SARS-CoV-2 Is Restricted by Zinc Finger Antiviral Protein despite Preadaptation to the Low-CpG Environment in Humans.
SARS-CoV-2 Is Restricted by Zinc Finger Antiviral Protein despite Preadaptation to the Low-CpG Environment in Humans.
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DOI：
10.1128/mbio.01930-20
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发表时间：
2020-10-16
期刊：
影响因子：
6.4
通讯作者：
Kirchhoff F
中科院分区：
文献类型：
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作者：
Nchioua R;Kmiec D;Müller JA;Conzelmann C;Groß R;Swanson CM;Neil SJD;Stenger S;Sauter D;Münch J;Sparrer KMJ;Kirchhoff F
尽管干扰素抑制SARS-CoV-2，并已被评估用于治疗2019冠状病毒病（COVID-19），但最有效的类型和抗病毒效应物仍有待确定。在这里，我们表明，IFN-γ是特别有效的限制SARS-CoV-2和诱导表达的抗病毒因子ZAP在人肺细胞。敲除实验显示内源性ZAP显著限制SARS-CoV-2。我们进一步表明，ZAP特异性靶向的CpG二核苷酸在SARS-CoV-2基因组中受到强烈抑制，并且SARS-CoV-2的两个最接近的马蹄蝙蝠亲属显示出最低的基因组CpG含量。尽管如此，人类ZAP的短和长同种型都降低了SARS-CoV-2 RNA水平，并且这种活性在马蹄蝙蝠和穿山甲ZAP直系同源物中是保守的。我们的研究结果表明，II型干扰素对SARS-CoV-2特别有效，ZAP限制这种大流行性病毒病原体可能会促进针对COVID-19的有效免疫疗法的开发。最近的证据表明，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）是敏感的干扰素（IFN）。然而，最有效的干扰素类型和潜在的抗病毒效应物仍有待确定。在这里，我们表明，锌指抗病毒蛋白（ZAP），它优先靶向病毒RNA序列中的CpG二核苷酸，限制SARS-CoV-2。我们进一步证明ZAP及其辅因子KHNYN和TRIM 25在人肺细胞中表达。I型、II型和III型IFN均强烈抑制SARS-CoV-2并进一步诱导ZAP表达。综合序列分析显示，SARS-CoV-2及其来自马蹄蝙蝠的近亲分别在所有已知的人类和蝙蝠冠状病毒中表现出最强的CpG抑制。然而，内源性ZAP表达限制了SARS-CoV-2在人肺细胞中的复制，特别是在用IFN-α或IFN-γ处理后。人ZAP的长和短同种型都降低了SARS-CoV-2 RNA表达水平，但前者的效率更高。最后，我们表明，限制SARS-CoV-2的能力是保守的ZAP直系同源的水库蝙蝠和潜在的中间穿山甲宿主的人类冠状病毒。总之，我们的研究结果表明，ZAP是对SARS-CoV-2的先天性反应的重要效应子，尽管这种大流行性病原体是从预先适应人类低CpG环境的冠状病毒的人畜共患病中出现的。
Although interferons inhibit SARS-CoV-2 and have been evaluated for treatment of coronavirus disease 2019 (COVID-19), the most effective types and antiviral effectors remain to be defined. Here, we show that IFN-γ is particularly potent in restricting SARS-CoV-2 and in inducing expression of the antiviral factor ZAP in human lung cells. Knockdown experiments revealed that endogenous ZAP significantly restricts SARS-CoV-2. We further show that CpG dinucleotides which are specifically targeted by ZAP are strongly suppressed in the SARS-CoV-2 genome and that the two closest horseshoe bat relatives of SARS-CoV-2 show the lowest genomic CpG content of all coronavirus sequences available from this reservoir host. Nonetheless, both the short and long isoforms of human ZAP reduced SARS-CoV-2 RNA levels, and this activity was conserved in horseshoe bat and pangolin ZAP orthologues. Our findings indicating that type II interferon is particularly efficient against SARS-CoV-2 and that ZAP restricts this pandemic viral pathogen might promote the development of effective immune therapies against COVID-19. Recent evidence shows that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is sensitive to interferons (IFNs). However, the most effective types of IFNs and the underlying antiviral effectors remain to be defined. Here, we show that zinc finger antiviral protein (ZAP), which preferentially targets CpG dinucleotides in viral RNA sequences, restricts SARS-CoV-2. We further demonstrate that ZAP and its cofactors KHNYN and TRIM25 are expressed in human lung cells. Type I, II, and III IFNs all strongly inhibited SARS-CoV-2 and further induced ZAP expression. Comprehensive sequence analyses revealed that SARS-CoV-2 and its closest relatives from horseshoe bats showed the strongest CpG suppression among all known human and bat coronaviruses, respectively. Nevertheless, endogenous ZAP expression restricted SARS-CoV-2 replication in human lung cells, particularly upon treatment with IFN-α or IFN-γ. Both the long and the short isoforms of human ZAP reduced SARS-CoV-2 RNA expression levels, but the former did so with greater efficiency. Finally, we show that the ability to restrict SARS-CoV-2 is conserved in ZAP orthologues of the reservoir bat and potential intermediate pangolin hosts of human coronaviruses. Altogether, our results show that ZAP is an important effector of the innate response against SARS-CoV-2, although this pandemic pathogen emerged from zoonosis of a coronavirus that was preadapted to the low-CpG environment in humans.