Identification and characterization of prohibitin as a receptor protein mediating DENV-2 entry into insect cells
Identification and characterization of prohibitin as a receptor protein mediating DENV-2 entry into insect cells
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DOI：
10.1016/j.virol.2010.07.015
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发表时间：
2010-10-10
期刊：
影响因子：
3.7
通讯作者：
Smith, Duncan R.
中科院分区：
文献类型：
--
作者：
Kuadkitkan, Atichat;Wikan, Nitwara;Smith, Duncan R.
登革热主要由伊蚊属的蚊子传播。尽管进行了大量的研究，但没有一种昆虫登革热病毒受体蛋白被明确地鉴定和表征。利用多种分离方法和病毒覆盖蛋白结合试验，我们在两种蚊子和两种蚊子细胞系中分离出一种35 kDa的对登革热血清型2 （DENV-2）感染敏感的蛋白。质谱鉴定该蛋白为禁止蛋白，一种在真核细胞中广泛表达的强保守蛋白。抗体介导的感染抑制和siRNA介导的抑制prohibition表达显著降低了埃及伊蚊和白纹伊蚊细胞系的感染水平和随后的病毒产量。共聚焦显微镜显示，禁止蛋白与denv - 2e蛋白在细胞内共定位，共免疫沉淀证实了禁止蛋白与登革热E相互作用。禁止蛋白是第一个在昆虫细胞中表达的登革病毒受体蛋白。(C) 2010爱思唯尔公司版权所有。
Dengue is transmitted primarily by mosquitoes of the Aedes genus. Despite a number of studies, no insect dengue virus receptor protein has been clearly identified and characterized. Using a number of separation methodologies and virus overlay protein binding assays we identified a 35 kDa protein that segregated with susceptibility to dengue serotype 2 (DENV-2) infection in two mosquito species and two mosquito cell lines. Mass spectroscopy identified the protein to be prohibitin, a strongly conserved and ubiquitously expressed protein in eukaryotic cells. Antibody mediated inhibition of infection and siRNA mediated knockdown of prohibitin expression significantly reduced infection levels and subsequent virus production in both Aedes aegypti and Aedes albopictus cell lines. Confocal microscopy showed a significant degree of intracellular colocalization between prohibitin and DENV-2 E protein, and coimmunoprecipitation confirmed that prohibitin interacts with dengue E. Prohibitin is the first characterized insect cell expressed dengue virus receptor protein. (C) 2010 Elsevier Inc. All rights reserved.