TY - JOUR
T1 - Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain
AU - Wu, Chao
AU - Qavi, Abraham J.
AU - Hachim, Asmaa
AU - Kavian, Niloufar
AU - Cole, Aidan R.
AU - Moyle, Austin B.
AU - Wagner, Nicole D.
AU - Sweeney-Gibbons, Joyce
AU - Rohrs, Henry W.
AU - Gross, Michael L.
AU - Peiris, J. S.Malik
AU - Basler, Christopher F.
AU - Farnsworth, Christopher W.
AU - Valkenburg, Sophie A.
AU - Amarasinghe, Gaya K.
AU - Leung, Daisy W.
N1 - Funding Information:
We thank Dr. N. Krogan (UCSF) for sharing SARS-CoV-2 plasmids, Drs. A. Holehouse and A. Soranno (Washington University School of Medicine) for providing critical feedback, and R. Ridings for coordinating studies between WUSM and external groups. We would like to thank Bruker for mass spectrometry technical and instrument support, and Protein Metrics (R42GM1213302 to H.W.R) and Sierra Analytics for providing proteomic and HDX data analysis software. Research was supported by Fast Grant #2161 (Emergent Ventures) to G.K.A. Fast Grant # 2158 to C.F.B, and NIH grants (P01AI120943, R01AI123926 to G.K.A. and C.F.B; R01AI107056 to D.W.L.; P41GM103422 and R24GM136766 to M.L.G.; R01AI143292 and R01AI148663 C.F.B). S.A.V was supported by COVID190115 and COVID190126 Health and Medical Research Fund, Food and Health Bureau, Hong Kong and NIH/NIAID CEIRS contract HHSN272201400006C. A.J.Q. is supported by a NIH T32 training grant (T32CA009547). CW, GKA, and DWL conceived the overall project. All authors were integral to the design and execution of the study. CW, AQ, GKA, and DWL wrote the initial draft with significant input from all authors. The authors declare no competing interests. We worked to ensure sex balance in the selection of non-human subjects. One or more of the authors of this paper self-identifies as living with a disability. One or more of the authors of this paper received support from a program designed to increase minority representation in science.
Funding Information:
We thank Dr. N. Krogan (UCSF) for sharing SARS-CoV-2 plasmids, Drs. A. Holehouse and A. Soranno (Washington University School of Medicine) for providing critical feedback, and R. Ridings for coordinating studies between WUSM and external groups. We would like to thank Bruker for mass spectrometry technical and instrument support, and Protein Metrics (R42GM1213302 to H.W.R) and Sierra Analytics for providing proteomic and HDX data analysis software. Research was supported by Fast Grant #2161 (Emergent Ventures) to G.K.A., Fast Grant # 2158 to C.F.B, and NIH grants ( P01AI120943 , R01AI123926 to G.K.A. and C.F.B; R01AI107056 to D.W.L.; P41GM103422 and R24GM136766 to M.L.G.; R01AI143292 and R01AI148663 C.F.B). S.A.V was supported by COVID190115 and COVID190126 Health and Medical Research Fund, Food and Health Bureau, Hong Kong and NIH /NIAID CEIRS contract HHSN272201400006C. A.J.Q. is supported by a NIH T32 training grant ( T32CA009547 ).
Publisher Copyright:
© 2021 The Author(s)
PY - 2021/6/25
Y1 - 2021/6/25
N2 - Nucleocapsid (N) encoded by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays key roles in the replication cycle and is a critical serological marker. Here, we characterize essential biochemical properties of N and describe the utility of these insights in serological studies. We define N domains important for oligomerization and RNA binding and show that N oligomerization provides a high-affinity RNA-binding platform. We also map the RNA-binding interface, showing protection in the N-terminal domain and linker region. In addition, phosphorylation causes reduction of RNA binding and redistribution of N from liquid droplets to loose coils, showing how N-RNA accessibility and assembly may be regulated by phosphorylation. Finally, we find that the C-terminal domain of N is the most immunogenic, based on antibody binding to patient samples. Together, we provide a biochemical description of SARS-CoV-2 N and highlight the value of using N domains as highly specific and sensitive diagnostic markers.
AB - Nucleocapsid (N) encoded by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays key roles in the replication cycle and is a critical serological marker. Here, we characterize essential biochemical properties of N and describe the utility of these insights in serological studies. We define N domains important for oligomerization and RNA binding and show that N oligomerization provides a high-affinity RNA-binding platform. We also map the RNA-binding interface, showing protection in the N-terminal domain and linker region. In addition, phosphorylation causes reduction of RNA binding and redistribution of N from liquid droplets to loose coils, showing how N-RNA accessibility and assembly may be regulated by phosphorylation. Finally, we find that the C-terminal domain of N is the most immunogenic, based on antibody binding to patient samples. Together, we provide a biochemical description of SARS-CoV-2 N and highlight the value of using N domains as highly specific and sensitive diagnostic markers.
KW - Biophysics
KW - Human Specimen
KW - Virology
UR - http://www.scopus.com/inward/record.url?scp=85108340672&partnerID=8YFLogxK
U2 - 10.1016/j.isci.2021.102681
DO - 10.1016/j.isci.2021.102681
M3 - Article
C2 - 34095780
AN - SCOPUS:85108340672
SN - 2589-0042
VL - 24
JO - iScience
JF - iScience
IS - 6
M1 - 102681
ER -