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鉴定SARS-CoV-2基因组RNA的5'-近位转录和核囊蛋白之间的结合特征

Shih-Cheng Chen^1,2, Cui-Ting Xu², Chuan-Fu Chang²

¹National Institute of Cancer Research, National Health Research Institutes, Tainan City, Taiwan.

|March 13, 2025

概括

冠状病毒使用基因组RNA (gRNA) 上的包装信号 (PSs) 来通过核体 (N) 蛋白进行选择性包装. 这项研究揭示了SARS-CoV-2 5-UTR RNA和N蛋白之间的新结合特征,影响病毒RNA包装.

科学领域:

病毒学病毒学
分子生物学分子生物学
结构生物学结构生物学

背景情况:

冠状病毒 (CoV) 使用基因组RNA (gRNA) 上的包装信号 (PSs) 来通过核体 (N) 蛋白调节选择性包装.
虽然PSs在一些CoV中已知,但对它们的位置和相互作用机制的实验证据,特别是在SARS-CoV-2中,是有限的.

研究的目的:

为了研究严重急性呼吸系统综合征冠状病毒2 (SARS-CoV-2) 5'-近端gRNA转录和N蛋白之间的相互作用.
阐明参与N蛋白结合和病毒RNA包装的gRNA的结构特征.

主要方法:

电泳运动转移试验 (EMSAs) 用于研究RNA-蛋白相互作用.
在体外合成 CoV 5'-近端 gRNA 转录.
从SARS-CoV N蛋白质中识别RNA结合.
在体外观察N蛋白质凝聚物形成和液体-液体相分离 (LLPS).

主要成果:

SARS-CoV-2 5'-近端gRNA转录采用多种构造,具有富含茎环 (SL) 构造器,首选由N蛋白结合.
删除特定的5'-近位元素 (SL1,SL5a/b/c) 促进了替代RNA构造.
识别了与RNA结合的,优先结合野生型SL5aRNA.
有 CoV-5'-UTR 转录的 N 蛋白 LLPS 受 SL5a/b/c 的影响.

结论:

冠状病毒gRNAs的5'-近端区域表现出影响N蛋白结合的复杂结构动态.
特定的RNA结构元素,特别是SL5a/b/c,在N蛋白识别和潜在的液体-液体相分离中起着至关重要的作用.
这些发现揭示了对于理解冠状病毒RNA包装至关重要的新型结合特征.

关键词:

5-UTRs可以使用.N-蛋白质是一种N-蛋白质.结构 RNA 结构 RNA 结构具有 cis 作用的元素.冠状病毒冠状病毒的新冠病毒.封装封装封装封装它是核体的核体.

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