@article{e2c1b27044c54842b844dab0858c6701,
title = "Nucleocapsid mutations in SARS-CoV-2 augment replication and pathogenesis",
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 {\textquoteleft}RG{\textquoteright} motif. Overall, these results demonstrate that variant mutations outside spike are key components in SARS-CoV-2{\textquoteright}s continued adaptation to human infection.",
author = "Johnson, {Bryan A.} and Yiyang Zhou and Lokugamage, {Kumari G.} and Vu, {Michelle N.} and Nathen Bopp and Crocquet-Valdes, {Patricia A.} and Birte Kalveram and Craig Schindewolf and Yang Liu and Dionna Scharton and Plante, {Jessica A.} and Xuping Xie and Patricia Aguilar and Weaver, {Scott C.} and Shi, {Pei Yong} and Walker, {David H.} and Routh, {Andrew L.} and Plante, {Kenneth S.} and Menachery, {Vineet D.}",
note = "Funding Information: Research was supported by grants from NIAID of the NIH (R01-AI153602 and R21-AI145400 to VDM; R24-AI120942 (WRCEVA) to SCW). ALR was supported by the Centers for Disease Control and Prevention (Contract 200-2021-11195). Both ALR and BAJ were supported by the Institute of Human Infection and Immunity at UTMB COVID-19 Research Fund. Research was also supported by STARs Award provided by the University of Texas System to VDM. Trainee funding provided by NIAID of the NIH to MNV (T32-AI060549) and to CS (T32-AI007526). BAJ was supported by the James W. McLaughlin Fellowship Fund. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: {\textcopyright} 2022 Johnson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",
year = "2022",
month = jun,
doi = "10.1371/journal.ppat.1010627",
language = "English (US)",
volume = "18",
journal = "PLoS Pathogens",
issn = "1553-7366",
publisher = "Public Library of Science",
number = "6",
}