Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation.

IF 9.8 1区 生物学 Q1 Agricultural and Biological Sciences
PLoS Biology Pub Date : 2025-04-29 eCollection Date: 2025-04-01 DOI:10.1371/journal.pbio.3003115
Hannah C Kubinski, Hannah W Despres, Bryan A Johnson, Madaline M Schmidt, Sara A Jaffrani, Allyson H Turner, Conor D Fanuele, Margaret G Mills, Kumari G Lokugamage, Caroline M Dumas, David J Shirley, Leah K Estes, Andrew Pekosz, Jessica W Crothers, Pavitra Roychoudhury, Alexander L Greninger, Keith R Jerome, Bruno Martorelli Di Genova, David H Walker, Bryan A Ballif, Mark S Ladinsky, Pamela J Bjorkman, Vineet D Menachery, Emily A Bruce
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引用次数: 0

Abstract

The evolution of SARS-CoV-2 variants and their respective phenotypes represents an important set of tools to understand basic coronavirus biology as well as the public health implications of individual mutations in variants of concern. While mutations outside of spike are not well studied, the entire viral genome is undergoing evolutionary selection, with several variants containing mutations in the central disordered linker region of the nucleocapsid (N) protein. Here, we identify a mutation (G215C), characteristic of the Delta variant, that introduces a novel cysteine into this linker domain, which results in the formation of a more stable N-N dimer. Using reverse genetics, we determined that this cysteine residue is necessary and sufficient for stable dimer formation in a WA1 SARS-CoV-2 background, where it results in significantly increased viral growth both in vitro and in vivo. Mechanistically, we show that the N:G215C mutant has more encapsidation as measured by increased RNA binding to N, N incorporation into virions, and electron microscopy showing that individual virions are larger, with elongated morphologies.

SARS-CoV-2核衣壳变异突变G215C通过改变基因组衣壳增强病毒感染
SARS-CoV-2变异体的进化及其各自的表型是一套重要的工具,可以了解基本的冠状病毒生物学以及受关注的变异体中个体突变的公共卫生影响。虽然刺突外的突变还没有得到很好的研究,但整个病毒基因组正在经历进化选择,在核衣壳(N)蛋白的中心无序连接区有几种变体含有突变。在这里,我们发现了一个突变(G215C),这是Delta变体的特征,它将一种新的半胱氨酸引入该连接体结构域,从而形成更稳定的N-N二聚体。利用反向遗传学,我们确定在WA1 SARS-CoV-2背景下,这种半胱氨酸残基对于稳定的二聚体形成是必要的和充分的,它导致体外和体内的病毒生长显著增加。从机制上讲,我们发现N:G215C突变体具有更多的囊化,通过增加RNA与N的结合,N并入病毒粒子,以及电子显微镜显示单个病毒粒子更大,具有细长的形态来测量。
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来源期刊
PLoS Biology
PLoS Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-BIOLOGY
CiteScore
15.40
自引率
2.00%
发文量
359
审稿时长
3-8 weeks
期刊介绍: PLOS Biology is the flagship journal of the Public Library of Science (PLOS) and focuses on publishing groundbreaking and relevant research in all areas of biological science. The journal features works at various scales, ranging from molecules to ecosystems, and also encourages interdisciplinary studies. PLOS Biology publishes articles that demonstrate exceptional significance, originality, and relevance, with a high standard of scientific rigor in methodology, reporting, and conclusions. The journal aims to advance science and serve the research community by transforming research communication to align with the research process. It offers evolving article types and policies that empower authors to share the complete story behind their scientific findings with a diverse global audience of researchers, educators, policymakers, patient advocacy groups, and the general public. PLOS Biology, along with other PLOS journals, is widely indexed by major services such as Crossref, Dimensions, DOAJ, Google Scholar, PubMed, PubMed Central, Scopus, and Web of Science. Additionally, PLOS Biology is indexed by various other services including AGRICOLA, Biological Abstracts, BIOSYS Previews, CABI CAB Abstracts, CABI Global Health, CAPES, CAS, CNKI, Embase, Journal Guide, MEDLINE, and Zoological Record, ensuring that the research content is easily accessible and discoverable by a wide range of audiences.
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