Herpes Simplex Virus 1 Lytic Infection Blocks MicroRNA (miRNA) Biogenesis at the Stage of Nuclear Export of Pre-miRNAs
Herpes Simplex Virus 1 Lytic Infection Blocks MicroRNA (miRNA) Biogenesis at the Stage of Nuclear Export of Pre-miRNAs
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单纯疱疹病毒 1 裂解感染在 Pre-miRNA 的核输出阶段阻断 MicroRNA (miRNA) 的生物发生
DOI：
10.1128/mbio.02856-18
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发表时间：
2019-02
期刊：
影响因子：
6.4
通讯作者：
Coen Donald M
中科院分区：
文献类型：
--
作者：
Pan Dongli;Li Gang;Morris Love Jenna;Qi Shuyuan;Feng Lei;Mertens Max E;Jurak Igor;Knipe David M;Coen Donald M
病毒针对宿主小RNA生物发生途径以达到最佳感染结果的各种机制已被确定。单纯疱疹病毒1 （HSV-1）是一种普遍存在的人类病原体，其在宿主体内的成功存在需要产生性（“溶解性”）和潜伏性感染。尽管已经发现了许多HSV-1 mirna，其中一些被认为有助于控制裂解/潜伏开关，但对其生物发生的调节知之甚少。通过对不同条件下pre-miRNA和成熟miRNA的表达进行表征，本研究揭示了裂解感染和潜伏感染之间miRNA生物发生的显著差异，并揭示了阻断pre-miRNA核输出的调控机制，该机制依赖于病毒蛋白ICP27和病毒DNA合成。这一机制代表了一种新的病毒-宿主相互作用，可以限制HSV-1 miRNA促进潜伏期的抑制作用，并可能揭示miRNA核输出的调控，这一机制一直相对未被探索。单纯疱疹病毒1 （HSV-1）在两种感染程序之间切换，生产（“溶解”）和潜伏感染。一些HSV-1 microrna （mirna）被假设有助于控制这种开关，但对其表达的调节知之甚少。通过Northern blot分析，我们发现，尽管六种HSV-1 miRNA的生物发生效率存在内在差异，但在不同细胞系的裂解感染过程中，以及在急性感染小鼠三叉神经节中检测到的情况下，所有六种miRNA/miRNA均表现出较高的前miRNA/miRNA比率。相比之下，在潜伏感染的神经节和用表达miRNA的慢病毒载体转导的细胞中观察到的比例要低得多，这表明HSV-1裂解性感染阻断了miRNA的生物发生。这种现象并不是病毒miRNA所特有的，因为重组HSV-1表达的宿主miRNA在裂解感染后期也表现出较高的pre-miRNA/miRNA比率。在放线菌素D存在的情况下，大多数成熟miRNA的水平在感染期间保持稳定，表明高比率是由于pre-miRNA转化为miRNA的效率低下。细胞分离实验表明，在感染后期（但不是早期），pre- mirna在细胞核中富集，在细胞质中减少，表明核输出被阻断。消除ICP27表达的突变或添加阿昔洛韦会降低pre-miRNA/miRNA比率，但突变会大幅降低Us11表达，但不会降低。因此，HSV-1裂解性感染在核输出阶段抑制miRNA的生物发生，并以ICP27和病毒DNA合成依赖的方式抑制miRNA的生物发生。这种机制可能通过在裂解感染期间降低抑制mirna的表达而使病毒受益，同时允许在潜伏期升高表达。病毒靶向宿主小RNA生物发生途径以达到最佳感染结果的各种机制已经被确定。单纯疱疹病毒1 （HSV-1）是一种普遍存在的人类病原体，其在宿主体内的成功存在需要产生性（“溶解性”）和潜伏性感染。尽管已经发现了许多HSV-1 mirna，其中一些被认为有助于控制裂解/潜伏开关，但对其生物发生的调节知之甚少。通过对不同条件下pre-miRNA和成熟miRNA的表达进行表征，本研究揭示了裂解感染和潜伏感染之间miRNA生物发生的显著差异，并揭示了阻断pre-miRNA核输出的调控机制，该机制依赖于病毒蛋白ICP27和病毒DNA合成。这一机制代表了一种新的病毒-宿主相互作用，可以限制HSV-1 miRNA促进潜伏期的抑制作用，并可能揭示miRNA核输出的调控，这一机制一直相对未被探索。
Various mechanisms have been identified by which viruses target host small RNA biogenesis pathways to achieve optimal infection outcomes. Herpes simplex virus 1 (HSV-1) is a ubiquitous human pathogen whose successful persistence in the host entails both productive (“lytic”) and latent infection. Although many HSV-1 miRNAs have been discovered and some are thought to help control the lytic/latent switch, little is known about regulation of their biogenesis. By characterizing expression of both pre-miRNAs and mature miRNAs under various conditions, this study revealed striking differences in miRNA biogenesis between lytic and latent infection and uncovered a regulatory mechanism that blocks pre-miRNA nuclear export and is dependent on viral protein ICP27 and viral DNA synthesis. This mechanism represents a new virus-host interaction that could limit the repressive effects of HSV-1 miRNAs hypothesized to promote latency and may shed light on the regulation of miRNA nuclear export, which has been relatively