Siriraj Center of Research Excellence for Molecular Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand; Molecular Medicine Program, Multidisciplinary Unit, Faculty of Science, Faculty of Medicine Ramathibodi Hospital, Faculty of Dentistry, Faculty of Tropical Medicine, Faculty of Graduate Studies, Mahidol University, Bangkok, Thailand.
Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, Thailand.
Virus Res. 2018 May 2;250:13-20. doi: 10.1016/j.virusres.2018.03.021. Epub 2018 Mar 30.
Dengue hemorrhagic fever is a life-threatening disease caused by the dengue virus (DENV). After DENV enters into host cells, it replicates to generate viral particles to infect other cells. DENV exploits components of the cellular trafficking pathway to achieve effective virion production. Understanding of the proteins required for this trafficking process is essential for revealing the pathogenesis of DENV infection. Coat protein complex and soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), two host protein families in the cellular trafficking pathway, were investigated to elucidate their respective roles during DENV infection. Coat proteins (COPI and COPII) and SNAREs (STX 5 and NSF) were knocked down in a DENV-infected Huh7 cells by RNA interference. Depletion of COPI and COPII, but not of STX5 and NSF, decreased DENV production in DENV-infected Huh7 cells. DENV proteins, including DENV C, prM, E, and NS1, were significantly reduced in COPI-silenced DENV-infected Huh7 cells, when compared to those of control cells. COPI also facilitated DENV production in an endothelial cell line and in all DENV serotypes, indicating the importance of coat protein complex in facilitating DENV infection.
登革出血热是由登革病毒（DENV）引起的危及生命的疾病。DENV 进入宿主细胞后，会复制产生病毒颗粒以感染其他细胞。DENV 利用细胞运输途径的成分来实现有效的病毒颗粒产生。了解该运输过程所需的蛋白质对于揭示 DENV 感染的发病机制至关重要。本研究调查了衣壳蛋白复合物和可溶性 N-乙基马来酰亚胺敏感因子附着蛋白受体（SNAREs）这两种细胞运输途径中的宿主蛋白家族，以阐明它们在 DENV 感染过程中的各自作用。通过 RNA 干扰在 DENV 感染的 Huh7 细胞中敲低衣壳蛋白（COPI 和 COPII）和 SNAREs（STX5 和 NSF）。与对照细胞相比，COPI 和 COPII 的耗竭而非 STX5 和 NSF 的耗竭降低了 DENV 感染的 Huh7 细胞中的 DENV 产量。与对照细胞相比，沉默 COPI 的 DENV 感染的 Huh7 细胞中的 DENV 蛋白（包括 DENV C、prM、E 和 NS1）明显减少。COPI 还促进了内皮细胞系和所有 DENV 血清型的 DENV 产生，表明衣壳蛋白复合物在促进 DENV 感染方面的重要性。
