Ebolavirus is internalized into host cells via macropinocytosis in a viral glycoprotein-dependent manner.
Ebolavirus is internalized into host cells via macropinocytosis in a viral glycoprotein-dependent manner.
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DOI：
10.1371/journal.ppat.1001121
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发表时间：
2010-09-23
期刊：
影响因子：
6.7
通讯作者：
Kawaoka Y
中科院分区：
文献类型：
--
作者：
Nanbo A;Imai M;Watanabe S;Noda T;Takahashi K;Neumann G;Halfmann P;Kawaoka Y
埃博拉病毒（EBOV）是一种有包膜的单链负义RNA病毒，可引起严重的出血热，在人类和非人灵长类动物中的死亡率高达90%。先前的研究表明网格蛋白或小窝介导的内吞作用在EBOV进入中的作用;然而，埃博拉病毒病毒粒子是长的丝状颗粒，其大于质膜内陷，其表征网格蛋白或小窝介导的内吞作用。因此，对EBOV进入的机制仍然知之甚少。为了更好地了解埃博拉病毒的进入，我们用荧光标记的、生物学上含有的埃博拉病毒和埃博拉病毒样颗粒（埃博拉病毒VLP）进行了内化研究，这两种病毒在形态上都类似于真实的埃博拉病毒。我们通过实时分析这些荧光标记的埃博拉病毒颗粒来研究埃博拉病毒内化的机制，发现它们的内化不依赖于网格蛋白或小窝介导的内吞作用，但它们与分选连接蛋白（SNX）5（巨胞饮特异性内体（巨胞饮体）的标志物）共定位。此外，埃博拉病毒病毒粒子的内化加速了巨胞饮特异性货物的摄取，与质膜皱褶相关，并依赖于参与巨胞饮的细胞GTP酶和激酶。具有埃博拉病毒糖蛋白（GP）的假型水疱性口炎病毒也与SNX 5共定位，其内化和感染性受到巨胞饮抑制剂的影响。综上所述，我们的数据表明，埃博拉病毒通过以GP依赖性方式刺激巨胞饮而内化到细胞中。这些发现为埃博拉病毒的生命周期提供了新的见解，并可能有助于开发埃博拉病毒感染的治疗方法。埃博拉病毒（EBOV）是一种有包膜的单链负义RNA病毒，可引起人类和非人灵长类动物的严重出血热，死亡率高。先前的研究表明网格蛋白或小窝介导的内吞作用在EBOV进入中的作用;然而，关于EBOV进入的机制仍然存在问题。在这里，我们证明了EBOV颗粒的内化是独立的网格蛋白或小窝介导的内吞作用。具体而言，我们表明内化的EBOV颗粒与巨胞饮特异性内体（巨胞饮体）共定位，并且它们的进入受到巨胞饮抑制剂治疗的负面影响。此外，埃博拉病毒粒子的内化加速了巨胞饮特异性货物的摄取，与质膜皱褶相关，并且依赖于参与巨胞饮的细胞GTP酶和激酶。我们进一步证明，具有EBOV糖蛋白（GP）的假型水泡性口炎病毒也与巨胞饮体共定位，其内化也受到巨胞饮抑制剂的影响。我们的研究结果表明，EBOV摄取到细胞中涉及巨胞饮途径，是GP依赖性的。这些发现为EBOV的生命周期提供了新的见解，并可能有助于开发EBOV感染的治疗方法。
Ebolavirus (EBOV) is an enveloped, single-stranded, negative-sense RNA virus that causes severe hemorrhagic fever with mortality rates of up to 90% in humans and nonhuman primates. Previous studies suggest roles for clathrin- or caveolae-mediated endocytosis in EBOV entry; however, ebolavirus virions are long, filamentous particles that are larger than the plasma membrane invaginations that characterize clathrin- or caveolae-mediated endocytosis. The mechanism of EBOV entry remains, therefore, poorly understood. To better understand Ebolavirus entry, we carried out internalization studies with fluorescently labeled, biologically contained Ebolavirus and Ebolavirus-like particles (Ebola VLPs), both of which resemble authentic Ebolavirus in their morphology. We examined the mechanism of Ebolavirus internalization by real-time analysis of these fluorescently labeled Ebolavirus particles and found that their internalization was independent of clathrin- or caveolae-mediated endocytosis, but that they co-localized with sorting nexin (SNX) 5, a marker of macropinocytosis-specific endosomes (macropinosomes). Moreover, the internalization of Ebolavirus virions accelerated the uptake of a macropinocytosis-specific cargo, was associated with plasma membrane ruffling, and was dependent on cellular GTPases and kinases involved in macropinocytosis. A pseudotyped vesicular stomatitis virus possessing the Ebolavirus glycoprotein (GP) also co-localized with SNX5 and its internalization and infectivity were affected by macropinocytosis inhibitors. Taken together, our data suggest that Ebolavirus is internalized into cells by stimulating macropinocytosis in a GP-dependent manner. These findings provide new insights into the lifecycle of Ebolavirus and may aid in the development of therapeutics for Ebolavirus infection. Ebolavirus (EBOV) is an enveloped, single-stranded, negative-sense RNA virus that causes severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. Previous studies suggest roles for clathrin- or caveolae-mediated endocytosis in EBOV entry; however, questions remain regarding the mechanism of EBOV entry. Here, we demonstrate that internalization of EBOV particles is independent of clathrin- or caveolae-mediated endocytosis. Specifically, we show that internalized EBOV particles co-localize with macropinocytosis-specific endosomes (macropinosomes) and that their entry is negatively affected by treatment with macropinocytosis inhibitors. Moreover, the internalization of Ebola virions accelerated the uptake of a macropinocytosis-specific cargo, was associated with plasma membrane ruffling, and was dependent on cellular GTPases and kinases involved in macropinocytosis. We further demonstrate that a pseudotyped vesicular stomatitis virus possessing the EBOV glycoprotein (GP) also co-localizes with macropinosomes and its internalization is similarly affected by macropinocytosis inhibitors. Our results indicate that EBOV uptake into cells involves the macropinocytic pathway and is GP-dependent. These findings provide new insights into the lifecycle of EBOV and may aid in the development of therapeutics for EBOV infection.
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影响因子：
3.8
作者：
Ji, X;Olinger, GG;Spear, GT
通讯作者：
Spear, GT
影响因子：
6.7
作者：
Ghigo, Eric;Kartenbeck, Juergen;Lien, Pham;Pelkmans, Lucas;Capo, Christian;Mege, Jean-Louis;Raoult, Didier
通讯作者：
Raoult, Didier
DOI：
10.1083/jcb.131.6.1435
发表时间：
1995-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Feng Y;Press B;Wandinger-Ness A
通讯作者：
Wandinger-Ness A
影响因子：
3.3
作者：
Amyere, M;Payrastre, B;Courtoy, PJ
通讯作者：
Courtoy, PJ
DOI：
10.1083/jcb.83.1.82
发表时间：
1979-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Haigler HT;McKanna JA;Cohen S
通讯作者：
Cohen S