Obtention of Dengue Virus Membrane Proteins and Role for Virus Assembly.
Obtention of Dengue Virus Membrane Proteins and Role for Virus Assembly.
复制标题
登革热病毒膜蛋白的获得及其在病毒组装中的作用。
DOI：
10.1007/978-1-0716-1879-0_6
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Wang,Wei-Kung
中科院分区：
文献类型：
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作者：
Hsieh,Szu-Chia;Tsai,Wen-Yang;Wang,Wei-Kung
登革热病毒（DENV）的四种血清型属于黄病毒科黄病毒属，是人类虫媒病毒性疾病的主要病因。临床表现从登革热到登革出血热和登革休克综合征。尽管几十年来一直在努力制定针对DENV的干预策略，但没有获得许可的抗病毒药物，安全有效的疫苗仍然具有挑战性。与其他黄病毒类似，DENV颗粒的组装发生在内质网衍生的膜上；未成熟的病毒粒子进入管腔，然后在反式高尔基体中成熟，并通过秘书途径运输。黄病毒复制的一个独特特征是产生小而缓慢沉积的亚病毒颗粒，称为病毒样颗粒（vlp）。膜前蛋白（prM）和包膜蛋白(E)的共表达可以产生重组VLPs，其生物物理和抗原性与感染性病毒粒子相似，已被用于研究prM和E蛋白的功能、组装、血清诊断抗原和候选疫苗。在此之前，我们已经开发了几种检测方法，包括蔗糖缓冲超离心、蔗糖梯度超离心、膜浮选、亚细胞分离和糖苷酶消化试验，以利用DENV prM和E蛋白之间的相互作用、膜结合、亚细胞定位、糖基化模式以及VLPs和复制子颗粒的组装。从这些试验中获得的信息对我们进一步了解DENV的组装、复制周期、干预策略和发病机制具有重要意义。
The four serotypes of dengue virus (DENV), belonging to the genusFlavivirusin the familyFlaviviridae, are the leading cause of arboviral diseases in humans. The clinical presentations range from dengue fever to dengue hemorrhagic fever and dengue shock syndrome. Despite decades of efforts on developing intervention strategies against DENV, there is no licensed antiviral, and safe and effective vaccines remain challenging. Similar to other flaviviruses, the assembly of DENV particles occurs in the membranes derived from endoplasmic reticulum; immature virions bud into the lumen followed by maturation in the trans-Golgi and transport through the secretary pathway. A unique feature of flavivirus replication is the production of small and slowly sedimenting subviral particles, known as virus-like particles (VLPs). Co-expression of premembrane (prM) and envelope (E) proteins can generate recombinant VLPs, which are biophysically and antigenically similar to infectious virions and have been employed to study the function of prM and E proteins, assembly, serodiagnostic antigens, and vaccine candidates. Previously, we have developed several assays including sucrose cushion ultracentrifugation, sucrose gradient ultracentrifugation, membrane flotation, subcellular fractionation, and glycosidase digestion assay to exploit the interaction between DENV prM and E proteins, membrane association, subcellular localization, glycosylation pattern, and assembly of VLPs and replicon particles. The information derived from these assays have implications to further our understanding of DENV assembly, replication cycle, intervention strategies, and pathogenesis.