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Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures
Hepatitis C virus (HCV) is a major cause of liver-related diseases and hepatocellular carcinoma. The helicase domain of one of the nonstructural proteins of HCV, NS3 (nonstructural protein 3), is essential for viral replication; however, its specific biological role is still under investigation. Her...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9582721/ https://www.ncbi.nlm.nih.gov/pubmed/36108740 http://dx.doi.org/10.1016/j.jbc.2022.102486 |
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author | Belachew, Binyam Gao, Jun Byrd, Alicia K. Raney, Kevin D. |
author_facet | Belachew, Binyam Gao, Jun Byrd, Alicia K. Raney, Kevin D. |
author_sort | Belachew, Binyam |
collection | PubMed |
description | Hepatitis C virus (HCV) is a major cause of liver-related diseases and hepatocellular carcinoma. The helicase domain of one of the nonstructural proteins of HCV, NS3 (nonstructural protein 3), is essential for viral replication; however, its specific biological role is still under investigation. Here, we set out to determine the interaction between a purified recombinant full length NS3 and synthetic guanine-rich substrates that represent the conserved G-quadruplex (G4)-forming sequences in the HCV-positive and HCV-negative strands. We performed fluorescence anisotropy binding, G4 reporter duplex unwinding, and G4RNA trapping assays to determine the binding and G4 unfolding activity of NS3. Our data suggest that NS3 can unfold the conserved G4 structures present within the genome and the negative strand of HCV. Additionally, we found the activity of NS3 on a G4RNA was reduced significantly in the presence of a G4 ligand. The ability of NS3 to unfold HCV G4RNA could imply a novel biological role of the viral helicase in replication. |
format | Online Article Text |
id | pubmed-9582721 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-95827212022-10-21 Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures Belachew, Binyam Gao, Jun Byrd, Alicia K. Raney, Kevin D. J Biol Chem Research Article Hepatitis C virus (HCV) is a major cause of liver-related diseases and hepatocellular carcinoma. The helicase domain of one of the nonstructural proteins of HCV, NS3 (nonstructural protein 3), is essential for viral replication; however, its specific biological role is still under investigation. Here, we set out to determine the interaction between a purified recombinant full length NS3 and synthetic guanine-rich substrates that represent the conserved G-quadruplex (G4)-forming sequences in the HCV-positive and HCV-negative strands. We performed fluorescence anisotropy binding, G4 reporter duplex unwinding, and G4RNA trapping assays to determine the binding and G4 unfolding activity of NS3. Our data suggest that NS3 can unfold the conserved G4 structures present within the genome and the negative strand of HCV. Additionally, we found the activity of NS3 on a G4RNA was reduced significantly in the presence of a G4 ligand. The ability of NS3 to unfold HCV G4RNA could imply a novel biological role of the viral helicase in replication. American Society for Biochemistry and Molecular Biology 2022-09-13 /pmc/articles/PMC9582721/ /pubmed/36108740 http://dx.doi.org/10.1016/j.jbc.2022.102486 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Belachew, Binyam Gao, Jun Byrd, Alicia K. Raney, Kevin D. Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title | Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title_full | Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title_fullStr | Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title_full_unstemmed | Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title_short | Hepatitis C virus nonstructural protein NS3 unfolds viral G-quadruplex RNA structures |
title_sort | hepatitis c virus nonstructural protein ns3 unfolds viral g-quadruplex rna structures |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9582721/ https://www.ncbi.nlm.nih.gov/pubmed/36108740 http://dx.doi.org/10.1016/j.jbc.2022.102486 |
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