Work Description

Title: Raw data for "A narrow ratio of nucleic acid to SARS-CoV-2 N-protein enables phase separation" Open Access Deposited

http://creativecommons.org/licenses/by-nc/4.0/
Attribute Value
Abstract
  • SARS-CoV-2 Nucleocapsid protein (N) is a viral structural protein that packages the 30kb genomic RNA inside virions and forms condensates within infected cells through liquid-liquid phase separation (LLPS). N, in both soluble and condensed forms, has accessory roles in the viral life cycle including genome replication and immunosuppression. The ability to perform these tasks depends on phase separation and its reversibility. The conditions that stabilize and destabilize N condensates and the role of N-N interactions are poorly understood. We have investigated LLPS formation and dissolution in a minimalist system comprised of N protein and an ssDNA oligomer just long enough to support assembly. The short oligo allows us to focus on the role of N-N interaction. We have developed a sensitive FRET assay to interrogate LLPS assembly reactions from the perspective of the oligonucleotide. We find that N alone can form oligomers but that oligonucleotide enables their assembly into a three-dimensional phase. At a ~1:1
Methodology
  • The methods used to generate this data are fluorescence spectroscopy, fluorescence resonance energy transfer, absorbance, differential interference contrast microscopy, fluorescence microscopy, crosslinking, and SDS-PAGE.
Description
  • Spectroscopy data were collected on a BioTek Synergy H1 plate reader, then exported to and analyzed in Excel. 3D charts used in the manuscript were generated with OriginPro. Microscopy images are in proprietary .nd2 format of Nikon Elements software. .nd2 files were opened and processed with ImageJ. Gels were imaged on a BioRad Chemidoc.
Creator
Depositor
Contact information
Funding agency
  • National Institutes of Health (NIH)
  • Other Funding Agency
Other Funding agency
  • Indiana Clinical and Translational Sciences Institute
Keyword
Date coverage
  • 2022-12-21 to 2024-06-18
Citations to related material
  • Laughlin, P. M.; Young, K.; Gonzalez-Guiterrez, G.; Wang, J. C.-Y.,; Zlotnick, A. A narrow ratio of nucleic acid to SARS-CoV-2 N-protein enables phase separation. 2024.
Publisher
Resource type
Last modified
  • 09/09/2024
Location
  • Indiana University Bloomington
Coordinates
  • 39.165935826942714, -86.52245899494682
DOI
License
To Cite this Work:
Laughlin, P. Raw data for "A narrow ratio of nucleic acid to SARS-CoV-2 N-protein enables phase separation" [Data set]. Indiana University - DataCORE. https://doi.org/10.5967/p5mj-n362

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