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      Structural investigation of C6/36 and Vero cell cultures infected with a Brazilian Zika virus

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          Abstract

          Zika virus (ZIKV) is a member of the flavivirus genus, and its genome is approximately 10.8 kilobases of positive-strand RNA enclosed in a capsid and surrounded by a membrane. Studies on the replication dynamics of ZIKV are scarce, which limits the development of antiviral agents and vaccines directed against ZIKV. In this study, Aedes albopictus mosquito lineage cells (C6/36 cells) and African green monkey kidney epithelial cells (Vero cells) were inoculated with a ZIKV sample isolated from a Brazilian patient, and the infection was characterized by immunofluorescence staining, phase contrast light microscopy, transmission electron microscopy and real-time RT-PCR. The infection was observed in both cell lineages, and ZIKV particles were observed inside lysosomes, the rough endoplasmic reticulum and viroplasm-like structures. The susceptibility of C6/36 and Vero cells to ZIKV infection was demonstrated. Moreover, this study showed that part of the replicative cycle may occur within viroplasm-like structures, which has not been previously demonstrated in other flaviviruses.

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          Composition and Three-Dimensional Architecture of the Dengue Virus Replication and Assembly Sites

           Sonja Welsch,  Sven Miller,  Inés Romero-Brey … (2009)
          Summary Positive-strand RNA viruses are known to rearrange cellular membranes to facilitate viral genome replication. The biogenesis and three-dimensional organization of these membranes and the link between replication and virus assembly sites is not fully clear. Using electron microscopy, we find Dengue virus (DENV)-induced vesicles, convoluted membranes, and virus particles to be endoplasmic reticulum (ER)-derived, and we detect double-stranded RNA, a presumed marker of RNA replication, inside virus-induced vesicles. Electron tomography (ET) shows DENV-induced membrane structures to be part of one ER-derived network. Furthermore, ET reveals vesicle pores that could enable release of newly synthesized viral RNA and reveals budding of DENV particles on ER membranes directly apposed to vesicle pores. Thus, DENV modifies ER membrane structure to promote replication and efficient encapsidation of the genome into progeny virus. This architecture of DENV replication and assembly sites could explain the coordination of distinct steps of the flavivirus replication cycle.
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            Virus factories: associations of cell organelles for viral replication and morphogenesis

             Reyes R Novoa,  Gloria Calderita,  Rocío Arranz … (2005)
            Abstract Genome replication and assembly of viruses often takes place in specific intracellular compartments where viral components concentrate, thereby increasing the efficiency of the processes. For a number of viruses the formation of ‘factories’ has been described, which consist of perinuclear or cytoplasmic foci that mostly exclude host proteins and organelles but recruit specific cell organelles, building a unique structure. The formation of the viral factory involves a number of complex interactions and signalling events between viral and cell factors. Mitochondria, cytoplasmic membranes and cytoskeletal components frequently participate in the formation of viral factories, supplying basic and common needs for key steps in the viral replication cycle.
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              Autophagic machinery activated by dengue virus enhances virus replication

               Ying-Ray Lee,  Huan-Yao Lei,  Ming-Tao Liu … (2008)
              Autophagy is a cellular response against stresses which include the infection of viruses and bacteria. We unravel that Dengue virus-2 (DV2) can trigger autophagic process in various infected cell lines demonstrated by GFP-LC3 dot formation and increased LC3-II formation. Autophagosome formation was also observed under the transmission electron microscope. DV2-induced autophagy further enhances the titers of extracellular and intracellular viruses indicating that autophagy can promote viral replication in the infected cells. Moreover, our data show that ATG5 protein is required to execute DV2-induced autophagy. All together, we are the first to demonstrate that DV can activate autophagic machinery that is favorable for viral replication.
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                Author and article information

