tetano
Editor, Senior Moderator
Nat Commun
. 2025 Jul 15;16(1):6521.
doi: 10.1038/s41467-025-61861-4. Structural stabilization of the intrinsically disordered SARS-CoV-2 N by binding to RNA sequences engineered from the viral genome fragment
Sara Landeras-Bueno[SUP] 1 2 [/SUP], Chitra Hariharan[SUP] 3 [/SUP], Ruben Diaz Avalos[SUP] 3 [/SUP], Andrew S Norris[SUP] #[/SUP][SUP] 4 [/SUP], Dalton T Snyder[SUP] #[/SUP][SUP] 4 [/SUP], Kathryn M Hastie[SUP] 3 [/SUP], Stephanie Harkins[SUP] 3 [/SUP], Michelle Zandonatti[SUP] 3 [/SUP], Roshan R Rajamanickam[SUP] 3 [/SUP], Eduardo Olmedillas[SUP] 3 [/SUP], Robyn Miller[SUP] 3 [/SUP], Sujan Shresta[SUP] 3 [/SUP], Vicki H Wysocki[SUP] 4 [/SUP], Erica Ollmann Saphire[SUP] 5 6 [/SUP]
Affiliations
The nucleocapsid N is one of four structural proteins of the coronaviruses. Its essential role in genome encapsidation makes it a critical therapeutic target for COVID-19 and related diseases. However, the inherent disorder of full-length N hampers its structural analysis. Here, we describe a stepwise method using viral-derived RNAs to stabilize SARS-CoV-2 N for EM analysis. We identify pieces of RNA from the SARS-CoV-2 genome that promote the formation of structurally homogeneous N dimers, intermediates of assembly, and filamentous capsid-like structures. Building on these results, we engineer a symmetric RNA to stabilize N protein dimers, the building block of high-order assemblies, for EM studies. We combine domain-specific monoclonal antibodies against N with chemical cross-linking mass spectrometry to validate the spatial arrangement of the N domains within the dimer. Additionally, our cryo-EM analysis reveals novel antigenic sites on the N protein. Our findings provide insights into N protein´s architectural and antigenic principles, which can guide design of pan-coronavirus therapeutics.
. 2025 Jul 15;16(1):6521.
doi: 10.1038/s41467-025-61861-4. Structural stabilization of the intrinsically disordered SARS-CoV-2 N by binding to RNA sequences engineered from the viral genome fragment
Sara Landeras-Bueno[SUP] 1 2 [/SUP], Chitra Hariharan[SUP] 3 [/SUP], Ruben Diaz Avalos[SUP] 3 [/SUP], Andrew S Norris[SUP] #[/SUP][SUP] 4 [/SUP], Dalton T Snyder[SUP] #[/SUP][SUP] 4 [/SUP], Kathryn M Hastie[SUP] 3 [/SUP], Stephanie Harkins[SUP] 3 [/SUP], Michelle Zandonatti[SUP] 3 [/SUP], Roshan R Rajamanickam[SUP] 3 [/SUP], Eduardo Olmedillas[SUP] 3 [/SUP], Robyn Miller[SUP] 3 [/SUP], Sujan Shresta[SUP] 3 [/SUP], Vicki H Wysocki[SUP] 4 [/SUP], Erica Ollmann Saphire[SUP] 5 6 [/SUP]
Affiliations
- PMID: 40664703
- PMCID: PMC12264264
- DOI: 10.1038/s41467-025-61861-4
The nucleocapsid N is one of four structural proteins of the coronaviruses. Its essential role in genome encapsidation makes it a critical therapeutic target for COVID-19 and related diseases. However, the inherent disorder of full-length N hampers its structural analysis. Here, we describe a stepwise method using viral-derived RNAs to stabilize SARS-CoV-2 N for EM analysis. We identify pieces of RNA from the SARS-CoV-2 genome that promote the formation of structurally homogeneous N dimers, intermediates of assembly, and filamentous capsid-like structures. Building on these results, we engineer a symmetric RNA to stabilize N protein dimers, the building block of high-order assemblies, for EM studies. We combine domain-specific monoclonal antibodies against N with chemical cross-linking mass spectrometry to validate the spatial arrangement of the N domains within the dimer. Additionally, our cryo-EM analysis reveals novel antigenic sites on the N protein. Our findings provide insights into N protein´s architectural and antigenic principles, which can guide design of pan-coronavirus therapeutics.