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Novel phages of Pseudomonas syringae unveil numerous potential auxiliary metabolic genes. 丁香假单胞菌的新型噬菌体揭示了大量潜在的辅助代谢基因。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.001990
Chloé Feltin, Julian R Garneau, Cindy E Morris, Annette Bérard, Clara Torres-Barceló

Relatively few phages that infect plant pathogens have been isolated and investigated. The Pseudomonas syringae species complex is present in various environments, including plants. It can cause major crop diseases, such as bacterial canker on apricot trees. This study presents a collection of 25 unique phage genomes that infect P. syringae. These phages were isolated from apricot orchards with bacterial canker symptoms after enrichment with 21 strains of P. syringae. This collection comprises mostly virulent phages, with only three being temperate. They belong to 14 genera, 11 of which are newly discovered, and 18 new species, revealing great genetic diversity within this collection. Novel DNA packaging systems have been identified bioinformatically in one of the new phage species, but experimental confirmation is required to define the precise mechanism. Additionally, many phage genomes contain numerous potential auxiliary metabolic genes with diversified putative functions. At least three phages encode genes involved in bacterial tellurite resistance, a toxic metalloid. This suggests that viruses could play a role in bacterial stress tolerance. This research emphasizes the significance of continuing the search for new phages in the agricultural ecosystem to unravel novel ecological diversity and new gene functions. This work contributes to the foundation for future fundamental and applied research on phages infecting phytopathogenic bacteria.

对感染植物病原体的噬菌体进行分离和研究的相对较少。丁香假单胞菌复合菌种存在于包括植物在内的各种环境中。它能引起主要的农作物病害,如杏树上的细菌性腐烂病。本研究收集了 25 个感染丁香假单胞菌的独特噬菌体基因组。这些噬菌体是用 21 株 P. syringae 菌株富集后从出现细菌性腐烂病症状的杏园中分离出来的。这些噬菌体主要是毒性噬菌体,其中只有 3 种是温性噬菌体。这些噬菌体属于 14 个属,其中 11 个属是新发现的,还有 18 个新种,揭示了这些噬菌体的遗传多样性。在其中一个新的噬菌体物种中,通过生物信息学方法发现了新的 DNA 包装系统,但要确定其确切的机制还需要实验证实。此外,许多噬菌体基因组包含许多潜在的辅助代谢基因,其推测功能多种多样。至少有三种噬菌体编码涉及细菌抗碲酸盐(一种有毒的类金属)的基因。这表明病毒可能在细菌耐受压力方面发挥作用。这项研究强调了在农业生态系统中继续寻找新噬菌体以揭示新的生态多样性和新的基因功能的重要性。这项工作为今后开展感染植物病原菌的噬菌体的基础研究和应用研究奠定了基础。
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引用次数: 0
Limited potexvirus diversity in eastern Gulf of Mexico seagrass meadows. 墨西哥湾东部海草草甸中有限的 potexvirus 多样性。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.002004
Shen Jean Lim, Karyna Rosario, Meredith E Kernbach, Anthony J Gross, Bradley T Furman, Mya Breitbart

Turtlegrass virus X, which infects the seagrass Thalassia testudinum, is the only potexvirus known to infect marine flowering plants. We investigated potexvirus distribution in seagrasses using a degenerate reverse transcription polymerase chain reaction (RT-PCR) assay originally designed to capture potexvirus diversity in terrestrial plants. The assay, which implements Potex-5 and Potex-2RC primers, successfully amplified a 584 nt RNA-dependent RNA polymerase (RdRp) fragment from TVX-infected seagrasses. Following validation, we screened 74 opportunistically collected, apparently healthy seagrass samples for potexviruses using this RT-PCR assay. The survey examined the host species T. testudinum, Halodule wrightii, Halophila stipulacea, Syringodium filiforme, Ruppia maritima, and Zostera marina. Potexvirus PCR products were successfully generated only from T. testudinum samples and phylogenetic analysis of sequenced PCR products revealed five distinct TVX sequence variants. Although the RT-PCR assay revealed limited potexvirus diversity in seagrasses, the expanded geographic distribution of TVX shown here emphasizes the importance of future studies to investigate T. testudinum populations across its native range and understand how the observed fine-scale genetic diversity affects host-virus interactions.

