Bermúdez-Méndez Erick, Katrukha Eugene A, Spruit Cindy M, Kortekaas Jeroen, Wichgers Schreur Paul J
Department of Virology, Wageningen Bioveterinary Research, Lelystad, The Netherlands.
Laboratory of Virology, Wageningen University, Wageningen, The Netherlands.
Commun Biol. 2021 Mar 22;4(1):345. doi: 10.1038/s42003-021-01821-y.
Bunyaviruses have a genome that is divided over multiple segments. Genome segmentation complicates the generation of progeny virus, since each newly formed virus particle should preferably contain a full set of genome segments in order to disseminate efficiently within and between hosts. Here, we combine immunofluorescence and fluorescence in situ hybridization techniques to simultaneously visualize bunyavirus progeny virions and their genomic content at single-molecule resolution in the context of singly infected cells. Using Rift Valley fever virus and Schmallenberg virus as prototype tri-segmented bunyaviruses, we show that bunyavirus genome packaging is influenced by the intracellular viral genome content of individual cells, which results in greatly variable packaging efficiencies within a cell population. We further show that bunyavirus genome packaging is more efficient in insect cells compared to mammalian cells and provide new insights on the possibility that incomplete particles may contribute to bunyavirus spread as well.
布尼亚病毒的基因组分布在多个片段上。基因组分段使子代病毒的产生变得复杂，因为每个新形成的病毒粒子最好包含一套完整的基因组片段，以便在宿主内部和宿主之间有效地传播。在这里，我们结合免疫荧光和荧光原位杂交技术，在单感染细胞的背景下，以单分子分辨率同时可视化布尼亚病毒子代病毒粒子及其基因组内容。使用裂谷热病毒和施马伦贝格病毒作为典型的三段式布尼亚病毒，我们表明布尼亚病毒基因组包装受单个细胞内病毒基因组含量的影响，这导致细胞群体内的包装效率差异很大。我们进一步表明，与哺乳动物细胞相比，布尼亚病毒基因组在昆虫细胞中的包装效率更高，并为不完整颗粒可能也有助于布尼亚病毒传播的可能性提供了新的见解。