unexplored. ABSTRACT Herpes simplex virus 1 (HSV-1) switches between two infection programs, productive (“lytic”) and latent infection. Some HSV-1 microRNAs (miRNAs) have been hypothesized to help control this switch, and yet little is known about regulation of their expression. Using Northern blot analyses, we found that, despite inherent differences in biogenesis efficiency among six HSV-1 miRNAs, all six exhibited high pre-miRNA/miRNA ratios during lytic infection of different cell lines and, when detectable, in acutely infected mouse trigeminal ganglia. In contrast, considerably lower ratios were observed in latently infected ganglia and in cells transduced with lentiviral vectors expressing the miRNAs, suggesting that HSV-1 lytic infection blocks miRNA biogenesis. This phenomenon is not specific to viral miRNAs, as a host miRNA expressed from recombinant HSV-1 also exhibited high pre-miRNA/miRNA ratios late during lytic infection. The levels of most of the mature miRNAs remained stable during infection in the presence of actinomycin D, indicating that the high ratios are due to inefficient pre-miRNA conversion to miRNA. Cellular fractionation experiments showed that late (but not early) during infection, pre-miRNAs were enriched in the nucleus and depleted in the cytoplasm, indicating that nuclear export was blocked. A mutation eliminating ICP27 expression or addition of acyclovir reduced pre-miRNA/miRNA ratios, but mutations drastically reducing Us11 expression did not. Thus, HSV-1 lytic infection inhibits miRNA biogenesis at the step of nuclear export and does so in an ICP27- and viral DNA synthesis-dependent manner. This mechanism may benefit the virus by reducing expression of repressive miRNAs during lytic infection while permitting elevated expression during latency. IMPORTANCE Various mechanisms have been identified by which viruses target host small RNA biogenesis pathways to achieve optimal infection outcomes. Herpes simplex virus 1 (HSV-1) is a ubiquitous human pathogen whose successful persistence in the host entails both productive (“lytic”) and latent infection. Although many HSV-1 miRNAs have been discovered and some are thought to help control the lytic/latent switch, little is known about regulation of their biogenesis. By characterizing expression of both pre-miRNAs and mature miRNAs under various conditions, this study revealed striking differences in miRNA biogenesis between lytic and latent infection and uncovered a regulatory mechanism that blocks pre-miRNA nuclear export and is dependent on viral protein ICP27 and viral DNA synthesis. This mechanism represents a new virus-host interaction that could limit the repressive effects of HSV-1 miRNAs hypothesized to promote latency and may shed light on the regulation of miRNA nuclear export, which has been relatively unexplored.
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DOI：
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