Por favor, use este identificador para citar o enlazar este Item:http://hdl.handle.net/20.500.12105/19990
Título
Generation of Replication-Proficient Influenza Virus NS1 Point Mutants with Interferon-Hyperinducer Phenotype
Autor(es)
Fecha de publicación
2014-06-02
Cita
Perez-Cidoncha M, Killip MJ, Asensio VJJ, Fernandez Y, Bengoechea JA, Randall RE, et al. Generation of Replication-Proficient Influenza Virus NS1 Point Mutants with Interferon-Hyperinducer Phenotype. PLoS One. 2014 Jun 02;9(6):e98668.
Idioma
Inglés
Tipo de documento
research article
Resumen
The NS1 protein of influenza A viruses is the dedicated viral interferon (IFN)-antagonist. Viruses lacking NS1 protein expression cannot multiply in normal cells but are viable in cells deficient in their ability to produce or respond to IFN. Here we report an unbiased mutagenesis approach to identify positions in the influenza A NS1 protein that modulate the IFN response upon infection. A random library of virus ribonucleoproteins containing circa 40 000 point mutants in NS1 were transferred to infectious virus and amplified in MDCK cells unable to respond to interferon. Viruses that activated the interferon (IFN) response were subsequently selected by their ability to induce expression of green-fluorescent protein (GFP) following infection of A549 cells bearing an IFN promoter-dependent GFP gene. Using this approach we isolated individual mutant viruses that replicate to high titers in IFN-compromised cells but, compared to wild type viruses, induced higher levels of IFN in IFN-competent cells and had a reduced capacity to counteract exogenous IFN. Most of these viruses contained not previously reported NS1 mutations within either the RNA-binding domain, the effector domain or the linker region between them. These results indicate that subtle alterations in NS1 can reduce its effectiveness as an IFN antagonist without affecting the intrinsic capacity of the virus to multiply. The general approach reported here may facilitate the generation of replication-proficient, IFN-inducing virus mutants, that potentially could be developed as attenuated vaccines against a variety of viruses.
MESH
Viral Nonstructural Proteins | Green Fluorescent Proteins | Base Sequence | Humans | Cell Line | Immunity, Innate | Promoter Regions, Genetic | Virus Replication | Dogs | Influenza A virus | Interferons | Animals | DNA Primers | Point Mutation
DECS
Animales | Cartilla de ADN | Replicación Viral | Inmunidad Innata | Perros | Línea Celular | Virus de la Influenza A | Interferones | Secuencia de Bases | Proteínas Fluorescentes Verdes | Humanos | Regiones Promotoras Genéticas | Proteínas no Estructurales Virales | Mutación Puntual
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DOI
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