Defective Viral Genomes Alter How Sendai Virus Interacts with Cellular Trafficking Machinery, Leading to Heterogeneity in the Production of Viral Particles among Infected Cells
Defective Viral Genomes Alter How Sendai Virus Interacts with Cellular Trafficking Machinery, Leading to Heterogeneity in the Production of Viral Particles among Infected Cells
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DOI：
10.1128/jvi.01579-18
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发表时间：
2019-02-01
影响因子：
5.4
通讯作者：
Lopez, Carolina B.
中科院分区：
文献类型：
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作者：
Genoyer, Emmanuelle;Lopez, Carolina B.
在RNA病毒复制过程中产生的缺陷病毒基因组（DVG）通过触发先天免疫、降低毒力以及在许多情况下促进持续感染的建立来决定感染结果。尽管它们在病毒与宿主相互作用中发挥着关键作用，但人们对调节DVGs产生和繁殖的机制知之甚少。可视化的病毒基因组使用RNA荧光原位杂交显示了显着的差异，在细胞内定位的DVG和全长病毒基因组感染副粘病毒仙台病毒。在富含全长病毒的细胞中，病毒基因组聚集在核周区域，并与细胞运输机制相关，包括微管和GTCRab 11 a。然而，在富含DVG的细胞中，有缺陷的基因组弥漫地分布在整个细胞质中，并且未能与这种细胞机制相互作用。因此，富含全长基因组的细胞产生含有DVG和全长基因组的病毒颗粒，而DVG高细胞产生的病毒颗粒很差，但强烈刺激抗病毒免疫。这些发现揭示了标准和含DVG的颗粒的选择性生产由受感染细胞的亚群，其可以通过DVG的细胞内定位来区分。这项研究强调了在分析感染过程中病毒-宿主相互作用时考虑这种功能异质性的重要性。重要意义仙台病毒感染过程中产生的缺陷病毒基因组（DVG）在一些感染细胞的细胞质中积累，并刺激抗病毒免疫和细胞存活。DVG作为缺陷颗粒进行包装和放行，对感染结局有显著影响。我们表明，DVG高细胞的亚群参与病毒包装和出芽机制较差，不能有效地产生病毒颗粒。相反，在感染过程中，富含全长基因组的细胞是标准和缺陷病毒颗粒的主要生产者。这项研究证明了感染细胞内发生的分子相互作用的异质性，并强调了细胞作为免疫启动子或病毒颗粒的生产者和永存者的不同功能作用，这取决于它们的病毒基因组内容及其细胞内定位。
Defective viral genomes (DVGs) generated during RNA virus replication determine infection outcome by triggering innate immunity, diminishing virulence, and, in many cases, facilitating the establishment of persistent infections. Despite their critical role during virus-host interactions, the mechanisms regulating the production and propagation of DVGs are poorly understood. Visualization of viral genomes using RNA fluorescent in situ hybridization revealed a striking difference in the intracellular localization of DVGs and full-length viral genomes during infections with the paramyxovirus Sendai virus. In cells enriched in full-length virus, viral genomes clustered in a perinuclear region and associated with cellular trafficking machinery, including microtubules and the GTPase Rab11a. However, in cells enriched in DVGs, defective genomes distributed diffusely throughout the cytoplasm and failed to interact with this cellular machinery. Consequently, cells enriched in full-length genomes produced both DVG- and full-length-genome-containing viral particles, while DVG-high cells poorly produced viral particles yet strongly stimulated antiviral immunity. These findings reveal the selective production of both standard and DVG-containing particles by a subpopulation of infected cells that can be differentiated by the intracellular localization of DVGs. This study highlights the importance of considering this functional heterogeneity in analyses of virus-host interactions during infection.IMPORTANCE Defective viral genomes (DVGs) generated during Sendai virus infections accumulate in the cytoplasm of some infected cells and stimulate antiviral immunity and cell survival. DVGs are packaged and released as defective particles and have a significant impact on infection outcome. We show that the subpopulation of DVG-high cells poorly engages the virus packaging and budding machinery and do not effectively produce viral particles. In contrast, cells enriched in full-length genomes are the primary producers of both standard and defective viral particles during infection. This study demonstrates heterogeneity in the molecular interactions occurring within infected cells and highlights distinct functional roles for cells as either initiators of immunity or producers and perpetuators of viral particles depending on their content of viral genomes and their intracellular localization.