRRSv) nsp11 and host cell proteins, in order to reveal the function of nsp11 in viral replication. The host cell proteins interacting with PRRSv nsp11 were screened by immunoprecipitation and tandem mass spectrometry using monoclonal antibody of nsp11. The selected host cell proteins were annotated by go and KEGG metabolic pathway. The host cell protein IRAK1 was selected and the interaction between IRAK1 and nsp11 was identified by immunoprecipitation and laser confocal technique. [results] compared with the control group, there were three different bands in the viral infection group. A total of 201 host cell proteins interacting with nsp11 were screened by mass spectrometry, which were closely related to protein metabolism, cell signal pathway transduction and pathogenicity. It was verified that nsp11 interacted with host cell protein IRAK1. [conclusion] the host cell proteins interacting with PRRSv nsp11 were identified. Bioinformatics analysis showed that they played an important role in the replication and pathogenesis of the virus. The results provide a basis for the study of the biological function of nsp11 and the interaction between host cell protein and viral protein and the molecular mechanism of regulating viral replication and pathogenicity.
【作者单位】：中国农业大学动物医学院农业部动物流行病学重点实验室;
【基金】：国家科技攻关计划项目(No.2015BAD12B01-2)~~
【分类号】：S852.651

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