RT Journal Article
SR Electronic
T1 Structural basis of RNA recognition by the SARS-CoV-2 nucleocapsid phosphoprotein
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.04.02.022194
DO 10.1101/2020.04.02.022194
A1 Dhurvas Chandrasekaran Dinesh
A1 Dominika Chalupska
A1 Jan Silhan
A1 Vaclav Veverka
A1 Evzen Boura
YR 2020
UL http://biorxiv.org/content/early/2020/04/05/2020.04.02.022194.abstract
AB Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the Coronavirus disease 2019 (COVID-19) which is currently negatively affecting the population and disrupting the global economy. SARS-CoV-2 belongs to the +RNA virus family that utilize single-stranded positive-sense RNA molecules as genomes. SARS-CoV-2, like other coronaviruses, has an unusually large genome for a +RNA virus that encodes four structural proteins – the matrix (M), small envelope (E), spike (S) and nucleocapsid phosphoprotein (N) - and sixteen nonstructural proteins (nsp1-16) that together ensure replication of the virus in the host cell. The nucleocapsid phosphoprotein N is essential for linking the viral genome to the viral membrane. Its N-terminal RNA binding domain (N-NTD) captures the RNA genome while the C-terminal domain anchors the ribonucleoprotein complex to the viral membrane via its interaction with the M protein. Here, we characterized the structure of the N-NTD and its interaction with RNA using NMR spectroscopy. We observed a positively charged canyon on the surface of the N-NTD lined with arginine residues suggesting a putative RNA binding site. Next, we performed an NMR titration experiment using an RNA duplex. The observed changes in positions of signals in the N-NTD NMR spectra allowed us to construct a model of the N-NTD in complex with RNA.