Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids
Summary: Nucleocapsid (N) protein of the SARS-CoV-2 virus packages the viral genome into well-defined ribonucleoprotein particles, but the molecular pathway is still unclear. N-protein is dimeric and consists of two folded domains with nucleic acid (NA) binding sites, surrounded by intrinsically dis...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004221004910 |
| _version_ | 1818441073985323008 |
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| author | Huaying Zhao Di Wu Ai Nguyen Yan Li Regina C. Adão Eugene Valkov George H. Patterson Grzegorz Piszczek Peter Schuck |
| author_facet | Huaying Zhao Di Wu Ai Nguyen Yan Li Regina C. Adão Eugene Valkov George H. Patterson Grzegorz Piszczek Peter Schuck |
| author_sort | Huaying Zhao |
| collection | DOAJ |
| description | Summary: Nucleocapsid (N) protein of the SARS-CoV-2 virus packages the viral genome into well-defined ribonucleoprotein particles, but the molecular pathway is still unclear. N-protein is dimeric and consists of two folded domains with nucleic acid (NA) binding sites, surrounded by intrinsically disordered regions that promote liquid-liquid phase separation. Here, we use biophysical tools to study N-protein interactions with oligonucleotides of different lengths, examining the size, composition, secondary structure, and energetics of the resulting states. We observe the formation of supramolecular clusters or nuclei preceding growth into phase-separated droplets. Short hexanucleotide NA forms compact 2:2 N-protein/NA complexes with reduced disorder. Longer oligonucleotides expose additional N-protein interactions and multi-valent protein-NA interactions, which generate higher-order mixed oligomers and simultaneously promote growth of droplets. Phase separation is accompanied by a significant change in protein secondary structure, different from that caused by initial NA binding, which may contribute to the assembly of ribonucleoprotein particles within macromolecular condensates. |
| first_indexed | 2024-12-14T18:22:28Z |
| format | Article |
| id | doaj.art-9763ec35145e4172ad30d879d8b02e32 |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-12-14T18:22:28Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-9763ec35145e4172ad30d879d8b02e322022-12-21T22:52:02ZengElsevieriScience2589-00422021-06-01246102523Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acidsHuaying Zhao0Di Wu1Ai Nguyen2Yan Li3Regina C. Adão4Eugene Valkov5George H. Patterson6Grzegorz Piszczek7Peter Schuck8Dynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, 13 South Drive, Bethesda, MD 20892, USABiophysics Core Facility, National Heart, Lung, and Blood Institute, 50 South Drive, Bethesda, MD 20892, USADynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, 13 South Drive, Bethesda, MD 20892, USAProtein/Peptide Sequencing Facility, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USADynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, 13 South Drive, Bethesda, MD 20892, USAMessenger RNA Regulation and Decay Section, RNA Biology Laboratory, Center for Cancer Research, National Cancer Institute, Building 560, Room 21-105A, Frederick, MD 21702, USASection on Biophotonics, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, MD 20892, USABiophysics Core Facility, National Heart, Lung, and Blood Institute, 50 South Drive, Bethesda, MD 20892, USADynamics of Macromolecular Assembly Section, Laboratory of Cellular Imaging and Macromolecular Biophysics, National Institute of Biomedical Imaging and Bioengineering, 13 South Drive, Bethesda, MD 20892, USA; Corresponding authorSummary: Nucleocapsid (N) protein of the SARS-CoV-2 virus packages the viral genome into well-defined ribonucleoprotein particles, but the molecular pathway is still unclear. N-protein is dimeric and consists of two folded domains with nucleic acid (NA) binding sites, surrounded by intrinsically disordered regions that promote liquid-liquid phase separation. Here, we use biophysical tools to study N-protein interactions with oligonucleotides of different lengths, examining the size, composition, secondary structure, and energetics of the resulting states. We observe the formation of supramolecular clusters or nuclei preceding growth into phase-separated droplets. Short hexanucleotide NA forms compact 2:2 N-protein/NA complexes with reduced disorder. Longer oligonucleotides expose additional N-protein interactions and multi-valent protein-NA interactions, which generate higher-order mixed oligomers and simultaneously promote growth of droplets. Phase separation is accompanied by a significant change in protein secondary structure, different from that caused by initial NA binding, which may contribute to the assembly of ribonucleoprotein particles within macromolecular condensates.http://www.sciencedirect.com/science/article/pii/S2589004221004910VirologyBiophysicsprotein folding |
| spellingShingle | Huaying Zhao Di Wu Ai Nguyen Yan Li Regina C. Adão Eugene Valkov George H. Patterson Grzegorz Piszczek Peter Schuck Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids iScience Virology Biophysics protein folding |
| title | Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids |
| title_full | Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids |
| title_fullStr | Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids |
| title_full_unstemmed | Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids |
| title_short | Energetic and structural features of SARS-CoV-2 N-protein co-assemblies with nucleic acids |
| title_sort | energetic and structural features of sars cov 2 n protein co assemblies with nucleic acids |
| topic | Virology Biophysics protein folding |
| url | http://www.sciencedirect.com/science/article/pii/S2589004221004910 |
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