Local RNA flexibility perturbation of the IRES element induced by a novel ligand inhibits viral RNA translation
| Autor: | Alejandro Trapote, Jorge Ramajo, Encarnación Martínez-Salas, Anna Grandas, Gloria Lozano, Jordi Robles, Xavier Elduque, Enrique Pedroso |
|---|---|
| Přispěvatelé: | Ministerio de Economía y Competitividad (España), Fundación Ramón Areces |
| Rok vydání: | 2015 |
| Předmět: |
Picornavirus
Molecular Sequence Data RNA ligands RNA-dependent RNA polymerase Genome Viral Internal Ribosome Entry Sites Ligands antiviral molecules translation initiation Fluorescence Eukaryotic translation Protein biosynthesis Nucleic acid structure RNA structure Molecular Biology biology Base Sequence Cell-Free System Hydroxyl Radical 2-aminobenzimidazole fungi RNA Cell Biology biology.organism_classification Molecular biology Research Papers Cell biology fluorescence binding assay Internal ribosome entry site picornavirus IRES elements Interaction with host Foot-and-Mouth Disease Virus Protein Biosynthesis Solvents Nucleic Acid Conformation RNA Viral Benzimidazoles SHAPE probing |
| Zdroj: | RNA Biology Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1555-8584 |
| Popis: | The internal ribosome entry site (IRES) element located at the 5’untranslated genomic region of various RNA viruses mediates cap-independent initiation of translation. Picornavirus IRES activity is highly dependent on both its structural organization and its interaction with host factors. Small molecules able to interfere with RNA function are valuable candidates for antiviral agents. Here we show that a small molecule based on benzimidazole (IRAB) inhibited foot-andmouth disease virus (FMDV) IRES-dependent protein synthesis in cells transfected with infectious RNA leading to a decrease of the virus titer, which was higher than that induced by a structurally related benzimidazole derivative. Interestingly, IRAB preferentially inhibited IRES-dependent translation in cell free systems in a dose-dependent manner. RNA structural analysis by SHAPE demonstrated an increased local flexibility of the IRES structure upon incubation with IRAB, which affected 3 stem-loops (SL) of domain 3. Fluorescence binding assays conducted with individual aminopurinelabeled oligoribonucleotides indicated that the SL3A binds IRAB (EC 18 μM). Taken together, the results derived from SHAPE reactivity and fluorescence binding assays suggested that the target site of IRAB within the FMDV IRES might be a folded RNA structure that involves the entire apical region of domain 3. Our data suggest that the conformational changes induced by this compound on a specific region of the IRES structure which is essential for its activity is, at least in part, responsible for the reduced IRES efficiency observed in cell free lysates and, particularly, in RNA-transfected cells. Ministerio de Economia y Competitividad (MINECO), and by an Institutional grant from Fundación Ramón Areces |
| Databáze: | OpenAIRE |
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