Project Details
Projekt
Structural analysis of herpesvirus assembly by cryo electron tomography
Applicant
Professor Dr. Kay Grünewald
Subject Area
Virology
Term
from 2005 to 2012
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 13491680
Applying electron cryo tomography to virus infected cells we aim to elucidate the involvement and interaction of macromolecular complexes from both, the virus and the host cell, in situ in a near native state. Collaborating with the groups of Beate Sodeik (Hannover) and Hans-Georg Kräusslich (Heidelberg) we will investigate aspects of herpesvirus assembly and human immunodeficiency virus budding at the plasma membrane, respectively. The project is based on our earlier studies on the overall architecture of isolated virus particles for both viruses and targets on extending the technique to virus infected host cells. Preliminary data obtained from uninfected cells show that peripheral cell regions can be sufficiently penetrated by the electron beam to allow acquisition of tomographic data sets under low dose conditions. Additionally, we will establish the sectioning of host cells in the frozen hydrated state to overcome the inherent limitations for electron cryo-tomography imposed by the thickness of the cellular specimens. Applying a hybrid approach of fluorescence microscopy and electron cryo-microscopy we will be able to focus on defined states of the assembly process. The chosen technique allows linking existing results from biochemical studies to native structural information at the sub-cellular level.
DFG Programme
Priority Programmes