感染海草 Thalassia testudinum 的龟甲草病毒 X 是目前已知的唯一一种感染海洋开花植物的 potexvirus。我们使用一种退化的反转录聚合酶链反应(RT-PCR)检测方法调查了海草中的potex病毒分布情况,该检测方法最初是为捕捉陆生植物中的potex病毒多样性而设计的。该方法采用 Potex-5 和 Potex-2RC 引物,成功地从 TVX 感染的海草中扩增出 584 nt 的 RNA 依赖性 RNA 聚合酶(RdRp)片段。经过验证后,我们利用这种 RT-PCR 检测方法对 74 份随机采集的表面健康的海草样本进行了 potex 病毒筛查。调查对象包括宿主物种 T. testudinum、Halodule wrightii、Halophila stipulacea、Syringodium filiforme、Ruppia maritima 和 Zostera marina。仅从 T. testudinum 样品中成功生成了 Potexvirus PCR 产物,对已测序的 PCR 产物进行系统进化分析后发现了五个不同的 TVX 序列变体。尽管 RT-PCR 分析显示海草中的波坦病毒多样性有限,但本文所显示的 TVX 地理分布范围的扩大强调了未来研究的重要性,即调查 T. testudinum 在其原生地的种群,并了解所观察到的细粒度遗传多样性如何影响宿主与病毒之间的相互作用。
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引用次数: 0
Smallpox vaccination campaigns resulted in age-associated population cross-immunity against monkeypox virus. 天花疫苗接种活动导致了与年龄相关的人群对猴痘病毒的交叉免疫。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.001999
Kieran Dee, Maria Manali, Laura A Bissett, Jordan Bone, Callum Magill, Chris Davis, Brian J Willett, Pablo R Murcia

Increased human-to-human transmission of monkeypox virus (MPXV) is cause for concern, and antibodies directed against vaccinia virus (VACV) are known to confer cross-protection against Mpox. We used 430 serum samples derived from the Scottish patient population to investigate antibody-mediated cross-neutralization against MPXV. By combining electrochemiluminescence immunoassays with live-virus neutralization assays, we show that people born when smallpox vaccination was routinely offered in the United Kingdom have increased levels of antibodies that cross-neutralize MPXV. Our results suggest that age is a risk factor of Mpox infection, and people born after 1971 are at higher risk of infection upon exposure.

猴痘病毒(MPXV)在人与人之间传播的增加令人担忧,而针对疫苗病毒(VACV)的抗体可对猴痘病毒产生交叉保护作用。我们使用来自苏格兰患者群体的 430 份血清样本来研究抗体介导的针对 MPXV 的交叉中和作用。通过将电化学发光免疫测定与活病毒中和测定相结合,我们发现在英国常规提供天花疫苗接种时出生的人体内可交叉中和 MPXV 的抗体水平有所提高。我们的研究结果表明,年龄是天花病毒感染的一个风险因素,1971 年后出生的人接触天花病毒后感染的风险更高。
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引用次数: 0
H5N1 clade 2.3.4.4b avian influenza viruses replicate in differentiated bovine airway epithelial cells cultured at air-liquid interface. H5N1 2.3.4.4b 支禽流感病毒在空气-液体界面培养的分化牛气道上皮细胞中复制。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.002007
Luca Bordes, Nora M Gerhards, Stan Peters, Sophie van Oort, Marit Roose, Romy Dresken, Sandra Venema, Manouk Vrieling, Marc Engelsma, Wim H M van der Poel, Rik L de Swart

