title: Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography.
abstract: Ebola virus is a highly pathogenic filovirus causing severe hemorrhagic fever with high mortality rates. It assembles heterogenous, filamentous, enveloped virus particles containing a negative-sense, single-stranded RNA genome packaged within a helical nucleocapsid (NC). We have used cryo-electron microscopy and tomography to visualize Ebola virus particles, as well as Ebola virus-like particles, in three dimensions in a near-native state. The NC within the virion forms a left-handed helix with an inner nucleoprotein layer decorated with protruding arms composed of VP24 and VP35. A comparison with the closely related Marburg virus shows that the N-terminal region of nucleoprotein defines the inner diameter of the Ebola virus NC, whereas the RNA genome defines its length. Binding of the nucleoprotein to RNA can assemble a loosely coiled NC-like structure; the loose coil can be condensed by binding of the viral matrix protein VP40 to the C terminus of the nucleoprotein, and rigidified by binding of VP24 and VP35 to alternate copies of the nucleoprotein. Four proteins (NP, VP24, VP35, and VP40) are necessary and sufficient to mediate assembly of an NC with structure, symmetry, variability, and flexibility indistinguishable from that in Ebola virus particles released from infected cells. Together these data provide a structural and architectural description of Ebola virus and define the roles of viral proteins in its structure and assembly.
authors.affiliation: Structural and Computational Biology Unit,  European Molecular Biology Laboratory, 69117 Heidelberg,  Germany.
authors.forename: Tanmay A M
authors.initials: TA
authors.lastname: Bharat
authors.forename: Takeshi
authors.initials: T
authors.lastname: Noda
authors.forename: James D
authors.initials: JD
authors.lastname: Riches
authors.forename: Verena
authors.initials: V
authors.lastname: Kraehling
authors.forename: Larissa
authors.initials: L
authors.lastname: Kolesnikova
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Kawaoka
authors.forename: John A G
authors.initials: JA
authors.lastname: Briggs
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods*; D029043:Ebolavirus / Q000648:ultrastructure*; D055032:Electron Microscope Tomography / Q000379:methods*; D016679:Genome, Viral / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D019251:Nucleocapsid / Q000235:genetics / Q000648:ultrastructure; D014771:Virion / Q000235:genetics / Q000648:ultrastructure