Inhibition of hepatitis C virus (HCV) RNA polymerase by DNA aptamers: mechanism of inhibition of in vitro RNA synthesis and effect on HCV-infected cells
| Autor: | Thérèse Astier-Gin, Julie Rumi, Laura Tarrago-Litvak, Michel Ventura, Marie-Line Andreola, Christian Cazenave, Pantxika Bellecave, Ophélie Cosnefroy, Cathy Staedel |
|---|---|
| Přispěvatelé: | Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2008 |
| Předmět: |
MESH: SELEX Aptamer Technique
MESH: Aptamers Nucleotide Hepacivirus Viral Nonstructural Proteins Virus Replication medicine.disease_cause chemistry.chemical_compound RNA polymerase Pharmacology (medical) MESH: Hepacivirus Polymerase 0303 health sciences biology SELEX Aptamer Technique 030302 biochemistry & molecular biology virus diseases Aptamers Nucleotide 3. Good health Infectious Diseases [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology MESH: RNA Viral RNA Viral MESH: Virion MESH: RNA Replicase MESH: Cell Line Tumor Hepatitis C virus Aptamer RNA-dependent RNA polymerase Transfection Antiviral Agents Cell Line 03 medical and health sciences Cell Line Tumor medicine Humans NS5B 030304 developmental biology Pharmacology MESH: Humans MESH: Transfection MESH: Virus Replication Virion RNA RNA-Dependent RNA Polymerase Virology Molecular biology digestive system diseases MESH: Cell Line chemistry biology.protein MESH: Viral Nonstructural Proteins |
| Zdroj: | Antimicrobial Agents and Chemotherapy Antimicrobial Agents and Chemotherapy, American Society for Microbiology, 2008, 52 (6), pp.2097-110. ⟨10.1128/AAC.01227-07⟩ |
| ISSN: | 0066-4804 1098-6596 |
| DOI: | 10.1128/AAC.01227-07⟩ |
| Popis: | We describe here the further characterization of two DNA aptamers that specifically bind to hepatitis C virus (HCV) RNA polymerase (NS5B) and inhibit its polymerase activity in vitro. Although they were obtained from the same selection procedure and contain an 11-nucleotide consensus sequence, our results indicate that aptamers 27v and 127v use different mechanisms to inhibit HCV polymerase. While aptamer 27v was able to compete with the RNA template for binding to the enzyme and blocked both the initiation and the elongation of RNA synthesis, aptamer 127v competed poorly and exclusively inhibited initiation and postinitiation events. These results illustrate the power of the selective evolution of ligands by exponential enrichment in vitro selection procedure approach to select specific short DNA aptamers able to inhibit HCV NS5B by different mechanisms. We also determined that, in addition to an in vitro inhibitory effect on RNA synthesis, aptamer 27v was able to interfere with the multiplication of HCV JFH1 in Huh7 cells. The efficient cellular entry of these short DNAs and the inhibitory effect observed on human cells infected with HCV indicate that aptamers are useful tools for the study of HCV RNA synthesis, and their use should become a very attractive and alternative approach to therapy for HCV infection. |
| Databáze: | OpenAIRE |
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