Comparative Proteomics Reveals Strain-Specific β-TrCP Degradation via Rotavirus NSP1 Hijacking a Host Cullin-3-Rbx1 Complex.
Comparative Proteomics Reveals Strain-Specific β-TrCP Degradation via Rotavirus NSP1 Hijacking a Host Cullin-3-Rbx1 Complex.
复制标题
DOI：
10.1371/journal.ppat.1005929
复制
发表时间：
2016-10
期刊：
影响因子：
6.7
通讯作者：
Greenberg HB
中科院分区：
文献类型：
--
作者：
Ding S;Mooney N;Li B;Kelly MR;Feng N;Loktev AV;Sen A;Patton JT;Jackson PK;Greenberg HB
轮状病毒（RV）是幼儿严重胃肠炎的主要原因，每年在全世界造成50万人死亡。RV编码非结构蛋白1（NSP 1），其是一种充分表征的干扰素（IFN）拮抗剂，其通过介导宿主抗病毒因子（包括IRF 3和β-TrCP）的降解来促进病毒复制。在这里，我们使用六种人类和动物RV NSP 1作为诱饵，并进行串联亲和纯化，结合高分辨率质谱法来全面表征NSP 1-宿主蛋白相互作用网络。鉴定了多个Cullin-RING泛素连接酶（CRL）复合物。重要的是，通过siRNA沉默或化学扰动抑制cullin-3（Cul 3）或RING盒蛋白1（Rbx 1），可显著损害菌株特异性NSP 1介导的β-TrCP降解。从机制上讲，我们证明了NSP 1与宿主Cul 3-Rbx 1 CRL复合物一起定位于高尔基体，该复合物靶向β-TrCP和NSP 1以在蛋白酶体处共同破坏。我们的研究揭示了RV用于促进β-TrCP周转的新机制，并为病毒介导的先天免疫抑制提供了分子见解。轮状病毒（RV）是幼儿腹泻的主要原因，每年导致全球超过215，000人死亡。对于几乎每种哺乳动物，都有特别适合在宿主物种中有效复制的RV菌株。RV的成功在很大程度上源于其以宿主范围受限的方式抑制宿主抗病毒反应的非凡能力。一种名为NSP 1的病毒蛋白主要负责这一过程。在本文中，我们使用无偏的方法发现，NSP 1通过促进β-TrCP降解来抑制宿主先天免疫应答的能力取决于其与RV感染细胞内称为Cullin-E3连接酶复合物的蛋白质降解机器的特异性相互作用。阻断这种复合物极大地降低了NSP 1拮抗宿主免疫应答的能力，并且对RV复制有害。我们的研究揭示了Cullin-E3复合物在RV免疫逃避中的意想不到的作用，并对其他病毒病原体具有广泛的意义。
Rotaviruses (RVs) are the leading cause of severe gastroenteritis in young children, accounting for half a million deaths annually worldwide. RV encodes non-structural protein 1 (NSP1), a well-characterized interferon (IFN) antagonist, which facilitates virus replication by mediating the degradation of host antiviral factors including IRF3 and β-TrCP. Here, we utilized six human and animal RV NSP1s as baits and performed tandem-affinity purification coupled with high-resolution mass spectrometry to comprehensively characterize NSP1-host protein interaction network. Multiple Cullin-RING ubiquitin ligase (CRL) complexes were identified. Importantly, inhibition of cullin-3 (Cul3) or RING-box protein 1 (Rbx1), by siRNA silencing or chemical perturbation, significantly impairs strain-specific NSP1-mediated β-TrCP degradation. Mechanistically, we demonstrate that NSP1 localizes to the Golgi with the host Cul3-Rbx1 CRL complex, which targets β-TrCP and NSP1 for co-destruction at the proteasome. Our study uncovers a novel mechanism that RV employs to promote β-TrCP turnover and provides molecular insights into virus-mediated innate immunity inhibition. Rotaviruses (RVs) are the leading cause of diarrhea in young children and lead to over 215,000 deaths annually worldwide. For virtually every mammal, there are RV strains specifically adapted to replicate efficiently in that host species. The success of RV, to a large extent, derives from its extraordinary ability to suppress the host antiviral responses in a host range restricted manner. One viral protein, named NSP1, is primarily responsible for this process. In this paper, using an unbiased approach, we discovered that NSP1’s ability to inhibit the host innate immune responses by promoting β-TrCP degradation is dependent on its specific interaction with a protein degradation machine called the Cullin-E3 ligase complex within the RV infected cell. Blocking this complex tremendously reduces NSP1’s ability to antagonize the host immune response and is detrimental for RV replication. Our study uncovers an unexpected role of Cullin-E3 complex in RV immune evasion and has broad implications for other viral pathogens.
登录
查看更多内容
影响因子：
16
作者：
Davis, Zoe H.;Verschueren, Erik;Jang, Gwendolyn M.;Kleffman, Kevin;Johnson, Jeffrey R.;Park, Jimin;Von Dollen, John;Maher, M. Cyrus;Johnson, Tasha;Newton, William;Jaeger, Stefanie;Shales, Michael;Horner, Julie;Hernandez, Ryan D.;Krogan, Nevan J.;Glaunsinger, Britt A.
通讯作者：
Glaunsinger, Britt A.
影响因子：
13.5
作者：
Bolen, Christopher R.;Ding, Siyuan;Robek, Michael D.;Kleinstein, Steven H.
通讯作者：
Kleinstein, Steven H.
影响因子：
5.3
作者：
Cullinan, SB;Gordan, JD;Diehl, JA
通讯作者：
Diehl, JA
影响因子：
5.4
作者：
Barro, Mario;Patton, John T.
通讯作者：
Patton, John T.
影响因子：
6.4
作者：
HOSHINO, Y;WYATT, RG;KAPIKIAN, AZ
通讯作者：
KAPIKIAN, AZ