Genomic RNA Elements Drive Phase Separation of the SARS-CoV-2 Nucleocapsid.

TitleGenomic RNA Elements Drive Phase Separation of the SARS-CoV-2 Nucleocapsid.
Publication TypeJournal Article
Year of Publication2020
AuthorsIserman, C, Roden, CA, Boerneke, MA, Sealfon, RSG, McLaughlin, GA, Jungreis, I, Fritch, EJ, Hou, YJ, Ekena, J, Weidmann, CA, Theesfeld, CL, Kellis, M, Troyanskaya, OG, Baric, RS, Sheahan, TP, Weeks, KM, Gladfelter, AS
JournalMol Cell
Volume80
Issue6
Pagination1078-1091.e6
Date Published2020 12 17
ISSN1097-4164
KeywordsAnimals, Antiviral Agents, Chlorocebus aethiops, Coronavirus Nucleocapsid Proteins, COVID-19, Drug Evaluation, Preclinical, Genome, Viral, HEK293 Cells, Humans, Nucleocapsid, Phosphoproteins, RNA, Viral, SARS-CoV-2, Vero Cells
Abstract

<p>We report that the SARS-CoV-2 nucleocapsid protein (N-protein) undergoes liquid-liquid phase separation (LLPS) with viral RNA. N-protein condenses with specific RNA genomic elements under physiological buffer conditions and condensation is enhanced at human body temperatures (33°C and 37°C) and reduced at room temperature (22°C). RNA sequence and structure in specific genomic regions regulate N-protein condensation while other genomic regions promote condensate dissolution, potentially preventing aggregation of the large genome. At low concentrations, N-protein preferentially crosslinks to specific regions characterized by single-stranded RNA flanked by structured elements and these features specify the location, number, and strength of N-protein binding sites (valency). Liquid-like N-protein condensates form in mammalian cells in a concentration-dependent manner and can be altered by small molecules. Condensation of N-protein is RNA sequence and structure specific, sensitive to human body temperature, and manipulatable with small molecules, and therefore presents a screenable process for identifying antiviral compounds effective against SARS-CoV-2.</p>

DOI10.1016/j.molcel.2020.11.041
Alternate JournalMol Cell
PubMed ID33290746
PubMed Central IDPMC7691212
Grant ListF32 GM128330 / GM / NIGMS NIH HHS / United States
T32 CA009156 / CA / NCI NIH HHS / United States
U54 HL117798 / HL / NHLBI NIH HHS / United States
R01 HL111527 / HL / NHLBI NIH HHS / United States
R35 GM122532 / GM / NIGMS NIH HHS / United States
R01 GM071966 / GM / NIGMS NIH HHS / United States
U19 AI142759 / AI / NIAID NIH HHS / United States
R01 GM081506 / GM / NIGMS NIH HHS / United States
R01 HG005998 / HG / NHGRI NIH HHS / United States
F32 GM136164 / GM / NIGMS NIH HHS / United States
HHSN272201000054C / AI / NIAID NIH HHS / United States