New insights into the role of endosomal proteins for African swine fever virus infection.
New insights into the role of endosomal proteins for African swine fever virus infection.
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DOI：
10.1371/journal.ppat.1009784
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发表时间：
2022-01
期刊：
影响因子：
6.7
通讯作者：
Alonso C
中科院分区：
文献类型：
--
作者：
Cuesta-Geijo MÁ;García-Dorival I;Del Puerto A;Urquiza J;Galindo I;Barrado-Gil L;Lasala F;Cayuela A;Sorzano COS;Gil C;Delgado R;Alonso C
非洲猪瘟病毒（African Swine Fever Virus，ASFV）的感染周期始于病毒吸附并进入宿主细胞。然后，病毒通过网格蛋白/发动蛋白介导的内吞作用和巨胞饮作用内化。与其他病毒类似，ASF病毒粒子随后被内化并并入内吞途径。虽然内体成熟需要腔酸化，但pH的降低作用于病毒粒子的多层结构，溶解外部衣壳。在去衣壳后，病毒内膜暴露以与晚期内体的界膜相互作用以进行融合。然后，病毒融合对于进入的病毒体从内体进入细胞质是必要的，然而，这仍然是感染的有趣且必需的过程，特别是对于病毒核酸进入细胞质进行复制。ASFV蛋白E248 R和E199 L位于暴露的病毒内膜，可能涉及融合步骤。显示了这些病毒蛋白与细胞内体蛋白如尼曼-匹克C 1型（NPC 1）和溶酶体膜蛋白（Lamp-1和Lamp-2）之间的相互作用。此外，这些蛋白质的沉默削弱了ASFV感染。还观察到，使用CRISPR敲除NPC 1的细胞危害ASFV感染，并且病毒核心的进展和内体退出在病毒进入时在内体内被阻止。这些结果表明，ASFV蛋白与一些内体蛋白的相互作用可能是重要的膜融合步骤。除此之外，NPC 1 KO细胞中ASFV感染性和复制的降低伴随着更少和更小的病毒工厂。我们的研究结果铺平了道路，了解蛋白质的内体膜在ASFV感染的作用。非洲猪瘟病毒（ASFV）引起猪和野猪的致命疾病，该疾病曾在非洲流行，但近年来已蔓延到欧洲，亚洲和大洋洲，造成严重的社会经济影响。ASFV通过内吞作用进入细胞，并适应内体条件以获得感染性。病毒内膜与内体膜的融合是病毒DNA进入细胞质以开始复制所必需的。我们已经发现ASF病毒粒子内膜蛋白E248 R和E199 L与内体蛋白Niemann Pick C1（NPC 1）和溶酶体膜蛋白（Lamp）-1和-2相互作用。并且，似乎是ASF病毒粒子的内体运输所必需的，内体运输和在细胞进入过程中退出到细胞质。这些分子调节胆固醇从内体流向内质网，并且似乎对病毒感染周期很重要。在沉默和敲除细胞中，ASFV感染在早期和晚期受到影响。在空细胞中，病毒粒子进入和进入时通过内体途径的进展被阻止，并且几个病毒核心保留在晚期内体，而没有进入融合期以用于胞质退出。这些结果提供了新的见解内体蛋白的作用ASFV感染。
African swine fever virus (ASFV) infectious cycle starts with the viral adsorption and entry into the host cell. Then, the virus is internalized via clathrin/dynamin mediated endocytosis and macropinocytosis. Similar to other viruses, ASF virion is then internalized and incorporated into the endocytic pathway. While the endosomal maturation entails luminal acidification, the decrease in pH acts on the multilayer structure of the virion dissolving the outer capsid. Upon decapsidation, the inner viral membrane is exposed to interact with the limiting membrane of the late endosome for fusion. Viral fusion is then necessary for the egress of incoming virions from endosomes into the cytoplasm, however this remains an intriguing and yet essential process for infection, specifically for the egress of viral nucleic acid into the cytoplasm for replication. ASFV proteins E248R and E199L, located at the exposed inner viral membrane, might be implicated in the fusion step. An interaction between these viral proteins and cellular endosomal proteins such as the Niemann-Pick C type 1 (NPC1) and lysosomal membrane proteins (Lamp-1 and -2) was shown. Furthermore, the silencing of these proteins impaired ASFV infection. It was also observed that NPC1 knock-out cells using CRISPR jeopardized ASFV infection and that the progression and endosomal exit of viral cores was arrested within endosomes at viral entry. These results suggest that the interactions of ASFV proteins with some endosomal proteins might be important for the membrane fusion step. In addition to this, reductions on ASFV infectivity and replication in NPC1 KO cells were accompanied by fewer and smaller viral factories. Our findings pave the way to understanding the role of proteins of the endosomal membrane in ASFV infection. African swine fever virus (ASFV) causes a deadly disease of pigs and wild boars that was endemic in Africa but has spread in recent years to Europe, Asia and Oceania with a high socioeconomic impact. ASFV enters the cell by endocytosis and has adapted to the endosomal conditions to acquire infectivity. Fusion of the internal viral membrane with the endosomal membrane is required for the exit of viral DNA into the cytoplasm to start replication. We have found that ASF virion internal membrane proteins E248R and E199L interact with the endosomal proteins Niemann Pick C1 (NPC1) and lysosomal membrane proteins (Lamp)-1 and -2. And, appear to be required for endosomal trafficking of ASF virions endosomal traffic and exit to the cytoplasm in the cell entry process. These molecules act regulating cholesterol flux from the endosome to the endoplasmic reticulum, and appear to be important for the viral infection cycle. In silenced and knockout cells, ASFV infection was affected at early and later stages. In null cells, virion entry and progression through the endosomal pathway at entry was arrested and several viral cores were retained at late endosomes without entering the fusion phase for cytoplasmic exit. These results provide new insights into the role of endosomal proteins for ASFV infection.
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影响因子：
64.8
作者：
Cote, Marceline;Misasi, John;Ren, Tao;Bruchez, Anna;Lee, Kyungae;Filone, Claire Marie;Hensley, Lisa;Li, Qi;Ory, Daniel;Chandran, Kartik;Cunningham, James
通讯作者：
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影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Hiscox, Julian A.
影响因子：
4.3
作者：
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