Dissecting Herpes Simplex Virus 1-Induced Host Shutoff at the RNA Level.
Dissecting Herpes Simplex Virus 1-Induced Host Shutoff at the RNA Level.
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DOI：
10.1128/jvi.01399-20
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发表时间：
2021-01-13
影响因子：
5.4
通讯作者：
Dölken L
中科院分区：
文献类型：
--
作者：
Friedel CC;Whisnant AW;Djakovic L;Rutkowski AJ;Friedl MS;Kluge M;Williamson JC;Sai S;Vidal RO;Sauer S;Hennig T;Grothey A;Milić A;Prusty BK;Lehner PJ;Matheson NJ;Erhard F;Dölken L
HSV-1病毒粒子宿主关闭(VHS)蛋白以翻译依赖的方式有效地切割宿主和病毒的mRNAs。在这项研究中，我们模拟和量化了VHS活性的变化，以及病毒诱导的宿主转录活性的全局丧失，在生产性HSV-1感染期间。总体而言，HSV-1诱导的总RNA水平的变化主要受这两种全局效应的影响。相比之下，染色质相关的RNA描述了基因特有的转录变化。这揭示了在WT和ΔVHS感染中高度一致的转录变化，证实了DUX4是HSV1感染中的关键转录调节因子，并发现VHS依赖的整合素黏附和细胞外基质成分的转录下调。后者解释了以前似乎归因于VHS介导的mRNA降解的基因特异性效应，并导致在8h P.I.时蛋白质水平的一致性丢失。许多各自的基因。单纯疱疹病毒1型(HSV-1)在裂解感染过程中会引起宿主的严重关闭。病毒粒子宿主关闭蛋白(VHS)通过有效地切割宿主和病毒的mRNAs而在这一过程中发挥关键作用。此外，病毒DNA复制的开始伴随着宿主转录活性的迅速下降。为了分析这两种机制的相对作用，并阐明在单纯疱疹病毒1型裂解感染的前8h内宿主的基因特异性转录反应，我们使用了野生型(WT)和Δvhs突变株感染原代人成纤维细胞的总的、新转录的(4SU标记的)和染色质相关的RNA的转录组测序。病毒进入后，VHS活性以每小时约30%的细胞mRNAs清除速率迅速稳定，直到感染后8小时(P.I.)。同时，宿主转录活性下降到10%到20%。虽然这两种现象的综合作用主导了感染诱导的总核糖核酸的变化，但在染色质相关核糖核酸中可以观察到广泛的基因特异性转录调控，并且在WT和ΔVHS感染之间令人惊讶地一致。两者都诱导了一小部分基因的强烈转录上调，这些基因在感染前表达很低，但已经被其启动子上的H3K4me3组蛋白标记启动。最有趣的是，对染色质相关RNA的分析显示，VHS核酸酶活性依赖于至少150个细胞基因的转录下调，特别是许多整合素黏附和细胞外基质成分。这伴随着依赖VHS的蛋白质水平在8h P.I.下降。这些基因中的许多。综上所述，我们的研究提供了在HSV-1裂解感染的前8小时内控制细胞RNA代谢的分子机制的全面图景。重要性HSV-1病毒粒子宿主关闭(VHS)蛋白以翻译依赖的方式有效地切割宿主和病毒的mRNAs。在这项研究中，我们模拟和量化了VHS活性的变化，以及病毒诱导的宿主转录活性的全局丧失，在生产性HSV-1感染期间。总体而言，HSV-1诱导的总RNA水平的变化主要受这两种全局效应的影响。相比之下，染色质相关的RNA描述了基因特有的转录变化。这揭示了在WT和ΔVHS感染中高度一致的转录变化，证实了DUX4是HSV1感染中的关键转录调节因子，并发现VHS依赖的整合素黏附和细胞外基质成分的转录下调。后者解释了以前似乎归因于VHS介导的mRNA降解的基因特异性效应，并导致在8h P.I.时蛋白质水平的一致性丢失。许多各自的基因。
The HSV-1 virion host shutoff (vhs) protein efficiently cleaves both host and viral mRNAs in a translation-dependent manner. In this study, we model and quantify changes in vhs activity, as well as virus-induced global loss of host transcriptional activity, during productive HSV-1 infection. In general, HSV-1-induced alterations in total RNA levels were dominated by these two global effects. In contrast, chromatin-associated RNA depicted gene-specific transcriptional changes. This revealed highly concordant transcriptional changes in WT and Δvhs infections, confirmed DUX4 as a key transcriptional regulator in HSV-1 infection, and identified vhs-dependent transcriptional downregulation of the integrin adhesome and extracellular matrix components. The latter explained seemingly gene-specific effects previously attributed to vhs-mediated mRNA degradation and resulted in a concordant loss in protein levels by 8 h p.i. for many of the respective genes. Herpes simplex virus 1 (HSV-1) induces a profound host shutoff during lytic infection. The virion host shutoff (vhs) protein plays a key role in this process by efficiently cleaving host and viral mRNAs. Furthermore, the onset of viral DNA replication is accompanied by a rapid decline in host transcriptional activity. To dissect relative contributions of both mechanisms and elucidate gene-specific host transcriptional responses throughout the first 8 h of lytic HSV-1 infection, we used transcriptome sequencing of total, newly transcribed (4sU-labeled) and chromatin-associated RNA in wild-type (WT) and Δvhs mutant infection of primary human fibroblasts. Following virus entry, vhs activity rapidly plateaued at an elimination rate of around 30% of cellular mRNAs per hour until 8 h postinfection (p.i.). In parallel, host transcriptional activity dropped to 10 to 20%. While the combined effects of both phenomena dominated infection-induced changes in total RNA, extensive gene-specific transcriptional regulation was observable in chromatin-associated RNA and was surprisingly concordant between WT and Δvhs infections. Both induced strong transcriptional upregulation of a small subset of genes that were poorly expressed prior to infection but already primed by H3K4me3 histone marks at their promoters. Most interestingly, analysis of chromatin-associated RNA revealed vhs-nuclease-activity-dependent transcriptional downregulation of at least 150 cellular genes, in particular of many integrin adhesome and extracellular matrix components. This was accompanied by a vhs-dependent reduction in protein levels by 8 h p.i. for many of these genes. In summary, our study provides a comprehensive picture of the molecular mechanisms that govern cellular RNA metabolism during the first 8 h of lytic HSV-1 infection. IMPORTANCE The HSV-1 virion host shutoff (vhs) protein efficiently cleaves both host and viral mRNAs in a translation-dependent manner. In this study, we model and quantify changes in vhs activity, as well as virus-induced global loss of host transcriptional activity, during productive HSV-1 infection. In general, HSV-1-induced alterations in total RNA levels were dominated by these two global effects. In contrast, chromatin-associated RNA depicted gene-specific transcriptional changes. This revealed highly concordant transcriptional changes in WT and Δvhs infections, confirmed DUX4 as a key transcriptional regulator in HSV-1 infection, and identified vhs-dependent transcriptional downregulation of the integrin adhesome and extracellular matrix components. The latter explained seemingly gene-specific effects previously attributed to vhs-mediated mRNA degradation and resulted in a concordant loss in protein levels by 8 h p.i. for many of the respective genes.