TY - JOUR
T1 - Structural Basis of Vesicle Formation at the Inner Nuclear Membrane
AU - Hagen, Christoph
AU - Dent, Kyle C.
AU - Zeev-Ben-Mordehai, Tzviya
AU - Grange, Michael
AU - Bosse, Jens B.
AU - Whittle, Cathy
AU - Klupp, Barbara G.
AU - Siebert, C. Alistair
AU - Vasishtan, Daven
AU - Bäuerlein, Felix J.B.
AU - Cheleski, Juliana
AU - Werner, Stephan
AU - Guttmann, Peter
AU - Rehbein, Stefan
AU - Henzler, Katja
AU - Demmerle, Justin
AU - Adler, Barbara
AU - Koszinowski, Ulrich
AU - Schermelleh, Lothar
AU - Schneider, Gerd
AU - Enquist, Lynn W.
AU - Plitzko, Jürgen M.
AU - Mettenleiter, Thomas C.
AU - Grünewald, Kay
N1 - Funding Information:
We thank Wolfram Antonin (Tübingen) for critical discussion and Marion Jasnin, Tim Laugks, Miroslava Schaffer, and Margarete Schüler for help with cryoFIB in Martinsried. This work was supported by a Wellcome Trust Senior Research Fellowship to K.G. (090895/Z/09/Z), German Research Council grants DFG GR1990/2 to K.G. and DFG Me 854/12-1 to T.C.M., a Leverhulme Trust grant to K.G. (RPG-2012-519), a Wellcome Trust JIF award (060208/Z/00/Z) and a Wellcome Trust equipment grant (093305/Z/10/Z) to the Oxford Particle Imaging Centre, the Wellcome Trust core award 090532/Z/09/Z to the Wellcome Trust Centre for Human Genetics, the Wellcome Trust Strategic Award 091911 supporting advanced microscopy at Micron Oxford, a Wellcome Trust PhD studentship to M.G., a São Paulo Research Foundation Fellowship to J.C. (Fapesp 2013/24972-1), and a DFG Fellowship BO 4158/1-1 to J.B.B. We acknowledge the Helmholtz-Zentrum Berlin electron storage ring BESSY II for provision of synchrotron radiation at beamline U41-FSGM. The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under BioStruct-X (Grant agreement nos. 283570 and 226716).
Publisher Copyright:
© 2015 The Authors.
PY - 2015/12/17
Y1 - 2015/12/17
N2 - 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.
AB - 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.
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U2 - 10.1016/j.cell.2015.11.029
DO - 10.1016/j.cell.2015.11.029
M3 - Article
C2 - 26687357
AN - SCOPUS:84950262686
SN - 0092-8674
VL - 163
SP - 1692
EP - 1701
JO - Cell
JF - Cell
IS - 7
ER -