1. Nucleocapsid mutations in SARS-CoV-2 augment replication and pathogenesis.
- Author
-
Johnson BA, Zhou Y, Lokugamage KG, Vu MN, Bopp N, Crocquet-Valdes PA, Kalveram B, Schindewolf C, Liu Y, Scharton D, Plante JA, Xie X, Aguilar P, Weaver SC, Shi PY, Walker DH, Routh AL, Plante KS, and Menachery VD
- Subjects
- Glycogen Synthase Kinase 3, Humans, Mutation, Nucleocapsid, Spike Glycoprotein, Coronavirus genetics, COVID-19 genetics, SARS-CoV-2 genetics
- Abstract
While SARS-CoV-2 continues to adapt for human infection and transmission, genetic variation outside of the spike gene remains largely unexplored. This study investigates a highly variable region at residues 203-205 in the SARS-CoV-2 nucleocapsid protein. Recreating a mutation found in the alpha and omicron variants in an early pandemic (WA-1) background, we find that the R203K+G204R mutation is sufficient to enhance replication, fitness, and pathogenesis of SARS-CoV-2. The R203K+G204R mutant corresponds with increased viral RNA and protein both in vitro and in vivo. Importantly, the R203K+G204R mutation increases nucleocapsid phosphorylation and confers resistance to inhibition of the GSK-3 kinase, providing a molecular basis for increased virus replication. Notably, analogous alanine substitutions at positions 203+204 also increase SARS-CoV-2 replication and augment phosphorylation, suggesting that infection is enhanced through ablation of the ancestral 'RG' motif. Overall, these results demonstrate that variant mutations outside spike are key components in SARS-CoV-2's continued adaptation to human infection., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: X.X., P-Y. S, and V.D.M. have filed a patent on the reverse genetic system and reporter SARS-CoV-2. Other authors declare no competing interests.
- Published
- 2022
- Full Text
- View/download PDF