Highly pathogenic avian influenza (HPAI) H5N1 viruses are responsible for disease outbreaks in wild birds and poultry, resulting in devastating losses to the poultry sector. Since 2020, an increasing number of outbreaks of HPAI H5N1 was seen in wild birds. Infections in mammals have become more common, in most cases in carnivores after direct contact with infected birds. Although ruminants were previously not considered a host species for HPAI viruses, in March 2024 multiple outbreaks of HPAI H5N1 were detected in goats and cattle in the United States. Here, we have used primary bronchus-derived well-differentiated bovine airway epithelial cells (WD-AECs) cultured at air-liquid interface to assess the susceptibility and permissiveness of bovine epithelial cells to infection with European H5N1 virus isolates. We inoculated bovine WD-AECs with three low-passage HPAI clade 2.3.4.4b H5N1 virus isolates and detected rapid increases in viral genome loads and infectious virus during the first 24 h post-inoculation, without substantial cytopathogenic effects. Three days post-inoculation infected cells were still detectable by immunofluorescent staining. These data indicate that multiple lineages of HPAI H5N1 may have the propensity to infect the respiratory tract of cattle and support extension of avian influenza surveillance efforts to ruminants. Furthermore, this study underscores the benefit of WD-AEC cultures for pandemic preparedness by providing a rapid and animal-free assessment of the host range of an emerging pathogen.

高致病性禽流感 H5N1 病毒是野禽和家禽爆发疾病的罪魁祸首,给家禽业造成毁灭性损失。自 2020 年以来,野鸟中爆发 H5N1 型高致病性禽流感的数量不断增加。哺乳动物感染已变得越来越常见,大多数情况下是食肉动物直接接触受感染的鸟类后感染。虽然反刍动物以前不被认为是高致病性禽流感病毒的宿主物种,但 2024 年 3 月在美国的山羊和牛中发现了多起 H5N1 型高致病性禽流感疫情。在此,我们使用在空气-液体界面培养的原代支气管分化良好的牛气道上皮细胞(WD-AECs)来评估牛上皮细胞对欧洲 H5N1 病毒分离物感染的易感性和允许性。我们给牛 WD-AECs 接种了三种低传代高致病性禽流感 2.3.4.4b 支系 H5N1 病毒分离株,在接种后的头 24 小时内检测到病毒基因组载量和传染性病毒迅速增加,但没有产生实质性的细胞致病性影响。接种后三天,免疫荧光染色仍可检测到受感染的细胞。这些数据表明,高致病性禽流感 H5N1 的多个品系可能有感染牛呼吸道的倾向,因此支持将禽流感监测工作扩展到反刍动物。此外,这项研究还强调了 WD-AEC 培养物通过对新出现病原体的宿主范围进行快速、无动物干扰的评估,有利于大流行病的防备。
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引用次数: 0
Characterization of a natural 'dead-end' variant of Schmallenberg virus. 施马伦贝格病毒天然 "死胡同 "变种的特征。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.002005
Franziska Sick, Sophie Zeiske, Martin Beer, Kerstin Wernike

Schmallenberg virus (SBV) belongs to the Simbu serogroup within the family Peribunyaviridae, genus Orthobunyavirus and is transmitted by Culicoides biting midges. Infection of naïve ruminants in a critical phase of gestation may lead to severe congenital malformations. Sequence analysis from viremic animals revealed a very high genome stability. In contrast, sequence variations are frequently described for SBV from malformed fetuses. In addition to S segment mutations, especially within the M segment encoding the major immunogen Gc, point mutations or genomic deletions are also observed. Analysis of the SBV_D281/12 isolate from a malformed fetus revealed multiple point mutations in all three genome segments. It also has a large genomic deletion in the antigenic domain encoded by the M segment compared to the original SBV reference strain 'BH80/11' isolated from viremic blood in 2011. Interestingly, SBV_D281/12 showed a marked replication deficiency in vitro in Culicoides sonorensis cells (KC cells), but not in standard baby hamster kidney cells (BHK-21). We therefore generated a set of chimeric viruses of rSBV_D281/12 and wild-type rSBV_BH80/11 by reverse genetics, which were characterized in both KC and BHK-21 cells. It could be shown that the S segment of SBV_D281/12 is responsible for the replication deficit and that it acts independently from the large deletion within Gc. In addition, a single point mutation at position 111 (S to N) of the nucleoprotein was identified as the critical mutation. Our results suggest that virus variants found in malformed fetuses and carrying characteristic genomic mutations may have a clear 'loss of fitness' for their insect hosts in vitro. It can also be concluded that such mutations lead to virus variants that are no longer part of the natural transmission cycle between mammalian and insect hosts. Interestingly, analysis of a series of SBV sequences confirmed the S111N mutation exclusively in samples of malformed fetuses and not in blood from viremic animals. The characterization of these changes will allow the definition of protein functions that are critical for only one group of hosts.

