两株小鼠致病性差异显著的H5N1禽流感病毒PA蛋白差异互作宿主蛋白研究

Study on the Different Interactions between PA Protein of Two H5N1 Avian Influenza Viruses with Markedly Different Pathogenicity in Mice and Host Proteins

为了探索引起H5N1高致病性禽流感病毒(highly pathogenic avian influenza virus,HPAIV)致病性差异的宿主因子,选取了一对主要由PA基因造成在小鼠上致病性差异的H5N1禽流感病毒(CK10与GS10),通过病毒感染A549细胞,利用PA抗体进行免疫沉淀(immunoprecipitation,IP)试验以筛选与PA互作的宿主蛋白,并进行LC-MS/MS鉴定。结果发现与低致病性的GS10相比,共有160种宿主蛋白能够特异性与高致病性CK10病毒PA蛋白结合。对这160种蛋白进行生物信息学分析,通过Gene Ontology(GO)注释分析发现这些蛋白主要参与翻译、基因表达、病毒转录和病毒感染等生物学进程,通过KEGG通路分析发现这些蛋白主要参与的细胞通路包括翻译、传染病与信号转导等。此后,利用免疫共沉淀技术(co-immunoprecipitation,Co-IP),证实所筛选的宿主蛋白eEF1A1能够特异性地与CK10病毒PA蛋白相互作用。本研究筛选到了致病性不同的禽流感病毒的差异互作宿主蛋白,可为进一步了解流感病毒复杂的致病机制提供参考。

To explore the host factors that contribute to the pathogenicity of the H5N1 influenza virus,Two H5N1 avian influenza viruses with different pathogenicity mainly determined by PA(CK10 and GS10)were used in this study.A549 cells were infected with the two viruses and immunoprecipitation(IP)experiments were performed by using PA antibodies subsequently to screen host proteins that interact with PA.The host proteins were further identified by LC-MS/MS.As a result,compared with the low pathogenic GS10,there were 160 host proteins interacting with the high pathogenic CK10 specifically.Bioinformatics analysis was employed targeting on these 160 proteins subsequently.Gene Ontology(GO)annotation analysis showed that these proteins mainly involved in the biological processes of translation,gene expression,viral transcription and viral infection.KEGG pathway analysis revealed that these proteins mainly participate in the cell pathways of translation,infectious diseases and signal transduction.Using co-immunoprecipitation(Co-IP),host protein eEF1A1 was identified to interact with CK10 PA protein specifically.Therefore,this study successfully screened out the differential interactome data of PA protein from different pathogenicity H5N1 influenza viruses.It may play pivotal roles in elucidating the complicated pathogenesis of H5N1 influenza virus.