title: Ultrastructural Characterization of Zika Virus Replication Factories.
abstract: A global concern has emerged with the pandemic spread of Zika virus (ZIKV) infections that can cause severe neurological symptoms in adults and newborns. ZIKV is a positive-strand RNA virus replicating in virus-induced membranous replication factories (RFs). Here we used various imaging techniques to investigate the ultrastructural details of ZIKV RFs and their relationship with host cell organelles. Analyses of human hepatic cells and neural progenitor cells infected with ZIKV revealed endoplasmic reticulum (ER) membrane invaginations containing pore-like openings toward the cytosol, reminiscent to RFs in Dengue virus-infected cells. Both the MR766 African strain and the H/PF/2013 Asian strain, the latter linked to neurological diseases, induce RFs of similar architecture. Importantly, ZIKV infection causes a drastic reorganization of microtubules and intermediate filaments forming cage-like structures surrounding the viral RF. Consistently, ZIKV replication is suppressed by cytoskeleton-targeting drugs. Thus, ZIKV RFs are tightly linked to rearrangements of the host cell cytoskeleton.
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Mirko
authors.initials: M
authors.lastname: Cortese
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Goellner
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Eliana Gisela
authors.initials: EG
authors.lastname: Acosta
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Christopher John
authors.initials: CJ
authors.lastname: Neufeldt
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Oleksiuk
authors.affiliation: Advanced Light Microscopy Facility, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Marko
authors.initials: M
authors.lastname: Lampe
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Uta
authors.initials: U
authors.lastname: Haselmann
authors.affiliation: Electron Microscopy Core Facility, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Funaya
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Schieber
authors.affiliation: Electron Microscopy Core Facility, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Paolo
authors.initials: P
authors.lastname: Ronchi
authors.affiliation: Electron Microscopy Core Facility, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Schorb
authors.affiliation: Department of Neurobiology, Interdisciplinary Center for Neurosciences (IZN), Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Priit
authors.initials: P
authors.lastname: Pruunsild
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany; Electron Microscopy Core Facility, European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Schwab
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany; Institut National de la Recherche Scientifique, Institut Armand-Frappier, Québec H7V 1B7, Canada.
authors.forename: Laurent
authors.initials: L
authors.lastname: Chatel-Chaix
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany.
authors.forename: Alessia
authors.initials: A
authors.lastname: Ruggieri
authors.affiliation: Department of Infectious Diseases, Molecular Virology, Heidelberg University, 69120 Heidelberg, Germany; German Center for Infection Research (DZIF), Heidelberg University, 69120 Heidelberg, Germany. Electronic address: ralf.bartenschlager@med.uni-heidelberg.de.
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000648:ultrastructure / Q000821:virology; D057809:HEK293 Cells; D022781:Hepatocytes / Q000648:ultrastructure / Q000821:virology; D054884:Host-Pathogen Interactions / Q000502:physiology*; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism / Q000648:ultrastructure; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D058953:Neural Stem Cells / Q000648:ultrastructure / Q000821:virology; D013234:Stem Cells / Q000648:ultrastructure / Q000821:virology; D014709:Vero Cells; D014779:Virus Replication / Q000502:physiology*; D000071244:Zika Virus / Q000378:metabolism / Q000648:ultrastructure*; D000071243:Zika Virus Infection / Q000378:metabolism / Q000821:virology*
keywords: Zika virus; electron microscopy; electron tomography; flavivirus; human neural progenitor cells; intermediate filaments; live-cell imaging; microtubules; replication factories; replication organelles