Proteomic identification of nuclear processes manipulated by cytomegalovirus early during infection.
Proteomic identification of nuclear processes manipulated by cytomegalovirus early during infection.
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DOI：
10.1002/pmic.201400599
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发表时间：
2015-06
期刊：
影响因子：
3.4
通讯作者：
Terhune SS
中科院分区：
文献类型：
--
作者：
Carter DM;Westdorp K;Noon KR;Terhune SS
人巨细胞病毒（HCMV）是一种疱疹病毒，广泛分布于世界各地，免疫抑制后可导致危及生命的疾病。HCMV表达许多蛋白质，其功能是建立支持病毒复制的细胞内环境。像大多数DNA病毒一样，HCMV操纵细胞核内的过程。我们已经定量的变化，在宿主细胞核蛋白质组感染后24 hpi与临床病毒分离。我们将SILAC与多个分馏阶段相结合来定义变化。在LTQ Orbitrap Velos质谱仪上通过RP-HPLC结合LC-MS/MS分析胰蛋白酶肽。用MaxQuant处理来自三个生物学重复的数据。共对1281种细胞蛋白质进行了定量，发现77种蛋白质存在显著差异表达。此外，我们还观察到36种与细胞核相关的病毒蛋白。多种生物学过程被显著改变，包括细胞周期、mRNA代谢和核质转运方面的增加，同时免疫应答降低。我们验证了几种蛋白质的变化，包括一个经典的核转运蛋白的子集。此外，我们证明了这些输入因子的破坏抑制HCMV复制。总的来说，我们已经确定了HCMV诱导的核蛋白质组变化，并发现了几个重要的感染过程。
Human cytomegalovirus (HCMV) is a herpesvirus that is ubiquitously distributed world-wide and causes life-threating disease upon immunosuppression. HCMV expresses numerous proteins that function to establish an intracellular environment that supports viral replication. Like most DNA viruses, HCMV manipulates processes within the nucleus. We have quantified changes in the host cell nuclear proteome at 24 hpi following infection with a clinical viral isolate. We have combined SILAC with multiple stages of fractionation to define changes. Tryptic peptides were analyzed by RP-HPLC combined with LC-MS/MS on an LTQ Orbitrap Velos mass spectrometer. Data from three biological replicates were processed with MaxQuant. A total of 1281 cellular proteins were quantified and 77 were found to be significantly differentially expressed. In addition, we observed 36 viral proteins associated with the nucleus. Diverse biological processes were significantly altered including increased aspects of cell cycling, mRNA metabolism, and nucleocytoplasmic transport while decreased immune responses. We validated changes for several proteins including a subset of classical nuclear transport proteins. In addition, we demonstrated that disruption of these import factors is inhibitory to HCMV replication. Overall, we have identified HCMV-induced changes in the nuclear proteome and uncovered several processes that are important for infection.