Time-Resolved Analysis of N-RNA Interactions during RVFV Infection Shows Qualitative and Quantitative Shifts in RNA Encapsidation and Packaging.

Time-Resolved Analysis of N-RNA Interactions during RVFV Infection Shows Qualitative and Quantitative Shifts in RNA Encapsidation and Packaging.
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RVFV感染过程中N-RNA相互作用的时间分辨分析表明,RNA的包膜和包装发生了质的和量的变化。

DOI:
10.3390/v13122417
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发表时间:
2021-12-02
期刊:
Viruses
影响因子:
--
通讯作者:
Lodmell JS
Lodmell JS
中科院分区:
其他
文献类型:
--
作者:
Hayashi M;Schultz EP;Lanchy JM;Lodmell JS

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裂谷热病毒 (RVFV) 是一种负义三联 RNA 病毒,在非洲和阿拉伯半岛流行。它可导致人类和家畜严重疾病和死亡,并且其在全球范围内传播的可能性令人担忧。 RVFV 的核衣壳蛋白 (N) 是一种 RNA 结合蛋白,是病毒转录、复制和新生病毒颗粒产生所必需的。我们进行了交联、免疫沉淀和测序 (CLIP-seq),以表征感染过程中 N 与宿主和病毒 RNA 的相互作用。同时,为了精确测量细胞内氮水平,我们采用了多反应监测质谱法 (MRM-MS)。我们的结果表明,N 在感染早期阶段主要与宿主 RNA 结合,产生感染性降低的新生病毒颗粒。 N 的表达在感染后 10 小时达到平台期,而细胞内病毒 RNA 浓度继续增加。而且,感染后期产生的病毒体具有更高的传染性。总而言之,对这些 N-RNA 相互作用的详细检查可以深入了解 N 和病毒 RNA 的调节表达如何产生感染性和不完整的非感染性颗粒。
Rift Valley fever virus (RVFV) is a negative-sense, tripartite RNA virus that is endemic to Africa and the Arabian Peninsula. It can cause severe disease and mortality in humans and domestic livestock and is a concern for its potential to spread more globally. RVFV’s nucleocapsid protein (N) is an RNA-binding protein that is necessary for viral transcription, replication, and the production of nascent viral particles. We have conducted crosslinking, immunoprecipitation, and sequencing (CLIP-seq) to characterize N interactions with host and viral RNAs during infection. In parallel, to precisely measure intracellular N levels, we employed multiple reaction monitoring mass spectrometry (MRM-MS). Our results show that N binds mostly to host RNAs at early stages of infection, yielding nascent virus particles of reduced infectivity. The expression of N plateaus 10 h post-infection, whereas the intracellular viral RNA concentration continues to increase. Moreover, the virions produced later in infection have higher infectivity. Taken together, the detailed examination of these N–RNA interactions provides insight into how the regulated expression of N and viral RNA produces both infectious and incomplete, noninfectious particles.
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