Project Details
Projekt
Dissecting the biogenesis of Vaccinia Virus membranes using cryo-electron tomography
Subject Area
Virology
Term
from 2005 to 2014
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 5455996
Vaccinia virus (VV), a member of the poxviridae a family of large DNA viruses, was used successfully to eradicate variola virus, the cause of smallpox. Currently, VV vaccination is the only effective means to prevent deliberate smallpox dissemination. VV recombinants are also used as viral vectors to induce immunity to other pathogens or to treat certain forms of cancer. The structure and assembly of VV, the prototype member of the poxvirus family, is only partially understood and controversial. We will use cryo-electron tomography (cryo-ET) on whole cells to study the defined VV disassembly intermediates as well as VV as it leaves the cell, early and late in infection, respectively. These viral particles are located at the edge of infected cells and will be analyzed in 3D by cryo-ET using rapidly (plunge-) frozen cells grown on EM grids. VV-assembly, known to occur close to the nucleus in thicker parts of the cell, will be studied using frozen hydrated sections of high pressure frozen infected cells, in a close collaboration with the group of Dubochet (Lausanne). We will focus on imaging and reconstructing by cryo-ET the viral precursor membranes, that ultimately make up the mature virion, in particular their relation to and connections with cellular membranes. Together with a recent cryo-ET study on intact, purified VV particles, carried out by the coinvestigator, these studies should enable us to make substantial progress in understanding the structure and assembly of VV.
DFG Programme
Priority Programmes
