Recovery of paramyxovirus simian virus 5 with a V protein lacking the conserved cysteine-rich domain:: The multifunctional V protein blocks both interferon-β induction and interferon signaling
Recovery of paramyxovirus simian virus 5 with a V protein lacking the conserved cysteine-rich domain:: The multifunctional V protein blocks both interferon-β induction and interferon signaling
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DOI：
10.1006/viro.2002.1738
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发表时间：
2002-11-10
期刊：
影响因子：
3.7
通讯作者：
Lamb, RA
中科院分区：
文献类型：
--
作者：
He, B;Paterson, RG;Lamb, RA
副粘病毒猿猴病毒5（SV 5）的V蛋白是一种多功能蛋白，其含有与P蛋白共享的N-末端164个残基结构域和富含半胱氨酸且在副粘病毒中高度保守的独特C-末端结构域。我们报告从Vero细胞[干扰素（IFN）nonproducing细胞]的重组SV 5（rSV 5），缺乏V蛋白C-末端特异性结构域（rSV 5VDeltaC）的回收。在Vero细胞中，rSV 5VDeltaC形成大噬斑，并以与rSV 5相似的速率和滴度生长。在BHK或CV-1细胞中，rSV 5VDeltaC形成小斑块且生长不良。然而，即使在Vero细胞中生长，rSV 5VDeltaC在4至5代中也恢复为假野生型病毒，这表明V蛋白对于SV 5的成功复制的重要性。尽管rSV 5在许多细胞类型中生长，具有最小的细胞病变效应（CPE），但rSV 5VDeltaC在相同的细胞类型中引起广泛的CPE。为了克服由IFN诱导的抗病毒状态，许多病毒已经进化出通过阻断IFN的产生和消除IFN信号传导来抵消IFN的作用的机制。rSV 5通过介导STAT 1的降解阻断IFN信号传导，而rSV 5VDeltaC不引起STAT 1的降解，并且IFN信号传导通过ISGF 3转录复合物的形成而发生。此外，我们发现rSV 5感染细胞会阻止IFN-β的产生。IFN-β基因转录所需的转录因子IRF-3在rSV 5感染的细胞中不从细胞质易位到细胞核。相反，在rSV 5VDeltaC感染的细胞中，IRF-3主要定位于细胞核中，并产生IFN-β。通过使用IRF-3的异位表达，表明在dsRNA处理和V蛋白的表达后，IRF-3保留在细胞质中，而在dsRNA处理和P蛋白（其缺乏C-末端富含半胱氨酸的结构域）的表达后，IRF-3主要定位在细胞核中。因此，SV 5阻断先天免疫应答的两个不同途径，这两个途径都需要多功能SV 5 V蛋白的C末端特异性富含半胱氨酸的结构域的存在。(C)2002 Elsevier Science（美国）。
The V protein of the Paramyxovirus simian virus 5 (SV5) is a multifunctional protein containing an N-terminal 164 residue domain that is shared with the P protein and a distinct C-terminal domain that is cysteine-rich and which is highly conserved among Paramyxoviruses. We report the recovery from Vero cells [interferon (IFN) nonproducing cells] of a recombinant SV5 (rSV5) that lacks the V protein C-terminal specific domain (rSV5VDeltaC). In Vero cells rSV5VDeltaC forms large plaques and grows at a rate and titer similar to those of rSV5. In BHK or CV-1 cells rSV5VDeltaC forms small plaques and grows poorly. However, even when grown in Vero cells rSV5VDeltaC reverts to pseudo-wild-type virus in four to five passages, indicating the importance of the V protein for successful replication of SV5. Whereas rSV5 grows in many cell types with minimal cytopathic effect (CPE), rSV5VDeltaC causes extensive CPE in the same cell types. To overcome the antiviral state induced by IFN, many viruses have evolved mechanisms to counteract the effects of IFN by blocking the production of IFN and abrogating IFN signaling. Whereas rSV5 blocks IFN signaling by mediating the degradation of STAT1, rSV5VDeltaC does not cause the degradation of STAT1 and IFN signaling occurs through formation of the ISGF3 transcription complex. Furthermore, we find that rSV5 infection of cells prevents production of IFN-beta. The transcription factor IRF-3 which is required for transcription of the IFN-beta gene is not translocated from the cytoplasm to the nucleus in rSV5-infected cells. In contrast, in rSV5VDeltaC-infected cells IRF-3 is localized predominantly in the nucleus and IFN-beta is produced. By using ectopic expression of IRF-3, it was shown that after dsRNA treatment and expression of the V protein IRF-3 remained in the cytoplasm, whereas after dsRNA treatment and expression of the P protein (which lacks the C-terminal cysteine-rich domain) IRF-3 was localized predominantly in-the nucleus. Thus, SV5 blocks two distinct pathways of the innate immune response, both of which require the presence of the C-terminal specific cysteine-rich domain of the multifunctional SV5 V protein. (C) 2002 Elsevier Science (USA).