Barrera Michael D, Callahan Victoria, Akhrymuk Ivan, Bhalla Nishank, Zhou Weidong, Campbell Catherine, Narayanan Aarthi, Kehn-Hall Kylene
National Center for Biodefense and Infectious Diseases, School of Systems Biology, George Mason University, Manassas, VA 20110, USA.
Department of Biomedical Sciences and Pathobiology, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Pathogens. 2021 Mar 2;10(3):283. doi: 10.3390/pathogens10030283.
Alphaviruses are a genus of the family and are widely distributed across the globe. Venezuelan equine encephalitis virus (VEEV) and eastern equine encephalitis virus (EEEV), cause encephalitis and neurological sequelae while chikungunya virus (CHIKV) and Sindbis virus (SINV) cause arthralgia. There are currently no approved therapeutics or vaccines available for alphaviruses. In order to identify novel therapeutics, a V5 epitope tag was inserted into the N-terminus of the VEEV E2 glycoprotein and used to identify host-viral protein interactions. Host proteins involved in protein folding, metabolism/ATP production, translation, cytoskeleton, complement, vesicle transport and ubiquitination were identified as VEEV E2 interactors. Multiple inhibitors targeting these host proteins were tested to determine their effect on VEEV replication. The compound HA15, a GRP78 inhibitor, was found to be an effective inhibitor of VEEV, EEEV, CHIKV, and SINV. VEEV E2 interaction with GRP78 was confirmed through coimmunoprecipitation and colocalization experiments. Mechanism of action studies found that HA15 does not affect viral RNA replication but instead affects late stages of the viral life cycle, which is consistent with GRP78 promoting viral assembly or viral protein trafficking.
甲病毒是该科的一个属，广泛分布于全球。委内瑞拉马脑炎病毒（VEEV）和东部马脑炎病毒（EEEV）可引起脑炎和神经后遗症，而基孔肯雅病毒（CHIKV）和辛德毕斯病毒（SINV）可引起关节痛。目前尚无针对甲病毒的获批治疗方法或疫苗。为了确定新型治疗方法，将一个V5表位标签插入VEEV E2糖蛋白的N端，用于鉴定宿主-病毒蛋白相互作用。参与蛋白质折叠、代谢/ATP生成、翻译、细胞骨架、补体、囊泡运输和泛素化的宿主蛋白被鉴定为VEEV E2相互作用蛋白。测试了多种针对这些宿主蛋白的抑制剂，以确定它们对VEEV复制的影响。化合物HA15是一种GRP78抑制剂，被发现是VEEV、EEEV、CHIKV和SINV的有效抑制剂。通过免疫共沉淀和共定位实验证实了VEEV E2与GRP78的相互作用。作用机制研究发现，HA15不影响病毒RNA复制，而是影响病毒生命周期的后期阶段，这与GRP78促进病毒组装或病毒蛋白运输一致。
