Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

A high throughput RNA displacement assay for screening SARS-CoV-2 nsp10-nsp16 complex towards developing therapeutics for COVID-19

Sumera Perveen, Aliakbar Khalili Yazdi, Kanchan Devkota, Fengling Li, Pegah Ghiabi, Taraneh Hajian, Peter Loppnau, Albina Bolotokova, View ORCID ProfileMasoud Vedadi
doi: https://doi.org/10.1101/2020.10.14.340034
Sumera Perveen
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Aliakbar Khalili Yazdi
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kanchan Devkota
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Fengling Li
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pegah Ghiabi
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Taraneh Hajian
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Peter Loppnau
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Albina Bolotokova
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Masoud Vedadi
1Structural Genomics Consortium, University of Toronto, Toronto, Ontario, M5G 1L7, Canada
2Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, M5S 1A8, Canada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Masoud Vedadi
  • For correspondence: m.vedadi@utoronto.ca
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

Abstract

SARS-CoV-2, the coronavirus that causes COVID-19, evades the human immune system by capping its RNA. This process protects the viral RNA and is essential for its replication. Multiple viral proteins are involved in this RNA capping process including the nonstructural protein 16 (nsp16) which is an S-adenosyl-L-methionine (SAM)-dependent 2’-O-methyltransferase. Nsp16 is significantly active when in complex with another nonstructural protein, nsp10, which plays a key role in its stability and activity. Here we report the development of a fluorescence polarization (FP)-based RNA displacement assay for nsp10-nsp16 complex in 384-well format with a Z′-Factor of 0.6, suitable for high throughput screening. In this process, we purified the nsp10-nsp16 complex to higher than 95% purity and confirmed its binding to the methyl donor SAM, product of the reaction, SAH, and a common methyltransferase inhibitor, sinefungin using Isothermal Titration Calorimetry (ITC). The assay was further validated by screening a library of 1124 drug-like compounds. This assay provides a cost-effective high throughput method for screening nsp10-nsp16 complex for RNA-competitive inhibitors towards developing COVID-19 therapeutics.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted October 15, 2020.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
A high throughput RNA displacement assay for screening SARS-CoV-2 nsp10-nsp16 complex towards developing therapeutics for COVID-19
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
A high throughput RNA displacement assay for screening SARS-CoV-2 nsp10-nsp16 complex towards developing therapeutics for COVID-19
Sumera Perveen, Aliakbar Khalili Yazdi, Kanchan Devkota, Fengling Li, Pegah Ghiabi, Taraneh Hajian, Peter Loppnau, Albina Bolotokova, Masoud Vedadi
bioRxiv 2020.10.14.340034; doi: https://doi.org/10.1101/2020.10.14.340034
Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
A high throughput RNA displacement assay for screening SARS-CoV-2 nsp10-nsp16 complex towards developing therapeutics for COVID-19
Sumera Perveen, Aliakbar Khalili Yazdi, Kanchan Devkota, Fengling Li, Pegah Ghiabi, Taraneh Hajian, Peter Loppnau, Albina Bolotokova, Masoud Vedadi
bioRxiv 2020.10.14.340034; doi: https://doi.org/10.1101/2020.10.14.340034

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (2409)
  • Biochemistry (4757)
  • Bioengineering (3300)
  • Bioinformatics (14584)
  • Biophysics (6591)
  • Cancer Biology (5132)
  • Cell Biology (7383)
  • Clinical Trials (138)
  • Developmental Biology (4327)
  • Ecology (6826)
  • Epidemiology (2057)
  • Evolutionary Biology (9842)
  • Genetics (7308)
  • Genomics (9470)
  • Immunology (4509)
  • Microbiology (12595)
  • Molecular Biology (4903)
  • Neuroscience (28110)
  • Paleontology (198)
  • Pathology (799)
  • Pharmacology and Toxicology (1372)
  • Physiology (1996)
  • Plant Biology (4452)
  • Scientific Communication and Education (970)
  • Synthetic Biology (1293)
  • Systems Biology (3894)
  • Zoology (718)