Ribosome Profiling Reveals Translational Upregulation of Cellular Oxidative Phosphorylation mRNAs during Vaccinia Virus-Induced Host Shutoff
Ribosome Profiling Reveals Translational Upregulation of Cellular Oxidative Phosphorylation mRNAs during Vaccinia Virus-Induced Host Shutoff
复制标题
核糖体分析揭示痘苗病毒诱导的宿主关闭期间细胞氧化磷酸化 mRNA 的翻译上调
DOI：
10.1128/jvi.01858-16
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发表时间：
2017-03-01
影响因子：
5.4
通讯作者：
Yang, Zhilong
中科院分区：
文献类型：
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作者：
Dai, Aimei;Cao, Shuai;Yang, Zhilong
摘要牛痘病毒感染引起宿主关闭，其特征在于宿主蛋白质合成的全面抑制。虽然宿主关闭可能有助于重新分配细胞资源用于病毒复制和逃避宿主抗病毒免疫应答，但它对病毒复制重要的细胞蛋白的连续合成构成挑战。然而，目前还不清楚某些细胞蛋白质是否以及如何在牛痘病毒诱导的宿主关闭期间选择性地合成。使用同步RNA测序和核糖体分析，两种技术定量全基因组水平的mRNA和活性蛋白质翻译，分别，我们分析了宿主细胞的反应，牛痘病毒感染的转录和翻译水平。分析表明，细胞mRNA缺失在宿主蛋白质合成的关闭中起主导作用。虽然细胞mRNA显著减少，但一部分细胞mRNA的相对翻译效率增加，特别是那些参与氧化磷酸化的mRNA，它们负责细胞能量的产生。进一步的实验表明，蛋白质水平和氧化磷酸化的活性增加，在牛痘病毒感染，而细胞氧化磷酸化功能的抑制显着抑制牛痘病毒的复制。氧化磷酸化mRNA的5′端非翻译区较短，对氧化磷酸化mRNA的翻译上调有贡献。这些结果提供了一种机制，耦合翻译控制和能量代谢，两个过程，所有病毒依赖于宿主细胞提供，以支持牛痘病毒复制在主机关闭的证据。重要性许多病毒感染导致全球宿主蛋白质合成关闭。虽然宿主蛋白质合成关闭通过将细胞资源重新定位到病毒复制而使病毒受益，但如果需要连续的蛋白质合成，则它也对维持病毒复制所必需的细胞功能提出了挑战。在这里，我们测量了宿主mRNA的翻译速率在牛痘病毒诱导的主机关闭通过分析总的和积极翻译mRNA在全基因组的方式。这项研究表明，氧化磷酸化的mRNA在牛痘病毒诱导的宿主蛋白质合成关闭过程中被上调。氧化磷酸化是主要的细胞能量产生途径，我们进一步表明，其功能的维护是重要的牛痘病毒复制。这项研究强调了这样一个事实，即牛痘病毒感染可以通过在整体宿主蛋白质合成关闭以满足能量消耗的背景下翻译上调来增强细胞能量产生。
ABSTRACT Vaccinia virus infection causes a host shutoff that is marked by global inhibition of host protein synthesis. Though the host shutoff may facilitate reallocation of cellular resources for viral replication and evasion of host antiviral immune responses, it poses a challenge for continuous synthesis of cellular proteins that are important for viral replication. It is, however, unclear whether and how certain cellular proteins may be selectively synthesized during the vaccinia virus-induced host shutoff. Using simultaneous RNA sequencing and ribosome profiling, two techniques quantifying genome-wide levels of mRNA and active protein translation, respectively, we analyzed the responses of host cells to vaccinia virus infection at both the transcriptional and translational levels. The analyses showed that cellular mRNA depletion played a dominant role in the shutoff of host protein synthesis. Though the cellular mRNAs were significantly reduced, the relative translation efficiency of a subset of cellular mRNAs increased, particularly those involved in oxidative phosphorylation that are responsible for cellular energy production. Further experiments demonstrated that the protein levels and activities of oxidative phosphorylation increased during vaccinia virus infection, while inhibition of the cellular oxidative phosphorylation function significantly suppressed vaccinia virus replication. Moreover, the short 5′ untranslated region of the oxidative phosphorylation mRNAs contributed to the translational upregulation. These results provide evidence of a mechanism that couples translational control and energy metabolism, two processes that all viruses depend on host cells to provide, to support vaccinia virus replication during a host shutoff. IMPORTANCE Many viral infections cause global host protein synthesis shutoff. While host protein synthesis shutoff benefits the virus by relocating cellular resources to viral replication, it also poses a challenge to the maintenance of cellular functions necessary for viral replication if continuous protein synthesis is required. Here we measured the host mRNA translation rate during a vaccinia virus-induced host shutoff by analyzing total and actively translating mRNAs in a genome-wide manner. This study revealed that oxidative phosphorylation mRNAs were translationally upregulated during vaccinia virus-induced host protein synthesis shutoff. Oxidative phosphorylation is the major cellular energy-producing pathway, and we further showed that maintenance of its function is important for vaccinia virus replication. This study highlights the fact that vaccinia virus infection can enhance cellular energy production through translational upregulation in the context of an overall host protein synthesis shutoff to meet energy expenditure.