Protein disulfide isomerases (PDIs) negatively regulate ebolavirus structural glycoprotein expression in the endoplasmic reticulum (ER) via the autophagy-lysosomal pathway
Protein disulfide isomerases (PDIs) negatively regulate ebolavirus structural glycoprotein expression in the endoplasmic reticulum (ER) via the autophagy-lysosomal pathway
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蛋白质二硫键异构酶（PDI）通过自噬-溶酶体途径负向调节内质网（ER）中埃博拉病毒结构糖蛋白的表达
DOI：
10.1080/15548627.2022.2031381
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发表时间：
2022-02-09
期刊：
影响因子：
13.3
通讯作者：
Zheng, Yong-Hui
中科院分区：
文献类型：
--
作者：
Wang, Bin;Zhang, Jing;Zheng, Yong-Hui
扎伊尔埃博拉病毒（EBOV）在人类和非人类灵长类动物中引起严重的出血热，具有高发病率和死亡率。EBOV感染依赖于其结构糖蛋白（GP），但GP的高水平表达也会引发细胞变圆、脱离和许多表面分子的下调，这被认为是导致其高致病性的原因。因此，EBOV进化出了一种RNA编辑机制，以减少其GP表达并增加其适应度。我们现在报道GP的表达也被蛋白二硫异构酶（pdi）在细胞的蛋白水平上抑制。虽然pdi通过在内质网（ER）中催化正确的二硫键形成来促进氧化蛋白折叠，但PDIA3/ERp57通过靶向GP半胱氨酸残基并激活未折叠蛋白反应（UPR）而对GP错误折叠产生不利影响。异常折叠的GP是er相关蛋白降解（ERAD）机制的靶标，出乎意料的是，它通过巨噬/自噬-溶酶体途径而不是蛋白酶体途径被降解。PDIA3还降低了其他埃博拉病毒的GP表达，但增加了马尔堡病毒（MARV）的GP表达，这与MARV-GP不引起细胞圆角和脱离的观察结果一致，MARV在感染过程中不通过RNA编辑来调节其GP表达。此外，其他5种pdi也具有与EBOV-GP相似的抑制活性。因此，pdi负性调节埃博拉病毒糖蛋白表达，从而通过最大化其感染而最小化其细胞效应来平衡病毒生命周期。我们认为埃博拉病毒在感染期间劫持宿主蛋白折叠和ERAD机制，通过网状吞噬来增加其适应性。缩写：3-MA: 3-甲基腺嘌呤；4-PBA: 4-phenylbutyrate;ACTB:β肌动蛋白;ATF：激活转录因子；ATG: autophagy-related;BafA1：巴霉素A1；本迪布焦埃博拉病毒；CALR: calreticulin;CANX: calnexin;CHX:环己酰亚胺;CMA：伴侣介导的自噬；ConA：康纳霉素A；CRISPR：有规则间隔的短回文重复序列簇；Cas9: crispr相关蛋白9；dsRNA：双链RNA；扎伊尔埃博拉病毒；EDEM：内质网降解增强-甘露糖苷酶样蛋白；EIF2AK3/PERK：真核翻译起始因子2 α激酶3；Env：包膜糖蛋白；ER：内质网；ERAD: er相关蛋白降解；ERN1/IRE1：内质网至核信号1；医生:糖蛋白;HA:血凝素;HDAC6：组蛋白去乙酰化酶6；嗯:high-molecular-mass;HIV-1：人类免疫缺陷病毒1型；HSPA5/BiP：热休克蛋白家族A （Hsp70）成员5；IAV：甲型流感病毒；知识产权:免疫沉淀反应;麻醉品:kifenesine;Lac: lactacystin;LAMP：溶酶体相关膜蛋白；MAN1B1/ERManI：甘露糖苷酶α 1B类成员1；MAP1LC3/LC3：微管相关蛋白1轻链3；马尔堡病毒；MLD：粘蛋白样结构域；NHK/SERPINA1: alpha1-抗胰蛋白酶变异null（香港）；NTZ: nitazoxanide;PDI：蛋白质二硫异构酶；RAVV：乌鸦病毒；雷斯顿埃博拉病毒；SARS-CoV：严重急性呼吸综合征冠状病毒；苏丹埃博拉病毒；sGP：可溶性GP；SQSTM1/p62: sequestosome 1；ssGP：可溶性小GP；TAFV: Taï森林埃博拉病毒；TIZ: tizoxanide;TGN: thapsigargin;TLD: TXN (thioredoxin)-like domain；乌兰巴托:泛素;UPR：未折叠蛋白反应；VLP：病毒样颗粒；水疱性口炎病毒；WB: Western blotting；WT:野生型;XBP1: X-box结合蛋白
ABSTRACT Zaire ebolavirus (EBOV) causes a severe hemorrhagic fever in humans and non-human primates with high morbidity and mortality. EBOV infection is dependent on its structural glycoprotein (GP), but high levels of GP expression also trigger cell rounding, detachment, and downregulation of many surface molecules that is thought to contribute to its high pathogenicity. Thus, EBOV has evolved an RNA editing mechanism to reduce its GP expression and increase its fitness. We now report that the GP expression is also suppressed at the protein level in cells by protein disulfide isomerases (PDIs). Although PDIs promote oxidative protein folding by catalyzing correct disulfide formation in the endoplasmic