Abulwerdi, F, Fatehi, F, Manfield, I orcid.org/0000-0003-3765-0325 et al. (5 more authors) (2022) Dataset Of High-Throughput Ligand Screening Against the RNA Packaging Signals Regulating Hepatitis B Virus Nucleocapsid Formation. Data in Brief, 42. 108206. p. 108206. ISSN 2352-3409
Abstract
Multiple ssRNA viruses which infect bacteria, plants or humans use RNA Packaging Signal (PS)-mediated regulation during assembly to package their genomes faithfully and efficiently [1], [2], [3], [4], [5]. PSs typically comprise short nucleotide recognition motifs, most often presented in the unpaired region of RNA stem-loops, and often bind their cognate coat proteins (CPs) with nanomolar affinity. PSs identified to date are resilient in the face of the typical error prone replication of their virus-coded polymerases, making them potential drug targets [1], [2], [3]. An immobilised array of small molecular weight, drug-like compounds was panned [6] against a fluorescently-labelled oligonucleotide encompassing the most conserved Hepatitis B Virus (HBV) PS, PS1 [3], known to be a major determinant in nucleocapsid formation [7]. This identified > 70 compounds that bind PS1 uniquely in the array. The commercially available 66 of these were tested for their potential effect(s) on HBV nucleocapsid-like particle (NCP) assembly in vitro [8], which identified potent assembly inhibitors. Here, we describe a high-throughput screen for such effects using employing fluorescence anisotropy in a 96-well microplate format. HBV genomic RNAs (gRNA) and short oligonucleotides encompassing PS1 were 5ʹ labelled with an Alexa Fluor 488 dye. Excess (with respect to stoichiometric T=4 NCP formation [9]) HBV core protein (Cp) dimers were titrated robotically into solutions containing each of these RNAs stepwise, using a Biomek 4000 liquid handling robot. The anisotropy values of these mixtures were monitored using a POLARstar microplate reader. NCP-like structures were challenged with RNase A to identify reactions that did not result in complete NCP formation [10]. The results imply that ∼50 % of the compounds prevent complete NCP formation, highlighting both PS-meditated assembly and the PS-binding compounds as potential directly-acting anti-virals with a novel molecular target. Importantly, this method allows high-throughput in vitro screening for assembly inhibitors in this major human pathogen.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Keywords: | Hepatitis B Virus nucleocapsid assembly; RNA Packaging Signal-mediated virus assembly; high-throughput screening; assembly inhibitors |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Biological Chemistry (Leeds) |
Funding Information: | Funder Grant number Medical Research Foundation MRF-044-0002-RG-PATEL |
Depositing User: | Symplectic Publications |
Date Deposited: | 28 Apr 2022 09:52 |
Last Modified: | 29 Jul 2022 13:40 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.dib.2022.108206 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:186167 |