Unique Requirement for ESCRT Factors in Flavivirus Particle Formation on the Endoplasmic Reticulum
Unique Requirement for ESCRT Factors in Flavivirus Particle Formation on the Endoplasmic Reticulum
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DOI：
10.1016/j.celrep.2016.07.068
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发表时间：
2016-08-30
期刊：
影响因子：
8.8
通讯作者：
Morita, Eiji
中科院分区：
文献类型：
--
作者：
Tabata, Keisuke;Arimoto, Masaru;Morita, Eiji
黄病毒感染诱导内质网（ER）膜重排，产生一个室，用于病毒基因组的复制和病毒颗粒的组装。使用定量质谱法，我们确定了几种ESCRT（运输所需的内体分选复合体）蛋白，它们被招募到内质网上的病毒复制位点。系统小干扰RNA （siRNA）筛选显示，单次缺失TSG101或共同缺失ESCRT-III蛋白的特定组合可显著降低登革热病毒和日本脑炎病毒的释放，导致滴度降低>= 1000倍。相比之下，释放不受一些核心escrt消耗的影响，包括VPS4。将ESCRT蛋白重新引入sirna缺失的细胞，揭示了ESCRT蛋白之间的相互作用对黄病毒出芽至关重要。电镜研究表明，CHMP2和CHMP4蛋白在内质网处的膜变形中起直接作用。因此，在黄病毒感染期间，ESCRT的一个独特的特异性亚群有助于内质网膜的生物发生。
Flavivirus infection induces endoplasmic reticulum (ER) membrane rearrangements to generate a compartment for replication of the viral genome and assembly of viral particles. Using quantitative mass spectrometry, we identified several ESCRT (endosomal sorting complex required for transport) proteins that are recruited to sites of virus replication on the ER. Systematic small interfering RNA (siRNA) screening revealed that release of both dengue virus and Japanese encephalitis virus was dramatically decreased by single depletion of TSG101 or co-depletion of specific combinations of ESCRT-III proteins, resulting in >= 1,000-fold titer reductions. By contrast, release was unaffected by depletion of some core ESCRTs, including VPS4. Reintroduction of ESCRT proteins to siRNA-depleted cells revealed interactions among ESCRT proteins that are crucial for flavivirus budding. Electron-microscopy studies revealed that the CHMP2 and CHMP4 proteins function directly in membrane deformation at the ER. Thus, a unique and specific subset of ESCRT contributes to ER membrane biogenesis during flavivirus infection.