Rapid Degradation Pathways of Host Proteins During HCMV Infection Revealed by Quantitative Proteomics.
Rapid Degradation Pathways of Host Proteins During HCMV Infection Revealed by Quantitative Proteomics.
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DOI：
10.3389/fcimb.2020.578259
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发表时间：
2020
影响因子：
5.7
通讯作者：
Weekes MP
中科院分区：
文献类型：
--
作者：
Lin KM;Nightingale K;Soday L;Antrobus R;Weekes MP
人巨细胞病毒（HCMV）是免疫功能低下个体和新生儿的重要病原体，也是病毒免疫逃逸的范例。我们以前开发了一种定量蛋白质组学方法，鉴定了133种在HCMV感染早期降解的蛋白质，包括已知和新型的抗病毒因子。大多数被MG 132从降解中拯救出来，已知MG 132除了抑制蛋白酶体之外还抑制溶酶体组织蛋白酶。宿主蛋白质降解的精确机制的全球定义对于提高我们对病毒生物学的理解和告知新的抗病毒治疗策略都很重要。因此，我们开发和优化了一个多路比较蛋白质组学分析，使用选择性蛋白酶体抑制剂硼替佐米除了MG 132，提供一个全球性的蛋白质降解机制的观点。在被MG 132从降解中拯救的蛋白质中，34-47个蛋白质也被硼替佐米拯救，这表明HCMV采用的蛋白质降解的主要机制是通过蛋白酶体，并且降解的替代途径仍然很重要。我们的方法和数据将有助于提高对HCMV和其他病毒的机制理解，并为进一步分析提供候选限制因子的短名单。
Human cytomegalovirus (HCMV) is an important pathogen in immunocompromised individuals and neonates, and a paradigm for viral immune evasion. We previously developed a quantitative proteomic approach that identified 133 proteins degraded during the early phase of HCMV infection, including known and novel antiviral factors. The majority were rescued from degradation by MG132, which is known to inhibit lysosomal cathepsins in addition to the proteasome. Global definition of the precise mechanisms of host protein degradation is important both to improve our understanding of viral biology, and to inform novel antiviral therapeutic strategies. We therefore developed and optimized a multiplexed comparative proteomic analysis using the selective proteasome inhibitor bortezomib in addition to MG132, to provide a global mechanistic view of protein degradation. Of proteins rescued from degradation by MG132, 34–47 proteins were also rescued by bortezomib, suggesting both that the predominant mechanism of protein degradation employed by HCMV is via the proteasome, and that alternative pathways for degradation are nevertheless important. Our approach and data will enable improved mechanistic understanding of HCMV and other viruses, and provide a shortlist of candidate restriction factors for further analysis.