施马伦贝格病毒(SBV)属于Peribunyaviridae科、Orthobunyavirus属的Simbu血清群,由咬蠓传播。天真反刍动物在妊娠的关键阶段受到感染可能会导致严重的先天性畸形。对病毒血症动物的序列分析表明,该病毒的基因组稳定性非常高。与此相反,畸形胎儿的 SBV 序列经常发生变异。除了 S 段突变(尤其是编码主要免疫原 Gc 的 M 段)外,还可观察到点突变或基因组缺失。对来自畸形胎儿的 SBV_D281/12 分离物的分析表明,该病毒的所有三个基因组片段都存在多个点突变。与 2011 年从病毒血症血液中分离出的原始 SBV 参考菌株 "BH80/11 "相比,它的 M 段编码的抗原结构域也有较大的基因组缺失。有趣的是,SBV_D281/12 在体外 Culicoides sonorensis 细胞(KC 细胞)中表现出明显的复制缺陷,而在标准小仓鼠肾细胞(BHK-21)中则没有。因此,我们通过反向遗传学方法产生了一组 rSBV_D281/12 和野生型 rSBV_BH80/11 的嵌合病毒,并在 KC 和 BHK-21 细胞中对其进行了鉴定。结果表明,SBV_D281/12 的 S 段是造成复制缺陷的原因,它的作用独立于 Gc 内的大缺失。此外,核蛋白第 111 位(S 到 N)的单点突变被确定为关键突变。我们的研究结果表明,在畸形胎儿体内发现的携带特征性基因组突变的病毒变异体在体外可能会对其昆虫宿主造成明显的 "健康损失"。还可以得出结论,这种突变导致病毒变种不再是哺乳动物和昆虫宿主之间自然传播循环的一部分。有趣的是,对一系列 SBV 序列的分析证实,S111N 变异只存在于畸形胎儿样本中,而不存在于有病毒感染的动物血液中。对这些变化进行定性分析,可以确定哪些蛋白质功能只对一类宿主至关重要。
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引用次数: 0
Whole-genome analysis of human group A rotaviruses in 1980s Japan and evolutionary assessment of global Wa-like strains across half a century. 20 世纪 80 年代日本人类 A 组轮状病毒的全基因组分析和半个世纪以来全球类似 Wa 病毒株的进化评估。
IF 3.8 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-06-01 DOI: 10.1099/jgv.0.001998
Yuya Fukuda, Kenji Kondo, Shuji Nakata, Yasuyuki Morita, Noriaki Adachi, Keiko Kogawa, Susumu Ukae, Yoshimasa Kudou, Shuhei Adachi, Masaki Yamamoto, Shinobu Fukumura, Takeshi Tsugawa

Historically, the Wa-like strains of human group A rotavirus (RVA) have been major causes of gastroenteritis. However, since the 2010s, the circulation of non-Wa-like strains has been increasingly reported, indicating a shift in the molecular epidemiology of RVA. Although understanding RVA evolution requires the analysis of both current and historical strains, comprehensive pre-1980's sequencing data are scarce globally. We determined the whole-genome sequences of representative strains from six RVA gastroenteritis outbreaks observed at an infant home in Sapporo, Japan, between 1981 and 1989. These outbreaks were mainly caused by G1 or G3 Wa-like strains, resembling strains from the United States in the 1970s-1980s and from Malawi in the 1990s. Phylogenetic analysis of these infant home strains, together with Wa-like strains collected worldwide from the 1970s to 2020, revealed a notable trend: pre-2010 strains diverged into multiple lineages in many genomic segments, whereas post-2010 strains tended to converge into a single lineage. However, Bayesian skyline plot indicated near-constant effective population sizes from the 1970s to 2020, and selection pressure analysis identified positive selection only at amino acid 75 of NSP2. These results suggest that evidence supporting the influence of rotavirus vaccines, introduced globally since 2006, on Wa-like RVA molecular evolution is lacking at present, and phylogenetic analysis may simply reflect natural fluctuations in RVA molecular evolution. Evaluating the long-term impact of RV vaccines on the molecular evolution of RVA requires sustained surveillance.

