Nucleocapsid mutations in SARS-CoV-2 augment replication and pathogenesis.
Nucleocapsid mutations in SARS-CoV-2 augment replication and pathogenesis.
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DOI：
10.1371/journal.ppat.1010627
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发表时间：
2022-06
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
作者：
虽然SARS-CoV-2继续适应人类感染和传播，但刺突基因以外的遗传变异在很大程度上仍未被探索。本研究调查了SARS-CoV-2核衣壳蛋白203-205位残基的高度可变区。在早期大流行（WA-1）背景下，重新创建在α和omicron变体中发现的突变，我们发现R203 K + G204 R突变足以增强SARS-CoV-2的复制，适应性和致病性。R203 K + G204 R突变体对应于体外和体内增加的病毒RNA和蛋白。重要的是，R203 K + G204 R突变增加了核衣壳磷酸化，并赋予对GSK-3激酶抑制的抗性，为增加病毒复制提供了分子基础。值得注意的是，在位置203+204处的类似丙氨酸取代也增加SARS-CoV-2复制并增强磷酸化，表明通过消除祖先的“RG”基序来增强感染。总的来说，这些结果表明，刺突外的变异突变是SARS-CoV-2持续适应人类感染的关键组成部分。自出现以来，SARS-CoV-2一直在适应人类感染，导致出现具有独特遗传特征的变体。大多数遗传变异的研究都集中在刺突上，这是目前可用疫苗的目标，而其他地方的变异的重要性研究不足。在这里，我们描述了一个高度可变的基序在残基203-205的核衣壳。在早期大流行背景下重现突出的核衣壳R203 K + G204 R突变，我们表明这种突变足以增强SARS-CoV-2的复制和致病性。我们还将R203 K + G204 R突变增强SARS-CoV-2感染与其调节核衣壳磷酸化联系起来。最后，我们描述了一个类似的丙氨酸双取代位置203-204。发现该突变体模拟R203 K + G204 R，表明通过破坏祖先序列发生感染增强。总之，我们的研究结果表明，穗以外的突变是SARS-CoV-2适应人类感染的关键组成部分。
While SARS-CoV-2 continues to adapt for human infection and transmission, genetic variation outside of the spike gene remains largely unexplored. This study investigates a highly variable region at residues 203–205 in the SARS-CoV-2 nucleocapsid protein. Recreating a mutation found in the alpha and omicron variants in an early pandemic (WA-1) background, we find that the R203K+G204R mutation is sufficient to enhance replication, fitness, and pathogenesis of SARS-CoV-2. The R203K+G204R mutant corresponds with increased viral RNA and protein both in vitro and in vivo. Importantly, the R203K+G204R mutation increases nucleocapsid phosphorylation and confers resistance to inhibition of the GSK-3 kinase, providing a molecular basis for increased virus replication. Notably, analogous alanine substitutions at positions 203+204 also increase SARS-CoV-2 replication and augment phosphorylation, suggesting that infection is enhanced through ablation of the ancestral ‘RG’ motif. Overall, these results demonstrate that variant mutations outside spike are key components in SARS-CoV-2’s continued adaptation to human infection. Since its emergence, SARS-CoV-2 has continued to adapt for human infection resulting in the emergence of variants with unique genetic profiles. Most studies of genetic variation have focused on spike, the target of currently available vaccines, leaving the importance of variation elsewhere understudied. Here, we characterize a highly variable motif at residues 203–205 in nucleocapsid. Recreating the prominent nucleocapsid R203K+G204R mutation in an early pandemic background, we show that this mutation is alone sufficient to enhance SARS-CoV-2 replication and pathogenesis. We also link augmentation of SARS-CoV-2 infection by the R203K+G204R mutation to its modulation of nucleocapsid phosphorylation. Finally, we characterize an analogous alanine double substitution at positions 203–204. This mutant was found to mimic R203K+G204R, suggesting augmentation of infection occurs by disrupting the ancestral sequence. Together, our findings illustrate that mutations outside of spike are key components of SARS-CoV-2’s adaptation to human infection.
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影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
64.5
作者：
Bouhaddou, Mehdi;Memon, Danish;Krogan, Nevan J.
通讯作者：
Krogan, Nevan J.
影响因子：
6.4
作者：
Josset L;Menachery VD;Gralinski LE;Agnihothram S;Sova P;Carter VS;Yount BL;Graham RL;Baric RS;Katze MG
通讯作者：
Katze MG
DOI：
10.3390/v6082991
发表时间：
2014-08-07
期刊：
Viruses
影响因子：
--
作者：
McBride R;van Zyl M;Fielding BC
通讯作者：
Fielding BC
影响因子：
11.8
作者：
Harcourt, Jennifer;Tamin, Azaibi;Thornburg, Natalie J.
通讯作者：
Thornburg, Natalie J.