Plasma membrane profiling defines an expanded class of cell surface proteins selectively targeted for degradation by HCMV US2 in cooperation with UL141.
Plasma membrane profiling defines an expanded class of cell surface proteins selectively targeted for degradation by HCMV US2 in cooperation with UL141.
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DOI：
10.1371/journal.ppat.1004811
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发表时间：
2015-04
期刊：
影响因子：
6.7
通讯作者：
Lehner PJ
中科院分区：
文献类型：
--
作者：
Hsu JL;van den Boomen DJ;Tomasec P;Weekes MP;Antrobus R;Stanton RJ;Ruckova E;Sugrue D;Wilkie GS;Davison AJ;Wilkinson GW;Lehner PJ
人类巨细胞病毒(HCMV)US2、US3、US6和US11协同作用，通过下调MHC-I类分子(MHC-I)，阻止CD8+T淋巴细胞对病毒感染细胞的免疫识别。在这里，我们展示了US2的功能远远超出了MHC-I的退化。利用质膜图谱的系统蛋白质组学研究表明，Us2在下调其他细胞靶点方面是独一无二的，包括：5个不同的整合素α链、CD112、白细胞介素12受体、PTPRJ和血栓调节蛋白。US2招募了细胞E3连接酶TRC8来指导其所有靶标的蛋白酶体降解，这让人想起它对MHC-I的降解。而整合素α链被选择性降解，其整合素β1结合伙伴在内质网中积累。因此，整合素信号转导、细胞黏附和迁移受到强烈抑制。US2是降解大多数底物的必要条件和充分条件，但值得注意的是，HCMV NK细胞逃避功能UL141需要US2来加强NK细胞配体CD112的下调。UL141将CD112保留在内质网中，US2从内质网促进其依赖TRC8的逆转位和降解。这些发现将US2重新定义为一个多功能的降解中心，通过募集细胞E3连接酶TRC8，调节参与抗原提呈、NK细胞激活、迁移和凝血的多种免疫途径；并强调US2在巨细胞病毒致病中的S影响。作为最大的人类疱疹病毒，HCMV是病毒免疫逃避的典范，并进化出多种机制来逃避免疫检测并使其能够存活。HCMV基因US2、US3、US6和US11通过下调细胞表面MHC来促进病毒的持久性。我们开发了“质膜图谱”(PMP)，这是一种基于SILAC的无偏倚蛋白质组学技术，目的是询问MHC分子是否是这些基因的唯一焦点，或者是否其他细胞免疫受体也是目标。PMP比较对照细胞和病毒基因表达细胞的细胞表面受体的相对丰度。我们发现，虽然US3、US6和US11具有显著的MHC特异性，但US2可调节多种细胞表面免疫受体的表达。Us2介导的整合素α链蛋白酶体降解阻断了整合素信号转导，抑制了细胞的黏附和迁移。所有的US2底物都是通过细胞E3连接酶TRC8降解的，在一个值得注意的HCMV免疫-Easins之间协同作用的例子中，UL141要求US2靶向NK细胞配体CD112进行蛋白酶体降解。因此，HCMV US2和UL141是多种免疫相关途径的调节器，并作为一个多功能的降解中心，抑制HCMV感染细胞的迁移、免疫识别和杀伤。
Human cytomegalovirus (HCMV) US2, US3, US6 and US11 act in concert to prevent immune recognition of virally infected cells by CD8+ T-lymphocytes through downregulation of MHC class I molecules (MHC-I). Here we show that US2 function goes far beyond MHC-I degradation. A systematic proteomic study using Plasma Membrane Profiling revealed US2 was unique in downregulating additional cellular targets, including: five distinct integrin α-chains, CD112, the interleukin-12 receptor, PTPRJ and thrombomodulin. US2 recruited the cellular E3 ligase TRC8 to direct the proteasomal degradation of all its targets, reminiscent of its degradation of MHC-I. Whereas integrin α-chains were selectively degraded, their integrin β1 binding partner accumulated in the ER. Consequently integrin signaling, cell adhesion and migration were strongly suppressed. US2 was necessary and sufficient for degradation of the majority of its substrates, but remarkably, the HCMV NK cell evasion function UL141 requisitioned US2 to enhance downregulation of the NK cell ligand CD112. UL141 retained CD112 in the ER from where US2 promoted its TRC8-dependent retrotranslocation and degradation. These findings redefine US2 as a multifunctional degradation hub which, through recruitment of the cellular E3 ligase TRC8, modulates diverse immune pathways involved in antigen presentation, NK cell activation, migration and coagulation; and highlight US2’s impact on HCMV pathogenesis. As the largest human herpesvirus, HCMV is a paradigm of viral immune evasion and has evolved multiple mechanisms to evade immune detection and enable survival. The HCMV genes US2, US3, US6 and US11 promote virus persistence by their ability to downregulate cell surface MHC. We developed ‘Plasma Membrane Profiling’ (PMP), an unbiased SILAC-based proteomics technique to ask whether MHC molecules are the only focus of these genes, or whether additional cellular immunoreceptors are also targeted. PMP compares the relative abundance of cell surface receptors between control and viral gene expressing cells. We found that whereas US3, US6 and US11 were remarkably MHC specific, US2 modulated expression of a wide variety of cell surface immunoreceptors. US2-mediated proteasomal degradation of integrin α-chains blocked integrin signaling and suppressed cell adhesion and migration. All US2 substrates were degraded via the cellular E3 ligase TRC8, and in a remarkable example of cooperativity between HCMV immune-evasins, UL141 requisitioned US2 to target the NK cell ligand CD112 for proteasomal degradation. HCMV US2 and UL141 are therefore modulators of multiple immune-related pathways and act as a multifunctional degradation hub that inhibits the migration, immune recognition and killing of HCMV-infected cells.
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影响因子：
64.8
作者：
Lilley, BN;Ploegh, HL
通讯作者：
Ploegh, HL
影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
32.4
作者：
FAHNESTOCK, ML;JOHNSON, JL;BJORKMAN, PJ
通讯作者：
BJORKMAN, PJ
DOI：
10.1084/jem.20092140
发表时间：
2010-06-07
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Jongbloed SL;Kassianos AJ;McDonald KJ;Clark GJ;Ju X;Angel CE;Chen CJ;Dunbar PR;Wadley RB;Jeet V;Vulink AJ;Hart DN;Radford KJ
通讯作者：
Radford KJ
影响因子：
3.8
作者：
Dolan, A;Cunningham, C;Davison, AJ
通讯作者：
Davison, AJ