@article{1a95ca108c3e42529a7741b9f6f63144,
title = "Genomic RNA Elements Drive Phase Separation of the SARS-CoV-2 Nucleocapsid",
abstract = "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.",
keywords = "SARS-CoV-2, Condensation, phase separation, packaging, RNP-MaP, RNA structure, nucleocapsid, coronavirus",
author = "Christiane Iserman and Roden, {Christine A.} and Boerneke, {Mark A.} and Sealfon, {Rachel S.G.} and McLaughlin, {Grace A.} and Irwin Jungreis and Fritch, {Ethan J.} and Hou, {Yixuan J.} and Joanne Ekena and Weidmann, {Chase A.} and Theesfeld, {Chandra L.} and Manolis Kellis and Troyanskaya, {Olga G.} and Baric, {Ralph S.} and Sheahan, {Timothy P.} and Weeks, {Kevin M.} and Gladfelter, {Amy S.}",
note = "Funding Information: We thank Rick Young, Phil Sharp, Alex Holehouse, Kathleen Hall, Andrea Sorrano, Ahmet Yildez, and their lab members for sharing data and discussions; Timothy Mitchison for discussions and critical reading of the manuscript; David Adalsteinsson for his help with ImageTank software; Ian Seim for analysis consultation and discussions; Benjamin Stormo for critical reading of the manuscript; Alain Laederach for initial discussion on genomic sequence; and James Iserman for essential logistical support. A.S.G. C.I. and C.A.R. were supported by NIH R01GM081506, Fast Grants Award #2139, and an HHMI faculty Scholar Award; C.A.R. was supported by NIH T32 CA 9156-43 and F32GM136164 and L'OREAL USA for Women in Science Fellowship. The work by R.S.G.S. A.S.B. and C.L.T. is supported by NIH grants R01HG005998, U54HL117798, and R01GM071966, HHS grant HHSN272201000054C, and Simons Foundation grant 395506 to O.G.T. K.M.W. M.A.B. and C.A.W. were supported by NIH (R35 GM122532 to K.M.W.). M.A.B. was supported by a Ruth L. Kirschstein Postdoctoral Fellowship (F32 GM128330). T.P.S. E.J.F. and R.S.B. were supported by National Institute of Allergy and Infectious Diseases grants (1U19AI142759; Antiviral Drug Discovery and Development Center). This project was supported in part by the North Carolina Policy Collaboratory at University of North Carolina at Chapel Hill with funding from the North Carolina Coronavirus Relief Fund established and appropriated by the North Carolina General Assembly. C.A.R. and C.I. contributed equally to this work. Authorship was determined alphabetically. C.A.R. C.I. and A.S.G. conceptualized the project, designed experiments, prepared figures, and drafted and edited the manuscript. C.A.R. and C.I. also performed experiments and analyzed data; M.A.B. designed and performed experiments and computational analyses, analyzed data, prepared figures, and wrote the manuscript. K.M.W. designed experiments, analyzed data, and wrote the manuscript; G.A.M. designed and performed experiments, analyzed data, edited the manuscript, and performed computational analyses; R.S.G.S. designed and performed computational analysis and edited the manuscript; I.J. supported computational analyses; M.K. supported I.J. C.L.T. provided fruitful discussion and edited the manuscript; O.G.T. provided support for R.S.G.S. and C.L.T.;. J.E. generated mutant plasmids; C.A.W. designed experiments and provided fruitful discussion; E.J.F. Y.J.H. T.P.S. and R.S.B. harvested viral RNA. K.M.W. is an advisor to and holds equity in Ribometrix, to which mutational profiling (MaP) technologies have been licensed. A.S.G. is a scientific advisor of Dewpoint Therapeutics. C.I. is currently employed at Dewpoint Therapeutics. All other authors declare that they have no competing interests. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = dec,
day = "17",
doi = "10.1016/j.molcel.2020.11.041",
language = "English (US)",
volume = "80",
pages = "1078--1091.e6",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "6",
}