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分析Cullin4-E3连接体互动体及其在流感病毒感染中的重新连接

Guillaume Dugied¹, Thibaut Douche², Melanie Dos Santos³

¹Unit of Molecular Genetics of RNA Viruses, Institut Pasteur, Paris, France; Interactomics, RNA and Immunity Laboratory, Institut Pasteur, Paris, France.

Molecular & cellular proteomics : MCP

|October 9, 2024

概括

流感A病毒感染改变了库林4-RING E3泛素酶复合体与细胞蛋白相互作用的方式. 这种重新连接会影响病毒复制,并为抗病毒疗法提供新的点.

科学领域:

病毒学病毒学
分子细胞生物学分子细胞生物学
生物化学生物化学

背景情况:

库林4-RING E3泛素酶 (CRL4) 复合体对于流感A病毒 (IAV) 复制至关重要.
包括DDB1,DCAF11和DCAF12L1在内的CRL4复合体,介导病毒PB2亚单元的非降解性无化.
IAV感染对CRL4相互作用体的影响尚不清楚.

研究的目的:

为了研究IAV感染如何重塑CRL4复杂交互体.
在感染期间识别与CRL4组件相互作用的细胞蛋白.
探索这些相互作用对病毒复制和潜在治疗策略的功能后果.

主要方法:

与质谱学 (AP-MS) 结合的亲和性净化用于识别蛋白质-蛋白质相互作用.
分解纳米化酶试验以验证直接的CRL4组件合作伙伴.
通过siRNA介导的基因沉默来评估已识别的伴侣在IAV感染中的作用.

主要成果:

IAV感染显著改变了CRL4复杂的关联,减少了DDB1和DCAF11的相互作用,同时增加了DCAF12L1相关的蛋白质.
感染引起的重新连接会影响参与蛋白质折叠,无处不在,翻译,拼接和应激反应的细胞蛋白.
确定并验证了CRL4组件的直接合作伙伴对于IAV感染至关重要.

结论:

IAV感染动态重塑了CRL4 E3无素酶互动组.
这种重塑可能有助于创造有利于病毒复制的细胞环境.
已识别的CRL4相互作用蛋白代表了对宿主向抗病毒疗法的潜在标.

关键词:

库林4-RING E3 无素结合酶的使用亲和力净化与质谱学相结合.流感A型病毒感染者.蛋白质与蛋白质的相互作用分裂纳米化酶试验.

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