Cargando…
Multiple Roles of SARS-CoV-2 N Protein Facilitated by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent Antibodies
[Image: see text] The SARS-CoV-2 nucleocapsid (N) protein is a highly immunogenic viral protein that plays essential roles in replication and virion assembly. Here, using native mass spectrometry, we show that dimers are the functional unit of ribonucleoprotein assembly and that N protein binds RNA...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American
Chemical Society
2021
|
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8231660/ https://www.ncbi.nlm.nih.gov/pubmed/34462738 http://dx.doi.org/10.1021/jacsau.1c00139 |
_version_ | 1783713476508647424 |
---|---|
author | Lutomski, Corinne A. El-Baba, Tarick J. Bolla, Jani R. Robinson, Carol V. |
author_facet | Lutomski, Corinne A. El-Baba, Tarick J. Bolla, Jani R. Robinson, Carol V. |
author_sort | Lutomski, Corinne A. |
collection | PubMed |
description | [Image: see text] The SARS-CoV-2 nucleocapsid (N) protein is a highly immunogenic viral protein that plays essential roles in replication and virion assembly. Here, using native mass spectrometry, we show that dimers are the functional unit of ribonucleoprotein assembly and that N protein binds RNA with a preference for GGG motifs, a common motif in coronavirus packaging signals. Unexpectedly, proteolytic processing of N protein resulted in the formation of additional proteoforms. The N-terminal proteoforms bind RNA, with the same preference for GGG motifs, and bind to cyclophilin A, an interaction which can be abolished by approved immunosuppressant cyclosporin A. Furthermore, N proteoforms showed significantly different interactions with IgM, IgG, and IgA antibodies from convalescent plasma. Notably, the C-terminal proteoform exhibited a heightened interaction with convalescent antibodies, suggesting the antigenic epitope is localized to the C-terminus. Overall, the different interactions of N proteoforms highlight potential avenues for therapeutic intervention and identify a stable and immunogenic proteoform as a possible candidate for immune-directed therapies. |
format | Online Article Text |
id | pubmed-8231660 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American
Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-82316602021-07-08 Multiple Roles of SARS-CoV-2 N Protein Facilitated by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent Antibodies Lutomski, Corinne A. El-Baba, Tarick J. Bolla, Jani R. Robinson, Carol V. JACS Au [Image: see text] The SARS-CoV-2 nucleocapsid (N) protein is a highly immunogenic viral protein that plays essential roles in replication and virion assembly. Here, using native mass spectrometry, we show that dimers are the functional unit of ribonucleoprotein assembly and that N protein binds RNA with a preference for GGG motifs, a common motif in coronavirus packaging signals. Unexpectedly, proteolytic processing of N protein resulted in the formation of additional proteoforms. The N-terminal proteoforms bind RNA, with the same preference for GGG motifs, and bind to cyclophilin A, an interaction which can be abolished by approved immunosuppressant cyclosporin A. Furthermore, N proteoforms showed significantly different interactions with IgM, IgG, and IgA antibodies from convalescent plasma. Notably, the C-terminal proteoform exhibited a heightened interaction with convalescent antibodies, suggesting the antigenic epitope is localized to the C-terminus. Overall, the different interactions of N proteoforms highlight potential avenues for therapeutic intervention and identify a stable and immunogenic proteoform as a possible candidate for immune-directed therapies. American Chemical Society 2021-06-15 /pmc/articles/PMC8231660/ /pubmed/34462738 http://dx.doi.org/10.1021/jacsau.1c00139 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Lutomski, Corinne A. El-Baba, Tarick J. Bolla, Jani R. Robinson, Carol V. Multiple Roles of SARS-CoV-2 N Protein Facilitated by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent Antibodies |
title | Multiple Roles of SARS-CoV-2 N Protein Facilitated
by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent
Antibodies |
title_full | Multiple Roles of SARS-CoV-2 N Protein Facilitated
by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent
Antibodies |
title_fullStr | Multiple Roles of SARS-CoV-2 N Protein Facilitated
by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent
Antibodies |
title_full_unstemmed | Multiple Roles of SARS-CoV-2 N Protein Facilitated
by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent
Antibodies |
title_short | Multiple Roles of SARS-CoV-2 N Protein Facilitated
by Proteoform-Specific Interactions with RNA, Host Proteins, and Convalescent
Antibodies |
title_sort | multiple roles of sars-cov-2 n protein facilitated
by proteoform-specific interactions with rna, host proteins, and convalescent
antibodies |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8231660/ https://www.ncbi.nlm.nih.gov/pubmed/34462738 http://dx.doi.org/10.1021/jacsau.1c00139 |
work_keys_str_mv | AT lutomskicorinnea multiplerolesofsarscov2nproteinfacilitatedbyproteoformspecificinteractionswithrnahostproteinsandconvalescentantibodies AT elbabatarickj multiplerolesofsarscov2nproteinfacilitatedbyproteoformspecificinteractionswithrnahostproteinsandconvalescentantibodies AT bollajanir multiplerolesofsarscov2nproteinfacilitatedbyproteoformspecificinteractionswithrnahostproteinsandconvalescentantibodies AT robinsoncarolv multiplerolesofsarscov2nproteinfacilitatedbyproteoformspecificinteractionswithrnahostproteinsandconvalescentantibodies |