Genomic RNA Elements Drive Phase Separation of the SARS-CoV-2 Nucleocapsid
| Autor: | Ethan J. Fritch, Christine A. Roden, Manolis Kellis, Amy S. Gladfelter, Mark A. Boerneke, Ralph S. Baric, Timothy P. Sheahan, Kevin M. Weeks, Grace A. McLaughlin, Chandra L. Theesfeld, Joanne Ekena, Yixuan J. Hou, Christiane Iserman, Chase A. Weidmann, Rachel Sealfon, Olga G. Troyanskaya, Irwin Jungreis |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Drug Evaluation
Preclinical Genome Viral Biology medicine.disease_cause Antiviral Agents Genome Article 03 medical and health sciences 0302 clinical medicine Chlorocebus aethiops medicine Animals Coronavirus Nucleocapsid Proteins Humans Nucleic acid structure Binding site Nucleocapsid Vero Cells Molecular Biology 030304 developmental biology Coronavirus 0303 health sciences SARS-CoV-2 HEK 293 cells COVID-19 RNA Cell Biology Phosphoproteins Small molecule COVID-19 Drug Treatment HEK293 Cells Biophysics Vero cell RNA Viral 030217 neurology & neurosurgery |
| Zdroj: | Molecular Cell |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2020.11.041 |
| Popis: | 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 with single-stranded RNA flanked by structure 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 presents a screenable process for identifying antiviral compounds effective against SARS-CoV-2. Graphical Abstract Highlights Phase separation occurs with the viral genome (gRNA) and at human body temperature. Phase separation is driven by specific elements in gRNA. RBD mutant N-protein fails to undergo LLPS; exhibits altered RNA crosslinking. N-protein forms liquid-like droplets in cells. Iserman and Roden et al. demonstrate phase separation (LLPS) of SARS-CoV-2 nucleocapsid (N-protein) with viral RNA. Viral RNA sequences promote or oppose phase separation depending on binding patterns of N-protein and genomic RNA. LLPS-promoting sequences occur at 5′ and 3′-Ends of the genome, suggestive of a genome packaging role. |
| Databáze: | OpenAIRE |
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