Structure and function analysis of the essential 3′X domain of hepatitis C virus
| Autor: | Cristina Romero-López, Isaías Sanmartín, Jesús Castillo-Martínez, Alfredo Berzal-Herranz, Tamara Ovejero, José M. Gallego, Elisa Oltra, Beatriz Berzal-Herranz |
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| Přispěvatelé: | Ministerio de Economía y Competitividad (España), Universidad Católica de Valencia 'San Vicente Mártir' |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular RNA Folding Mutant Hepacivirus Biology Viral Nonstructural Proteins Virus Replication Article 03 medical and health sciences Humans Nucleotide Nucleic acid structure Molecular Biology 3' Untranslated Regions Base Pairing 030304 developmental biology Sequence (medicine) Palindromic sequence chemistry.chemical_classification 0303 health sciences Transition (genetics) Nucleotides 030302 biochemistry & molecular biology Inverted Repeat Sequences Translation (biology) Hepatitis C Cell biology Viral replication chemistry 3'X domain 5BSL3.2 domain hepatitis C virus RNA structure replication cycle Mutation Nucleic Acid Conformation RNA Viral Dimerization |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | The 3X domain of hepatitis C virus has been reported to control viral replication and translation by modulating the exposure of a nucleotide segment involved in a distal base-pairing interaction with an upstream 5BSL3.2 domain. To study the mechanism of this molecular switch, we have analyzed the structure of 3X mutants that favor one of the two previously proposed conformations comprising either two or three stem–loops. Only the two-stem conformation was found to be stable and to allow the establishment of the distal contact with 5BSL3.2, and also the formation of 3X domain homodimers by means of a universally conserved palindromic sequence. Nucleotide changes disturbing the two-stem conformation resulted in poorer replication and translation levels, explaining the high degree of conservation detected for this sequence. The switch function attributed to the 3X domain does not occur as a result of a transition between two- and three-stem conformations, but likely through the sequestration of the 5BSL3.2-binding sequence by formation of 3X homodimers. We thank R. Bartenschlager (University of Heidelberg, Germany) for providing the plasmid coding for HCV replicon pFK-I389-Fluc-NS3-3′ET, and Angel Cantero-Camacho for his contribution toward the design of mutant 3′X sequences. This work was supported by Ministerio de Economía y Competitividad of Spain (BFU-2012-30770 and BFU2015-65103-R to J.G., and BFU2015-64359-P to A.B.-H.) and Universidad Católica de Valencia of Spain (PRUCV/2015/629 and 2017-114-001 to J.G., and EDUCV contract E-46-2017-0118740 to J.C.-M.). |
| Databáze: | OpenAIRE |
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