Structural Studies of Viral Pathogens Using Cryo-EM and 3D Reconstruction
These researchers use cryo-electron microscopy (cryo-EM) and coordinated three-dimensional reconstruction techniques to study the macromolecular structures of bacterial phages, enveloped Alphaviruses and retroviruses. They are working on three projects that use MSI:
- Bacteriophage Phi29: 3D reconstruction of Phi29 mutants to 1-2 nanometer resolution: The process involves employing a number of graphical and parallel computational software packages to isolate images of individual virus particles, characterize the parameters involved in microscopy, determine the orientation and origin of each virus particle, compute a reconstruction map, and dock the known atomic structures of the protein components into the reconstruction map.
- Structural studies of alphavirus membrane fusion in situ: Enveloped Sindbis virus infects host cells through membrane fusion of viral and cellular membranes, a process directed by the viral fusion protein E1 at low pH. In order to understand the molecular mechanism that underlines membrane fusion, virus-liposome complexes were produced at different pH conditions. Analysis and comparison of the reconstruction maps will lead to understanding the protein conformational and oligomerizational changes during such membrane remodeling process.
- Structural studies of Human Immunodeficiency Virus type 2 and Human T-cell Leukemia virus type 1: Cryo-electron tomography reconstruction mapping will be used to illustrate the surface protein organization, membrane envelope, and the internal protein lattices. This structural information will provide insights into the molecular mechanism of assembly process of the virus.
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