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标题：Ebola and Marburg virus VP35 coiled-coil validated as antiviral target by tripartite split-GFP complementation
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作者：Luca Zinzula ; Angela Maria Mereu ; Massimiliano Orsini 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：11
页码：1-24
DOI：10.1016/j.isci.2022.105354
语种：English
出版社：Elsevier
摘要：SummaryEbola virus (EBOV) and Marburg virus (MARV) are highly pathogenic viruses in humans, against which approved antivirals are lacking. During EBOV and MARV infection, coiled-coil mediated oligomerization is essential for the virion protein 35 (VP35) polymerase co-factor function and type I interferon antagonism, making VP35 coiled-coil an elective drug target. We established a tripartite split-green fluorescent protein (GFP) fluorescence complementation (FC) system based on recombinant GFP-tagged EBOV and MARV VP35, which probes VP35 coiled-coil assembly by monitoring fluorescence onE. colicolonies, orin vitroin 96/384-multiwell. Oligomerization-defective VP35 mutants showed that correct coiled-coil knobs-into-holes pairing within VP35 oligomer is pre-requisite for GFP tags and GFP detector to reconstitute fluorescing full-length GFP. The method was validated by screening a small compound library, which identified Myricetin and 4,5,6,7-Tetrabromobenzotriazole as inhibitors of EBOV and MARV VP35 oligomerization-dependent FC with low-micromolar IC50values. These findings substantiate the VP35 coiled-coil value as antiviral target.Graphical abstractDisplay OmittedHighlights•Ebola and Marburg virus VP35 oligomerize via trimeric and tetrameric coiled-coil•VP35 coiled-coil assembly triggers fluorescence of a tripartite split-GFP system•Mutations perturbing VP35 coiled-coil hamper split-GFP complementation•Myricetin and TBBT inhibit split-GFP complementation mediated by VP35 coiled-coilDrugs; Virology.