一直以来,人类 A 组轮状病毒(RVA)的类 Wa 型毒株是导致肠胃炎的主要原因。然而,自2010年代以来,非Wa样毒株流行的报道越来越多,这表明RVA的分子流行病学发生了变化。尽管了解 RVA 的进化需要对当前和历史菌株进行分析,但全球范围内 1980 年代以前的全面测序数据非常稀缺。我们测定了 1981 年至 1989 年间在日本札幌市一家婴儿之家观察到的六次 RVA 肠胃炎暴发中代表性菌株的全基因组序列。这些疫情主要由 G1 或 G3 类 Wa 菌株引起,类似于 20 世纪 70-80 年代美国和 20 世纪 90 年代马拉维的菌株。对这些婴幼儿家庭菌株以及 20 世纪 70 年代至 2020 年在全球收集的类 Wa 菌株进行的系统发育分析表明了一个明显的趋势:2010 年前的菌株在许多基因组片段上分化成多个系,而 2010 年后的菌株则趋向于汇聚成一个系。然而,贝叶斯天际线图显示,从 20 世纪 70 年代到 2020 年,有效种群大小近乎恒定,选择压力分析仅在 NSP2 的第 75 个氨基酸处发现了正选择。这些结果表明,目前还缺乏证据支持自 2006 年以来在全球范围内引入的轮状病毒疫苗对类似 Wa 的 RVA 分子进化的影响,系统发育分析可能只是反映了 RVA 分子进化的自然波动。评估 RV 疫苗对 RVA 分子进化的长期影响需要持续的监测。
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引用次数: 0
ICTV Virus Taxonomy Profile: Fimoviridae 2024 ICTV 病毒分类简介:Fimoviridae 2024
IF 3.8 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-02 DOI: 10.1099/jgv.0.001943
Michele Digiaro, Toufic Elbeaino, Kenji Kubota, Francisco M. Ochoa-Coron and Susanne von Bargen
Members of the family Fimoviridae are plant viruses with a multipartite negative-sense enveloped RNA genome (−ssRNA), composed of 4–10 segments comprising 12.3–18.5 kb in total, within quasi-spherical virions. Fimoviruses are transmitted to plants by eriophyid mites and induce characteristic cytopathologies in their host plants, including double membrane-bound bodies in the cytoplasm of virus-infected cells. Most fimoviruses infect dicotyledonous plants, and many cause serious disease epidemics. This is a summary of the ICTV Report on the family Fimoviridae, which is available at ictv.global/report/fimoviridae.
细小病毒科(Fimoviridae)的成员是植物病毒,其基因组(−ssRNA)由 4–10 个片段组成,总计 12.3–18.5 kb,位于准球形病毒体内。细小病毒通过螨虫传播给植物,并诱导宿主植物产生特有的细胞病理现象,包括病毒感染细胞的细胞质中出现双膜结合体。大多数细小病毒感染双子叶植物,其中许多会导致严重的疾病流行。本文是 ICTV 关于细小病毒科的报告摘要,可在 ictv.global/report/fimoviridae 上查阅。
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引用次数: 0
Iceland: an underestimated hub for the spread of high-pathogenicity avian influenza viruses in the North Atlantic 冰岛:被低估的北大西洋高致病性禽流感病毒传播中心
IF 3.8 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-02 DOI: 10.1099/jgv.0.001985
Ann Kathrin Ahrens, Stefán Ragnar Jónsson, Vilhjálmur Svansson, Brigitte Brugger, Martin Beer, Timm C. Harder and Anne Pohlmann
High-pathogenicity avian influenza viruses (HPAIVs) of the goose/Guangdong lineage are enzootically circulating in wild bird populations worldwide. This increases the risk of entry into poultry production and spill-over to mammalian species, including humans. Better understanding of the ecological and epizootiological networks of these viruses is essential to optimize mitigation measures. Based on full genome sequences of 26 HPAIV samples from Iceland, which were collected between spring and autumn 2022, as well as 1 sample from the 2023 summer period, we show that 3 different genotypes of HPAIV H5N1 clade 2.3.4.4b were circulating within the wild bird population in Iceland in 2022. Furthermore, in 2023 we observed a novel introduction of HPAIV H5N5 of the same clade to Iceland. The data support the role of Iceland as an utmost northwestern distribution area in Europe that might act also as a potential bridging point for intercontinental spread of HPAIV across the North Atlantic.
鹅/广东系高致病性禽流感病毒(HPAIV)在全球野鸟种群中流行。这增加了病毒进入家禽生产领域并蔓延到包括人类在内的哺乳动物物种的风险。更好地了解这些病毒的生态学和动物流行病学网络对于优化缓解措施至关重要。根据 2022 年春季至秋季期间从冰岛采集的 26 份 HPAIV 样本以及 2023 年夏季采集的 1 份样本的全基因组序列,我们发现 2022 年冰岛野鸟种群中流行着 3 种不同基因型的 HPAIV H5N1 支系 2.3.4.4b。此外,在 2023 年,我们还观察到同一支系的 HPAIV H5N5 新传入冰岛。这些数据支持冰岛作为欧洲最西北分布区的作用,它也可能成为高致病性禽流感跨北大西洋洲际传播的潜在桥梁。
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引用次数: 0
Erratum ICTV virus taxonomy profile: Fimoviridae 2024. 勘误 ICTV 病毒分类概况:Fimoviridae 2024。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-01 DOI: 10.1099/jgv.0.001992
Michele Digiaro, Toufic Elbeaino, Kenji Kubota, Francisco M Ochoa-Corona, Susanne von Bargen
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引用次数: 0
Japanese encephalitis virus hijacks ER-associated degradation regulators for its replication. 日本脑炎病毒劫持ER相关降解调节因子进行复制。
IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-05-01 DOI: 10.1099/jgv.0.001995
Riya Sarkar, Simran Chhabra, Mukesh Tanwar, Nisheeth Agarwal, Manjula Kalia

