Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation.
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 entir...
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Public Library of Science (PLoS)
2025-04-01
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| Series: | PLoS Biology |
| Online Access: | https://doi.org/10.1371/journal.pbio.3003115 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Hannah C Kubinski |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e1058cfde9b149899144790e960f36db |
| institution | OA Journals |
| issn | 1544-9173 1545-7885 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Biology |
| spelling | doaj-art-e1058cfde9b149899144790e960f36db2025-08-20T02:28:22ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852025-04-01234e300311510.1371/journal.pbio.3003115Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation.Hannah C KubinskiHannah W DespresBryan A JohnsonMadaline M SchmidtSara A JaffraniAllyson H TurnerConor D FanueleMargaret G MillsKumari G LokugamageCaroline M DumasDavid J ShirleyLeah K EstesAndrew PekoszJessica W CrothersPavitra RoychoudhuryAlexander L GreningerKeith R JeromeBruno Martorelli Di GenovaDavid H WalkerBryan A BallifMark S LadinskyPamela J BjorkmanVineet D MenacheryEmily A BruceThe 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.https://doi.org/10.1371/journal.pbio.3003115 |
| spellingShingle | 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 Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. PLoS Biology |
| title | Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. |
| title_full | Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. |
| title_fullStr | Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. |
| title_full_unstemmed | Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. |
| title_short | Variant mutation G215C in SARS-CoV-2 nucleocapsid enhances viral infection via altered genomic encapsidation. |
| title_sort | variant mutation g215c in sars cov 2 nucleocapsid enhances viral infection via altered genomic encapsidation |
| url | https://doi.org/10.1371/journal.pbio.3003115 |
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