Functional Identification and Characterization of the Nuclear Egress Complex of a Gammaherpesvirus
Functional Identification and Characterization of the Nuclear Egress Complex of a Gammaherpesvirus
复制标题
伽马疱疹病毒核出口复合体的功能鉴定和表征
DOI：
10.1128/jvi.01422-19
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发表时间：
2019-09
影响因子：
5.4
通讯作者：
Deng Hongyu
中科院分区：
文献类型：
--
作者：
Lv Ying;Shen Sheng;Xiang Lingjiao;Jia Xing;Hou Yanjie;Wang Dacheng;Deng Hongyu
越来越多的知识表明，核出口复合物（NEC）对于疱疹病毒衣壳的核出口至关重要，可以将其视为囊泡介导的穿过核膜的运输途径。在这项研究中，我们确定了开放阅读框 67 (ORF67) 和 ORF69 是鼠伽马疱疹病毒 68 (MHV-68) 中 NEC 的组成部分，并证明它们能够有效地从哺乳动物细胞的核膜诱导病毒体样囊泡。这是首次发现伽马疱疹病毒的 NEC 表现出如此重要的特征。此外，我们还确定了对于介导 ORF67 和 ORF69 之间的相互作用以及核排出至关重要的氨基酸。值得注意的是，这些氨基酸在卡波西肉瘤相关疱疹病毒（KSHV）和EB病毒（EBV）中是保守的，为设计抗γ疱疹病毒药物提供了结构基础。摘要 疱疹病毒核出口复合物（NEC）由两种病毒蛋白组成。它们通过促进衣壳出芽进入核周空间（PNS），在介导衣壳从细胞核到细胞质的易位中发挥关键作用。在没有其他病毒蛋白的情况下，α疱疹病毒的NEC可以诱导从核膜形成病毒颗粒样囊泡。然而，γ疱疹病毒的NEC是否具有在哺乳动物细胞中这样做的能力仍有待确定。在这项研究中，我们首先构建了鼠伽马疱疹病毒 68 (MHV-68) 的开放阅读框 67 (ORF67) 缺失和 ORF69 缺失突变体，并证明 ORF67 和 ORF69 在核出口以及病毒裂解复制中发挥着关键作用。生化和生物成像分析表明，ORF67和ORF69相互作用，足以诱导哺乳动物细胞核膜形成病毒颗粒样囊泡。因此，我们指定 MHV-68 NEC 的 ORF67 和 ORF69 组件。此外，我们通过同源模型确定了介导 ORF67 和 ORF69 之间相互作用的关键氨基酸，并验证了它们在核出口中的功能，为伽玛疱疹病毒中 NEC 形成的分子基础提供了见解。重要性越来越多的知识表明，核出口复合物（NEC）对于疱疹病毒衣壳的核出口至关重要，可以将其视为通过核膜的囊泡介导的运输途径。在这项研究中，我们确定了开放阅读框 67 (ORF67) 和 ORF69 是鼠伽马疱疹病毒 68 (MHV-68) 中 NEC 的组成部分，并证明它们能够有效地从哺乳动物细胞的核膜诱导病毒体样囊泡。这是首次发现伽马疱疹病毒的 NEC 表现出如此重要的特征。此外，我们还确定了对于介导 ORF67 和 ORF69 之间的相互作用以及核排出至关重要的氨基酸。值得注意的是，这些氨基酸在卡波西肉瘤相关疱疹病毒（KSHV）和EB病毒（EBV）中是保守的，为设计抗γ疱疹病毒药物提供了结构基础。
Increasing amounts of knowledge indicate that the nuclear egress complex (NEC) is critical for the nuclear egress of herpesvirus capsids, which can be viewed as a vesicle-mediated transport pathway through the nuclear membrane. In this study, we identified open reading frame 67 (ORF67) and ORF69 as components of the NEC in murine gammaherpesvirus 68 (MHV-68) and demonstrated that they efficiently induce virion-like vesicles from the nuclear membrane in mammalian cells. This is the first time that the NEC of a gammaherpesvirus has been found to demonstrate such an essential characteristic. In addition, we identified amino acids critical for mediating the interaction between ORF67 and ORF69 as well as nuclear egress. Notably, these amino acids are conserved in Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV), providing a structural basis to design antigammaherpesvirus drugs. ABSTRACT The herpesvirus nuclear egress complex (NEC) is composed of two viral proteins. They play key roles in mediating the translocation of capsids from the nucleus to the cytoplasm by facilitating the budding of capsids into the perinuclear space (PNS). The NEC of alphaherpesvirus can induce the formation of virion-like vesicles from the nuclear membrane in the absence of other viral proteins. However, whether the NEC of gammaherpesvirus harbors the ability to do so in mammalian cells remains to be determined. In this study, we first constructed open reading frame 67 (ORF67)-null and ORF69-null mutants of murine gammaherpesvirus 68 (MHV-68) and demonstrated that both ORF67 and ORF69 play critical roles in nuclear egress and hence viral lytic replication. Biochemical and bioimaging analyses showed that ORF67 and ORF69 interacted with each other and were sufficient to induce the formation of virion-like vesicles from the nuclear membrane in mammalian cells. Thus, we designated ORF67 and ORF69 components of MHV-68 NEC. Furthermore, we identified amino acids critical for mediating the interaction between ORF67 and ORF69 through homology modeling and verified their function in nuclear egress, providing insights into the molecular basis of NEC formation in gammaherpesviruses. IMPORTANCE Increasing amounts of knowledge indicate that the nuclear egress complex (NEC) is critical for the nuclear egress of herpesvirus capsids, which can be viewed as a vesicle-mediated transport pathway through the nuclear membrane. In this study, we identified open reading frame 67 (ORF67) and ORF69 as components of the NEC in murine gammaherpesvirus 68 (MHV-68) and demonstrated that they efficiently induce virion-like vesicles from the nuclear membrane in mammalian cells. This is the first time that the NEC of a gammaherpesvirus has been found to demonstrate such an essential characteristic. In addition, we identified amino acids critical for mediating the interaction between ORF67 and ORF69 as well as nuclear egress. Notably, these amino acids are conserved in Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein-Barr virus (EBV), providing a structural basis to design antigammaherpesvirus drugs.
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影响因子：
6.7
作者：
Lee S;Salwinski L;Zhang C;Chu D;Sampankanpanich C;Reyes NA;Vangeloff A;Xing F;Li X;Wu TT;Sahasrabudhe S;Deng H;Lacount DJ;Sun R
通讯作者：
Sun R
影响因子：
5
作者：
K. Schulz;B. Klupp;H. Granzow;L. Passvogel;T. Mettenleiter
通讯作者：
K. Schulz;B. Klupp;H. Granzow;L. Passvogel;T. Mettenleiter
影响因子：
11.4
作者：
Bigalke, Janna M.;Heldwein, Ekaterina E.
通讯作者：
Heldwein, Ekaterina E.
影响因子：
5.4
作者：
Gonnella, R;Farina, A;Faggioni, A
通讯作者：
Faggioni, A
DOI：
10.1073/pnas.0701757104
发表时间：
2007-04-24
影响因子：
11.1
作者：
Klupp, Barbara G.;Granzow, Harald;Mettenleiter, Thomas C.
通讯作者：
Mettenleiter, Thomas C.