Selective Inhibitor of Nuclear Export (SINE) Compounds Alter New World Alphavirus Capsid Localization and Reduce Viral Replication in Mammalian Cells.
Selective Inhibitor of Nuclear Export (SINE) Compounds Alter New World Alphavirus Capsid Localization and Reduce Viral Replication in Mammalian Cells.
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DOI：
10.1371/journal.pntd.0005122
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发表时间：
2016-11
影响因子：
3.8
通讯作者：
Kehn-Hall K
中科院分区：
文献类型：
--
作者：
Lundberg L;Pinkham C;de la Fuente C;Brahms A;Shafagati N;Wagstaff KM;Jans DA;Tamir S;Kehn-Hall K
新世界甲病毒委内瑞拉马脑炎病毒（VEEV）的衣壳结构蛋白与宿主核转运蛋白importin α/β1和CRM 1相互作用。新型选择性核输出抑制剂（SINE）化合物KPT-185、KPT-335（verdinexor）和KPT-350以类似于典型抑制剂Leptomycin B的方式靶向宿主的主要核输出蛋白CRM 1。Leptomycin B的一个主要限制是其与CRM 1的不可逆结合; SINE化合物缓解了这种结合，因为它们是缓慢可逆的。与非活性化合物KPT-301相比，用这些化合物化学抑制CRM 1增强了衣壳在细胞核中的定位，如免疫荧光共聚焦显微镜所示。细胞外与细胞内病毒RNA的差异以及无细胞上清液中衣壳的减少表明抑制剂影响病毒组装，这导致病毒滴度降低。病毒复制的减少是使用酶标记的病毒和通过空斑试验证实的。SINE化合物对VEEV TC83_Cm没有影响，VEEV TC83_Cm编码一种无法进入细胞核的突变形式的衣壳。在KPT-185存在下连续传代VEEV导致衣壳的核定位和核输出信号内的突变。最后，SINE化合物治疗还降低了相关的东部和西部马脑炎病毒的病毒滴度，表明CRM 1与新世界甲病毒属的衣壳蛋白保持共同的相互作用。我们的数据表明，新型选择性核输出抑制剂（SINE）化合物减少了三种相关的新世界甲病毒（VEEV、EEEV和WEEV）的病毒复制，表明CRM 1对其生命周期有帮助。新型CRM 1抑制剂具有较大的选择性指数，代表了一种潜在的泛抗病毒治疗方法，其靶向宿主的转运蛋白，这些转运蛋白被新世界甲病毒劫持。
The capsid structural protein of the New World alphavirus, Venezuelan equine encephalitis virus (VEEV), interacts with the host nuclear transport proteins importin α/β1 and CRM1. Novel selective inhibitor of nuclear export (SINE) compounds, KPT-185, KPT-335 (verdinexor), and KPT-350, target the host’s primary nuclear export protein, CRM1, in a manner similar to the archetypical inhibitor Leptomycin B. One major limitation of Leptomycin B is its irreversible binding to CRM1; which SINE compounds alleviate because they are slowly reversible. Chemically inhibiting CRM1 with these compounds enhanced capsid localization to the nucleus compared to the inactive compound KPT-301, as indicated by immunofluorescent confocal microscopy. Differences in extracellular versus intracellular viral RNA, as well as decreased capsid in cell free supernatants, indicated the inhibitors affected viral assembly, which led to a decrease in viral titers. The decrease in viral replication was confirmed using a luciferase-tagged virus and through plaque assays. SINE compounds had no effect on VEEV TC83_Cm, which encodes a mutated form of capsid that is unable to enter the nucleus. Serially passaging VEEV in the presence of KPT-185 resulted in mutations within the nuclear localization and nuclear export signals of capsid. Finally, SINE compound treatment also reduced the viral titers of the related eastern and western equine encephalitis viruses, suggesting that CRM1 maintains a common interaction with capsid proteins across the New World alphavirus genus. Our data demonstrate that novel selective inhibitor of nuclear export (SINE) compounds reduced viral replication of three related New World alphaviruses, VEEV, EEEV, and WEEV, indicating that CRM1 is instrumental to their life cycle. The novel CRM1 inhibitors have a large selective index and represent a potential pan-antiviral therapeutic that targets the host’s transport proteins, which are hijacked by the New World alphaviruses.
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影响因子：
6.2
作者：
Hayakawa, Kazuhide;Arai, Ken;Lo, Eng H.
通讯作者：
Lo, Eng H.
影响因子：
29.4
作者：
Azmi AS;Aboukameel A;Bao B;Sarkar FH;Philip PA;Kauffman M;Shacham S;Mohammad RM
通讯作者：
Mohammad RM
影响因子：
3.7
作者：
Kehn-Hall K;Narayanan A;Lundberg L;Sampey G;Pinkham C;Guendel I;Van Duyne R;Senina S;Schultz KL;Stavale E;Aman MJ;Bailey C;Kashanchi F
通讯作者：
Kashanchi F
影响因子：
5.4
作者：
Aguilar, Patricia V.;Leung, Lawrence W.;Basler, Christopher F.
通讯作者：
Basler, Christopher F.
影响因子：
5.4
作者：
Atasheva, Svetlana;Kim, Dal Young;Frolov, Ilya
通讯作者：
Frolov, Ilya