Estudio de los complejos replicativos del torovirus equino Bev

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 22-10-2015

Detalles Bibliográficos
Autor: Ávila Pérez, Ginés Francisco
Tipo de recurso: tesis doctoral
Fecha de publicación:2015
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:español
OAI Identifier:oai:repositorio.uam.es:10486/670313
Acceso en línea:http://hdl.handle.net/10486/670313
Access Level:acceso abierto
Palabra clave:Coronavirus - Reproducción - Tesis doctorales
Biología y Biomedicina / Biología
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dc.title.none.fl_str_mv Estudio de los complejos replicativos del torovirus equino Bev
title Estudio de los complejos replicativos del torovirus equino Bev
spellingShingle Estudio de los complejos replicativos del torovirus equino Bev
Ávila Pérez, Ginés Francisco
Coronavirus - Reproducción - Tesis doctorales
Biología y Biomedicina / Biología
title_short Estudio de los complejos replicativos del torovirus equino Bev
title_full Estudio de los complejos replicativos del torovirus equino Bev
title_fullStr Estudio de los complejos replicativos del torovirus equino Bev
title_full_unstemmed Estudio de los complejos replicativos del torovirus equino Bev
title_sort Estudio de los complejos replicativos del torovirus equino Bev
dc.creator.none.fl_str_mv Ávila Pérez, Ginés Francisco
author Ávila Pérez, Ginés Francisco
author_facet Ávila Pérez, Ginés Francisco
author_role author
dc.contributor.none.fl_str_mv Rodríguez Aguirre, Dolores
Departamento de Biología Molecular
Facultad de Ciencias
CSIC. Centro Nacional de Biotecnología (CNB)
dc.subject.none.fl_str_mv Coronavirus - Reproducción - Tesis doctorales
Biología y Biomedicina / Biología
topic Coronavirus - Reproducción - Tesis doctorales
Biología y Biomedicina / Biología
description Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 22-10-2015
publishDate 2015
dc.date.none.fl_str_mv 2015
2015-10-22
dc.type.none.fl_str_mv doctoral thesis
http://purl.org/coar/resource_type/c_db06
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.none.fl_str_mv http://hdl.handle.net/10486/670313
url http://hdl.handle.net/10486/670313
dc.language.none.fl_str_mv Español
spa
language_invalid_str_mv Español
language spa
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
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eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Biblos-e Archivo. Repositorio Institucional de la UAM
instname:Universidad Autónoma de Madrid
instname_str Universidad Autónoma de Madrid
reponame_str Biblos-e Archivo. Repositorio Institucional de la UAM
collection Biblos-e Archivo. Repositorio Institucional de la UAM
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spelling Estudio de los complejos replicativos del torovirus equino BevÁvila Pérez, Ginés FranciscoCoronavirus - Reproducción - Tesis doctoralesBiología y Biomedicina / BiologíaTesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 22-10-2015Plus-stranded RNA viruses replicate in the cytosol of infected cells, in membrane-bound replication complexes containing the replicase proteins, the viral RNA and host proteins. The formation of the replication and transcription complexes (RTCs) through the rearrangement of cellular membranes is currently being actively studied for viruses belonging to different viral families. In this work we identified double membrane vesicles (DMVs) in the cytoplasm of cells infected with the equine torovirus Berne virus (BEV), the prototype member of the Torovirus genus (Coronaviridae Family, Nidovirales Order). Using confocal microscopy and transmission electron microscopy we observed a close relationship between the RTCs and the DMVs of BEV. The examination of BEV infected cells revealed that the replicase proteins colocalized with each other and with newly synthetized RNA, and were associated to the membrane rearrangement induced by BEV. However, the dsRNA, an intermediate of viral replication, was exclusively limited to the interior of DMVs. Also, an in-depth ultrastructural analysis of cells infected with BEV was performed to characterize the architecture of torovirus replication factories and to learn about their biogenesis during the infection. Analysis by conventional transmission electron microscopy suggested that the DMVs form a reticulovesicular network (RVN) resembling those described for the related severe acute respiratory syndrome coronavirus (SARS-CoV) and the equine arteritis virus (EAV). Using serial sectioning and electron tomography we confirmed the formation of a RVN in BEV infected cells, where the DMVs outer membranes are interconnected with each other and with the ER. Like in EAV, convoluted membranes were not observed in the RVNs. However, we observed paired membranes lacking luminal space, which are connected with the DMVs, and likely represent early structures that will evolve to give rise to DMVs. Interestingly, curled membranes resembling the spherules described in IBV were observed at late time post-infection in BEV-infected cells, however, after careful examination of the tomograms we concluded that they are not true spherules, but aberrant structures arising late on the DMV formation. This work complements the knowledge about the membrane rearrangements promoted by nidovirus infection, and all this taken together indicates that the nidoviruses use a similar strategy to establish their membranous replication factories. On the other hand, recent studies suggest that some positive-stranded RNA viruses exploit the autophagy pathway to facilitate their own replication. We demonstrated that BEV induces autophagy, with an active flux, in the infected cells at late times post-infection. However, the autophagy induction by nutrient deprivation, or its inhibition by pharmacological treatment with inhibitors of autophagosome formation or agents that block the autophagosome degradation, does not produce changes in BEV replication. In the same way, the replication of BEV was not affected in autophagy deficient cells lacking Beclin-1 or LC3B. Overall, our results suggest that the autophagy pathway is activated during BEV infection at late time post-infection, but this activation is not necessary for the viral replicationRodríguez Aguirre, DoloresDepartamento de Biología MolecularFacultad de CienciasCSIC. Centro Nacional de Biotecnología (CNB)20152015-10-22doctoral thesishttp://purl.org/coar/resource_type/c_db06NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/doctoralThesisapplication/pdfhttp://hdl.handle.net/10486/670313reponame:Biblos-e Archivo. Repositorio Institucional de la UAMinstname:Universidad Autónoma de MadridEspañolspaopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:repositorio.uam.es:10486/6703132026-06-23T12:46:27Z
score 15.300719

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