reticulum (ER), PDIA3/ERp57 adversely triggered the GP misfolding by targeting GP cysteine residues and activated the unfolded protein response (UPR). Abnormally folded GP was targeted by ER-associated protein degradation (ERAD) machinery and, unexpectedly, was degraded via the macroautophagy/autophagy-lysosomal pathway, but not the proteasomal pathway. PDIA3 also decreased the GP expression from other ebolavirus species but increased the GP expression from Marburg virus (MARV), which is consistent with the observation that MARV-GP does not cause cell rounding and detachment, and MARV does not regulate its GP expression via RNA editing during infection. Furthermore, five other PDIs also had a similar inhibitory activity to EBOV-GP. Thus, PDIs negatively regulate ebolavirus glycoprotein expression, which balances the viral life cycle by maximizing their infection but minimizing their cellular effect. We suggest that ebolaviruses hijack the host protein folding and ERAD machinery to increase their fitness via reticulophagy during infection. Abbreviations: 3-MA: 3-methyladenine; 4-PBA: 4-phenylbutyrate; ACTB: β-actin; ATF: activating transcription factor; ATG: autophagy-related; BafA1: bafilomycin A1; BDBV: Bundibugyo ebolavirus; CALR: calreticulin; CANX: calnexin; CHX: cycloheximide; CMA: chaperone-mediated autophagy; ConA: concanamycin A; CRISPR: clusters of regularly interspaced short palindromic repeats; Cas9: CRISPR-associated protein 9; dsRNA: double-stranded RNA; EBOV: Zaire ebolavirus; EDEM: ER degradation enhancing alpha-mannosidase like protein; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; Env: envelope glycoprotein; ER: endoplasmic reticulum; ERAD: ER-associated protein degradation; ERN1/IRE1: endoplasmic reticulum to nucleus signaling 1; GP: glycoprotein; HA: hemagglutinin; HDAC6: histone deacetylase 6; HMM: high-molecular-mass; HIV-1: human immunodeficiency virus type 1; HSPA5/BiP: heat shock protein family A (Hsp70) member 5; IAV: influenza A virus; IP: immunoprecipitation; KIF: kifenesine; Lac: lactacystin; LAMP: lysosomal associated membrane protein; MAN1B1/ERManI: mannosidase alpha class 1B member 1; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MARV: Marburg virus; MLD: mucin-like domain; NHK/SERPINA1: alpha1-antitrypsin variant null (Hong Kong); NTZ: nitazoxanide; PDI: protein disulfide isomerase; RAVV: Ravn virus; RESTV: Reston ebolavirus; SARS-CoV: severe acute respiratory syndrome coronavirus; SBOV: Sudan ebolavirus; sGP: soluble GP; SQSTM1/p62: sequestosome 1; ssGP: small soluble GP; TAFV: Taï Forest ebolavirus; TIZ: tizoxanide; TGN: thapsigargin; TLD: TXN (thioredoxin)-like domain; Ub: ubiquitin; UPR: unfolded protein response; VLP: virus-like particle; VSV: vesicular stomatitis virus; WB: Western blotting; WT: wild-type; XBP1: X-box binding protein 1.