A Physical Interaction Network of Dengue Virus and Human Proteins
A Physical Interaction Network of Dengue Virus and Human Proteins
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DOI：
10.1074/mcp.m111.012187-1
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发表时间：
2011-12-01
影响因子：
7
通讯作者：
Lacount, Douglas J.
中科院分区：
文献类型：
--
作者：
Khadka, Sudip;Vangeloff, Abbey D.;Lacount, Douglas J.
登革病毒（DENV）是一种新出现的能够在人类中引起严重疾病的蚊子传播的病原体，其与宿主细胞因子相互作用以创造更有利的复制环境。然而，迄今为止，DENV与人类蛋白质之间的相互作用很少被报道。为了鉴定DENV-人蛋白质相互作用，我们使用高通量酵母双杂交测定来针对人肝脏活化结构域文库筛选10种DENV蛋白质。从45个包含全长病毒基因或部分重叠基因片段的DNA结合结构域克隆中，我们鉴定了DENV与人类蛋白质之间的139种相互作用，其中绝大多数是新的。这些相互作用涉及105种人类蛋白质，包括6种先前与DENV感染有关的蛋白质和45种与其他病毒复制有关的蛋白质。具有与补体和凝血级联、中心体和细胞骨架相关的功能的人蛋白质在DENV相互作用伴侣中富集。为了确定细胞蛋白是否是DENV感染所必需的，我们使用小干扰RNA来抑制它们的表达。靶向的12种蛋白质中的6种（CALR、DDX 3X、ERC 1、GOLGA 2、TRIP 11和UBE 2 I）导致DENV复制子的复制显着减少。我们进一步表明，钙网蛋白与病毒dsRNA和病毒NS 3和NS 5蛋白在DENV感染的细胞中共定位，这与钙网蛋白在DENV复制中的直接作用一致。与DENV相互作用的人类蛋白质的平均度和介数显著高于偶然预期，这为病毒优先靶向在人类蛋白质相互作用网络中占据中心位置的细胞蛋白质的假设提供了额外的支持。这项研究为进一步研究人类蛋白在DENV感染中的作用提供了一个有价值的起点。Molecular & Cellular Proteomics 10：10.1074/mcp.M111.012187，1-16，2011.
Dengue virus (DENV), an emerging mosquito-transmitted pathogen capable of causing severe disease in humans, interacts with host cell factors to create a more favorable environment for replication. However, few interactions between DENV and human proteins have been reported to date. To identify DENV-human protein interactions, we used high-throughput yeast two-hybrid assays to screen the 10 DENV proteins against a human liver activation domain library. From 45 DNA-binding domain clones containing either full-length viral genes or partially overlapping gene fragments, we identified 139 interactions between DENV and human proteins, the vast majority of which are novel. These interactions involved 105 human proteins, including six previously implicated in DENV infection and 45 linked to the replication of other viruses. Human proteins with functions related to the complement and coagulation cascade, the centrosome, and the cytoskeleton were enriched among the DENV interaction partners. To determine if the cellular proteins were required for DENV infection, we used small interfering RNAs to inhibit their expression. Six of 12 proteins targeted (CALR, DDX3X, ERC1, GOLGA2, TRIP11, and UBE2I) caused a significant decrease in the replication of a DENV replicon. We further showed that calreticulin colocalized with viral dsRNA and with the viral NS3 and NS5 proteins in DENV-infected cells, consistent with a direct role for calreticulin in DENV replication. Human proteins that interacted with DENV had significantly higher average degree and betweenness than expected by chance, which provides additional support for the hypothesis that viruses preferentially target cellular proteins that occupy central position in the human protein interaction network. This study provides a valuable starting point for additional investigations into the roles of human proteins in DENV infection. Molecular & Cellular Proteomics 10: 10.1074/mcp.M111.012187, 1-16, 2011.