Porcine RING Finger Protein 114 Inhibits Classical Swine Fever Virus Replication via K27-Linked Polyubiquitination of Viral NS4B
Porcine RING Finger Protein 114 Inhibits Classical Swine Fever Virus Replication via K27-Linked Polyubiquitination of Viral NS4B
复制标题
猪环指蛋白114通过病毒NS4B的K27连接多泛素化抑制猪瘟病毒复制
DOI：
10.1128/jvi.01248-19
复制
发表时间：
2019-11-01
影响因子：
5.4
通讯作者：
Qiu, Hua-Ji
中科院分区：
文献类型：
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作者：
Zhang, Yuexiu;Zhang, Huawei;Qiu, Hua-Ji
猪RING指蛋白114（pRNF 114）是RING结构域E3连接酶的成员。本研究表明，pRNF 114是一种潜在的抗猪瘟病毒因子，其抗猪瘟病毒作用依赖于其E3连接酶活性。值得注意的是，pRNF 114靶向并催化NS 4 B蛋白的K27连接的多泛素化，然后促进NS 4 B的蛋白酶体依赖性降解，抑制CSFV的复制。据我们所知，pRNF 114是第一个被鉴定为参与抗CSFV活性的E3连接酶，靶向NS 4 B可能是抗病毒开发的关键途径。在宿主中，许多RING结构域E3连接酶被报道通过各种机制抑制病毒复制。在先前的筛选中，我们发现猪RING指蛋白114（pRNF 114），一个RING结构域E3泛素连接酶，抑制猪瘟病毒（CSFV）的复制。本研究旨在阐明pRNF 114抗猪瘟病毒的机制。CSFV感染后，pRNF 114 mRNA在体外和体内均上调。通过慢病毒递送系统在稳定表达pRNF 114的PK-pRNF 114细胞中显著抑制CSFV复制，而通过CRISPR/Cas9系统在RNF 114敲除的PK-15细胞中增强CSFV生长。具有E3泛素连接酶活性的pRNF 114的RING结构域对其抗病毒活性至关重要。pRNF 114通过其C-末端结构域与CSFV NS 4 B蛋白相互作用，这导致K27连接的多聚泛素化，并通过蛋白酶体依赖性途径降解NS 4 B。总的来说，这些发现表明pRNF 114作为CSFV复制的关键调节剂，并揭示了pRNF 114利用其E3泛素连接酶活性抑制CSFV复制的机制。猪RING指蛋白114（pRNF 114）是RING结构域E3连接酶的成员。本研究表明，pRNF 114是一种潜在的抗猪瘟病毒因子，其抗猪瘟病毒作用依赖于其E3连接酶活性。值得注意的是，pRNF 114靶向并催化NS 4 B蛋白的K27连接的多泛素化，然后促进NS 4 B的蛋白酶体依赖性降解，抑制CSFV的复制。据我们所知，pRNF 114是第一个被鉴定为参与抗CSFV活性的E3连接酶，靶向NS 4 B可能是抗病毒开发的关键途径。
Porcine RING finger protein 114 (pRNF114) is a member of the RING domain E3 ligases. In this study, it was shown that pRNF114 is a potential anti-CSFV factor and the anti-CSFV effect of pRNF114 depends on its E3 ligase activity. Notably, pRNF114 targets and catalyzes the K27-linked polyubiquitination of the NS4B protein and then promotes proteasome-dependent degradation of NS4B, inhibiting the replication of CSFV. To our knowledge, pRNF114 is the first E3 ligase to be identified as being involved in anti-CSFV activity, and targeting NS4B could be a crucial route for antiviral development. ABSTRACT In the host, many RING domain E3 ligases have been reported to inhibit viral replication through various mechanisms. In a previous screen, we found that porcine RING finger protein 114 (pRNF114), a RING domain E3 ubiquitin ligase, inhibits classical swine fever virus (CSFV) replication. This study aimed to clarify the underlying antiviral mechanism of pRNF114 against CSFV. Upon CSFV infection, pRNF114 mRNA was upregulated both in vitro and in vivo. CSFV replication was significantly suppressed in PK-pRNF114 cells stably expressing pRNF114 by the lentivirus-delivered system, whereas CSFV growth was enhanced in PK-15 cells with RNF114 knockout by the CRISPR/Cas9 system. The RING domain of pRNF114, which has E3 ubiquitin ligase activity, is crucial for its antiviral activity. Mechanistically, pRNF114 interacted with the CSFV NS4B protein through their C-terminal domains, which led to the K27-linked polyubiquitination and degradation of NS4B through a proteasome-dependent pathway. Collectively, these findings indicate that pRNF114 as a critical regulator of CSFV replication and uncover a mechanism by which pRNF114 employs its E3 ubiquitin ligase activity to inhibit CSFV replication. IMPORTANCE Porcine RING finger protein 114 (pRNF114) is a member of the RING domain E3 ligases. In this study, it was shown that pRNF114 is a potential anti-CSFV factor and the anti-CSFV effect of pRNF114 depends on its E3 ligase activity. Notably, pRNF114 targets and catalyzes the K27-linked polyubiquitination of the NS4B protein and then promotes proteasome-dependent degradation of NS4B, inhibiting the replication of CSFV. To our knowledge, pRNF114 is the first E3 ligase to be identified as being involved in anti-CSFV activity, and targeting NS4B could be a crucial route for antiviral development.