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Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures
Preprint   Open access

Double-stranded RNA drives SARS-CoV-2 nucleocapsid protein to undergo phase separation at specific temperatures

Christine A. Roden, Yifan Dai, Ian Seim, Myungwoon Lee, Rachel Sealfon, Grace A. McLaughlin, Mark A. Boerneke, Christiane Iserman, Samuel A. Wey, Joanne L. Ekena, …
bioRxiv
15 Jun 2021
DOI: https://doi.org/10.1101/2021.06.14.448452
PMID: 34159327
url
https://doi.org/10.1213/ANE.0b013e3182407c25View
Published, Version of Record (VoR)CC BY V4.0,  Open

Abstract

Betacoronavirus SARS-CoV-2 infections caused the global Covid-19 pandemic. The nucleocapsid protein (N-protein) is required for multiple steps in the betacoronavirus replication cycle. SARS-CoV-2-N-protein is known to undergo liquid-liquid phase separation (LLPS) with specific RNAs at particular temperatures to form condensates. We show that N-protein recognizes at least two separate and distinct RNA motifs, both of which require double-stranded RNA (dsRNA) for LLPS. These motifs are separately recognized by N-protein’s two RNA binding domains (RBDs). Addition of dsRNA accelerates and modifies N-protein LLPS in vitro and in cells and controls the temperature condensates form. The abundance of dsRNA tunes N-protein-mediated translational repression and may confer a switch from translation to genome packaging. Thus, N-protein’s two RBDs interact with separate dsRNA motifs, and these interactions impart distinct droplet properties that can support multiple viral functions. These experiments demonstrate a paradigm of how RNA structure can control the properties of biomolecular condensates.

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