拉沙病毒
生物
聚合酶
RNA聚合酶
病毒学
相互作用体
RNA依赖性RNA聚合酶
可药性
蛋白质组
RNA聚合酶Ⅱ
核糖核酸
计算生物学
病毒
遗传学
DNA
基因表达
基因
发起人
作者
Jingru Fang,Colette Pietzsch,Haydar Witwit,George Tsaprailis,Gogce Crynen,Kelvin Frank Cho,Alice Y. Ting,Alexander Bukreyev,Erica Ollmann Saphire,Juan Carlos de la Torre
标识
DOI:10.1073/pnas.2201208119
摘要
Completion of the Lassa virus (LASV) life cycle critically depends on the activities of the virally encoded, RNA-dependent RNA polymerase in replication and transcription of the viral RNA genome in the cytoplasm of infected cells. The contribution of cellular proteins to these processes remains unclear. Here, we applied proximity proteomics to define the interactome of LASV polymerase in cells under conditions that recreate LASV RNA synthesis. We engineered a LASV polymerase-biotin ligase (TurboID) fusion protein that retained polymerase activity and successfully biotinylated the proximal proteome, which allowed the identification of 42 high-confidence LASV polymerase interactors. We subsequently performed a small interfering RNA (siRNA) screen to identify those interactors that have functional roles in authentic LASV infection. As proof of principle, we characterized eukaryotic peptide chain release factor subunit 3a (eRF3a/GSPT1), which we found to be a proviral factor that physically associates with LASV polymerase. Targeted degradation of GSPT1 by a small-molecule drug candidate, CC-90009, resulted in strong inhibition of LASV infection in cultured cells. Our work demonstrates the feasibility of using proximity proteomics to illuminate and characterize yet-to-be-defined host-pathogen interactome, which can reveal new biology and uncover novel targets for the development of antivirals against highly pathogenic RNA viruses.
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