Flaviviruses target their replication on membranous structures derived from the ER, where both viral and host proteins play crucial structural and functional roles. Here, we have characterized the involvement of the ER-associated degradation (ERAD) pathway core E3 ligase complex (SEL1L-HRD1) regulator proteins in the replication of Japanese encephalitis virus (JEV). Through high-resolution immunofluorescence imaging of JEV-infected HeLa cells, we observe that the virus replication complexes marked by NS1 strongly colocalize with the ERAD adapter SEL1L, lectin OS9, ER-membrane shuttle factor HERPUD1, E3 ubiquitin ligase HRD1 and rhomboid superfamily member DERLIN1. NS5 positive structures also show strong overlap with SEL1L. While these effectors show significant transcriptional upregulation, their protein levels remain largely stable in infected cells. siRNA mediated depletion of OS9, SEL1L, HERPUD1 and HRD1 significantly inhibit viral RNA replication and titres, with SEL1L depletion showing the maximum attenuation of replication. By performing protein translation arrest experiments, we show that SEL1L, and OS9 are stabilised upon JEV infection. Overall results from this study suggest that these ERAD effector proteins are crucial host-factors for JEV replication.

黄病毒将其复制目标锁定在源自ER的膜结构上,病毒和宿主蛋白在其中发挥着关键的结构和功能作用。在这里,我们研究了ER相关降解(ERAD)途径核心E3连接酶复合物(SEL1L-HRD1)调节蛋白参与日本脑炎病毒(JEV)复制的特性。通过对感染 JEV 的 HeLa 细胞进行高分辨率免疫荧光成像,我们观察到以 NS1 为标志的病毒复制复合物与 ERAD 适配器 SEL1L、凝集素 OS9、ER-膜穿梭因子 HERPUD1、E3 泛素连接酶 HRD1 和斜方形超家族成员 DERLIN1 高度共定位。NS5 阳性结构与 SEL1L 也有很强的重叠。siRNA 介导的 OS9、SEL1L、HERPUD1 和 HRD1 基因耗竭能显著抑制病毒 RNA 复制和病毒滴度,其中 SEL1L 基因耗竭对病毒复制的抑制最大。通过进行蛋白翻译阻滞实验,我们发现 SEL1L 和 OS9 在感染 JEV 后趋于稳定。这项研究的总体结果表明,这些ERAD效应蛋白是JEV复制的关键宿主因子。
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Journal of General Virology
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