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与甲型流感病毒PB1-F2蛋白相互作用的细胞蛋白的鉴定

Identification of cellular proteins interacting with influenza A virus PB1-F2 protein.

作者信息

Guan Z H, Zhang M L, Hou P L, Duan M, Cui Y M, Wang X R

机构信息

Key Laboratory of Zoonosis Ministry of Education, Institute of Zoonosis, Jilin University, 130062 Changchun, P. R. China.

出版信息

Acta Virol. 2012;56(3):199-207. doi: 10.4149/av_2012_03_199.

Abstract

UNLABELLED

The influenza A virus (IAV) protein PB1-F2, which is encoded by an alternative ORF of the PB1 polymerase gene, has been implicated as an important virulence factor and apoptosis inducer. However, the molecular mechanism of PB1-F2 function remains elusive. In this study, eight cellular proteins were identified as potential PB1-F2 interacting partners using the yeast two-hybrid system. Two positive candidate proteins, guanine nucleotide binding protein (G protein) beta polypeptide 2 (Gβ2) and macrophage migration inhibitory factor (MIF), were selected to be further characterized. The interaction of MIF and Gβ2 with PB1-F2 was confirmed by both GST pull-down and co-immunoprecipitation assays. Confocal laser microscopy showed that the interaction between PB1-F2 and the two cellular proteins occurred in the cytoplasm. The novel interactions between PB1-F2 and host proteins provide further pieces of evidence in the investigation of the pathogenic mechanism of IAV.

KEYWORDS

influenza A virus; PB1-F2; yeast two-hybrid; protein-protein interaction.

摘要

未标记

甲型流感病毒(IAV)的PB1-F2蛋白由PB1聚合酶基因的一个可变开放阅读框编码,被认为是一种重要的毒力因子和凋亡诱导剂。然而,PB1-F2功能的分子机制仍不清楚。在本研究中,利用酵母双杂交系统鉴定出8种细胞蛋白作为潜在的PB1-F2相互作用伙伴。选择了两种阳性候选蛋白,鸟嘌呤核苷酸结合蛋白(G蛋白)β多肽2(Gβ2)和巨噬细胞迁移抑制因子(MIF)进行进一步表征。通过GST下拉和共免疫沉淀试验证实了MIF和Gβ2与PB1-F2的相互作用。共聚焦激光显微镜显示PB1-F2与这两种细胞蛋白之间的相互作用发生在细胞质中。PB1-F2与宿主蛋白之间的新相互作用为IAV致病机制的研究提供了更多证据。

关键词

甲型流感病毒;PB1-F2;酵母双杂交;蛋白质-蛋白质相互作用。

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