title: Structural Basis of Vesicle Formation at the Inner Nuclear Membrane.
abstract: Vesicular nucleo-cytoplasmic transport is becoming recognized as a general cellular mechanism for translocation of large cargoes across the nuclear envelope. Cargo is recruited, enveloped at the inner nuclear membrane (INM), and delivered by membrane fusion at the outer nuclear membrane. To understand the structural underpinning for this trafficking, we investigated nuclear egress of progeny herpesvirus capsids where capsid envelopment is mediated by two viral proteins, forming the nuclear egress complex (NEC). Using a multi-modal imaging approach, we visualized the NEC in situ forming coated vesicles of defined size. Cellular electron cryo-tomography revealed a protein layer showing two distinct hexagonal lattices at its membrane-proximal and membrane-distant faces, respectively. NEC coat architecture was determined by combining this information with integrative modeling using small-angle X-ray scattering data. The molecular arrangement of the NEC establishes the basic mechanism for budding and scission of tailored vesicles at the INM.
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Christoph
authors.initials: C
authors.lastname: Hagen
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK; Diamond Light Source Ltd., Harwell Science and Innovation Campus, Didcot OX11 0DE, UK.
authors.forename: Kyle C
authors.initials: KC
authors.lastname: Dent
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Tzviya
authors.initials: T
authors.lastname: Zeev-Ben-Mordehai
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Michael
authors.initials: M
authors.lastname: Grange
authors.affiliation: Department of Molecular Biology, Princeton Neuroscience Institute, Princeton University, Washington Road, Princeton, NJ 08544, USA.
authors.forename: Jens B
authors.initials: JB
authors.lastname: Bosse
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Cathy
authors.initials: C
authors.lastname: Whittle
authors.affiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Barbara G
authors.initials: BG
authors.lastname: Klupp
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: C Alistair
authors.initials: CA
authors.lastname: Siebert
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Daven
authors.initials: D
authors.lastname: Vasishtan
authors.affiliation: Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Felix J B
authors.initials: FJ
authors.lastname: Bäuerlein
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
authors.forename: Juliana
authors.initials: J
authors.lastname: Cheleski
authors.affiliation: Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, 12489 Berlin, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Werner
authors.affiliation: Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, 12489 Berlin, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Guttmann
authors.affiliation: Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, 12489 Berlin, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Rehbein
authors.affiliation: Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, 12489 Berlin, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Henzler
authors.affiliation: Micron Oxford, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
authors.forename: Justin
authors.initials: J
authors.lastname: Demmerle
authors.affiliation: Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität München, Pettenkoferstr. 9a, 80336 Munich, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Adler
authors.affiliation: Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität München, Pettenkoferstr. 9a, 80336 Munich, Germany.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Koszinowski
authors.affiliation: Micron Oxford, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
authors.forename: Lothar
authors.initials: L
authors.lastname: Schermelleh
authors.affiliation: Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, 12489 Berlin, Germany.
authors.forename: Gerd
authors.initials: G
authors.lastname: Schneider
authors.affiliation: Department of Molecular Biology, Princeton Neuroscience Institute, Princeton University, Washington Road, Princeton, NJ 08544, USA.
authors.forename: Lynn W
authors.initials: LW
authors.lastname: Enquist
authors.affiliation: Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Jürgen M
authors.initials: JM
authors.lastname: Plitzko
authors.affiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany. Electronic address: thomas.mettenleiter@fli.bund.de.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
authors.affiliation: Oxford Particle Imaging Centre, Division of Structural Biology, Wellcome Trust Centre for Human Genetics, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK. Electronic address: kay@strubi.ox.ac.uk.
authors.forename: Kay
authors.initials: K
authors.lastname: Grünewald
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D002213:Capsid / Q000378:metabolism* / Q000648:ultrastructure; D002522:Chlorocebus aethiops; D020285:Cryoelectron Microscopy; D055032:Electron Microscope Tomography; D018259:Herpesvirus 1, Human / Q000378:metabolism; D011558:Herpesvirus 1, Suid / Q000378:metabolism; D009685:Nuclear Envelope / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D011740:Pyrimidine Dimers; D053838:Scattering, Small Angle; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure*; D014709:Vero Cells; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D009687:Nuclear Proteins; D011740:Pyrimidine Dimers; C108721:UL31 protein, Human herpesvirus 1; C403420:UL34 protein, Human herpesvirus 1; D014764:Viral Proteins