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Assembly of SARS-CoV-2 nucleocapsid protein with nucleic acid.
- Source :
-
Nucleic acids research [Nucleic Acids Res] 2024 Jun 24; Vol. 52 (11), pp. 6647-6661. - Publication Year :
- 2024
-
Abstract
- The viral genome of SARS-CoV-2 is packaged by the nucleocapsid (N-)protein into ribonucleoprotein particles (RNPs), 38 ± 10 of which are contained in each virion. Their architecture has remained unclear due to the pleomorphism of RNPs, the high flexibility of N-protein intrinsically disordered regions, and highly multivalent interactions between viral RNA and N-protein binding sites in both N-terminal (NTD) and C-terminal domain (CTD). Here we explore critical interaction motifs of RNPs by applying a combination of biophysical techniques to ancestral and mutant proteins binding different nucleic acids in an in vitro assay for RNP formation, and by examining nucleocapsid protein variants in a viral assembly assay. We find that nucleic acid-bound N-protein dimers oligomerize via a recently described protein-protein interface presented by a transient helix in its long disordered linker region between NTD and CTD. The resulting hexameric complexes are stabilized by multivalent protein-nucleic acid interactions that establish crosslinks between dimeric subunits. Assemblies are stabilized by the dimeric CTD of N-protein offering more than one binding site for stem-loop RNA. Our study suggests a model for RNP assembly where N-protein scaffolding at high density on viral RNA is followed by cooperative multimerization through protein-protein interactions in the disordered linker.<br /> (Published by Oxford University Press on behalf of Nucleic Acids Research 2024.)
- Subjects :
- Protein Binding
Binding Sites
Ribonucleoproteins metabolism
Ribonucleoproteins chemistry
Ribonucleoproteins genetics
Virus Assembly genetics
Humans
Nucleocapsid Proteins chemistry
Nucleocapsid Proteins metabolism
Nucleocapsid Proteins genetics
Models, Molecular
Phosphoproteins chemistry
Phosphoproteins metabolism
Phosphoproteins genetics
COVID-19 virology
SARS-CoV-2 genetics
SARS-CoV-2 metabolism
SARS-CoV-2 chemistry
Coronavirus Nucleocapsid Proteins chemistry
Coronavirus Nucleocapsid Proteins metabolism
Coronavirus Nucleocapsid Proteins genetics
RNA, Viral metabolism
RNA, Viral chemistry
RNA, Viral genetics
Protein Multimerization
Subjects
Details
- Language :
- English
- ISSN :
- 1362-4962
- Volume :
- 52
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Nucleic acids research
- Publication Type :
- Academic Journal
- Accession number :
- 38587193
- Full Text :
- https://doi.org/10.1093/nar/gkae256