                Contributors
                Debora Ferreira Barreto-Vieira:
                ORCID: http://orcid.org/0000-0003-0094-9999
                Role: ConceptualizationRole: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: Writing – original draftRole: Writing – review & editing
                Fernanda Cunha Jácome: Role: Methodology
                Marcos Alexandre Nunes da Silva: Role: Methodology
                Gabriela Cardoso Caldas: Role: Methodology
                Ana Maria Bispo de Filippis: Role: Methodology
                Patrícia Carvalho de Sequeira: Role: Methodology
                Elen Mello de Souza: Role: Methodology
                Audrien Alves Andrade: Role: Methodology
                Pedro Paulo de Abreu Manso: Role: Methodology
                Gisela Freitas Trindade: Role: Methodology
                Sheila Maria Barbosa Lima: Role: Methodology
                Ortrud Monika Barth: Role: Resources
                Zheng Xing: Role: Editor
                Journal
                Journal ID (nlm-ta): PLoS One
                Journal ID (iso-abbrev): PLoS ONE
                Journal ID (publisher-id): plos
                Journal ID (pmc): plosone
                Title: PLoS ONE
                Publisher: Public Library of Science (San Francisco, CA USA )
                ISSN (Electronic): 1932-6203
                Publication date (Electronic): 12 September 2017
                Publication date Collection: 2017
                Volume: 12
                Issue: 9
                Affiliations
                [1 ] Laboratory of Morphology and Viral Morphogenesis, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil, Rio de Janeiro, RJ, Brazil
                [2 ] Laboratory of Flavivirus, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil, Rio de Janeiro, RJ, Brazil
                [3 ] Laboratory of Pathology, Instituto Oswaldo Cruz, Fiocruz, Avenida Brasil, Rio de Janeiro, RJ, Brazil
                [4 ] Laboratory of Virological Technology, Bio-Manguinhos, Avenida Brasil, Rio de Janeiro, RJ, Brazil
                University of Minnesota College of Veterinary Medicine, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Article
                Publisher ID: PONE-D-17-15841
                DOI: 10.1371/journal.pone.0184397
                PMC ID: 5595330
                PubMed ID: 28898286
                Copyright statement: © 2017 Barreto-Vieira et al
                License:

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 10, Tables: 1, Pages: 18
                Product
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100004586, Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro;
                Award Recipient :
                ORCID: http://orcid.org/0000-0003-0094-9999
                This study was funded by Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Faperj, and Ana Maria Bispo de Filippis is supported by the European Union's Horizon 2020 program under grant agreement No. 734857. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Subject: Research Article
                Subject: Biology and life sciences
                Subject: Organisms
                Subject: Viruses
                Subject: RNA viruses
                Subject: Flaviviruses
                Subject: Zika Virus
                Subject: Biology and Life Sciences
                Subject: Microbiology
                Subject: Medical Microbiology
                Subject: Microbial Pathogens
                Subject: Viral Pathogens
                Subject: Flaviviruses
                Subject: Zika Virus
                Subject: Medicine and Health Sciences
                Subject: Pathology and Laboratory Medicine
                Subject: Pathogens
                Subject: Microbial Pathogens
                Subject: Viral Pathogens
                Subject: Flaviviruses
                Subject: Zika Virus
                Subject: Biology and Life Sciences
                Subject: Organisms
                Subject: Viruses
                Subject: Viral Pathogens
                Subject: Flaviviruses
                Subject: Zika Virus
                Subject: Research and Analysis Methods
                Subject: Biological Cultures
                Subject: Cell Lines
                Subject: Vero Cells
                Subject: Biology and Life Sciences
                Subject: Microbiology
                Subject: Virology
                Subject: Viral Replication
                Subject: Biology and Life Sciences
                Subject: Cell Biology
                Subject: Cellular Structures and Organelles
                Subject: Endoplasmic Reticulum
                Subject: Biology and Life Sciences
                Subject: Cell Biology
                Subject: Cell Processes
                Subject: Secretory Pathway
                Subject: Endoplasmic Reticulum
                Subject: Biology and Life Sciences
                Subject: Cell Biology
                Subject: Cellular Structures and Organelles
                Subject: Endoplasmic Reticulum
                Subject: Rough Endoplasmic Reticulum
                Subject: Biology and Life Sciences
                Subject: Cell Biology
                Subject: Cell Processes
                Subject: Secretory Pathway
                Subject: Endoplasmic Reticulum
                Subject: Rough Endoplasmic Reticulum
                Subject: Biology and Life Sciences
                Subject: Microbiology
                Subject: Virology
                Subject: Viral Structure
                Subject: Research and Analysis Methods
                Subject: Microscopy
                Subject: Electron Microscopy
                Subject: Transmission Electron Microscopy
                Subject: Biology and Life Sciences
                Subject: Cell Biology
                Subject: Cellular Structures and Organelles
                Subject: Lysosomes
                Custom metadata
                Data Availability All relevant data are within the paper.

                Data availability:

                ScienceOpen disciplines: Uncategorized

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