Visualizing cellular processes at the molecular level by cryo-electron tomography.
The cellular landscape rapidly changes throughout the biological processes that transpire within a cell. For example, the cytoskeleton is remodeled within fractions of a second. Therefore, reliable structural analysis of the cell requires approaches that allow for instantaneous arrest of functional states of a given process while offering the best possible preservation of the delicate cellular structure. Electron tomography of vitrified but otherwise unaltered cells (cryo-ET) has proven to be the method of choice for three-dimensional (3D) reconstruction of cellular architecture at a resolution of 4-6 nm. Through the use of cryo-ET, the 3D organization of macromolecular complexes and organelles can be studied in their native environment in the cell. In this Commentary, we focus on the application of cryo-ET to study eukaryotic cells - in particular, the cytoskeletal-driven processes that are involved in cell movements, filopodia protrusion and viral entry. Finally, we demonstrate the potential of cryo-ET to determine structures of macromolecular complexes in situ, such as the nuclear pore complex.
Ben-Harush
Kfir
K
Maimon
Tal
T
Patla
Israel
I
Villa
Elizabeth
E
Medalia
Ohad
O
D000818:Animals; D002468:Cell Physiological Phenomena; D020285:Cryoelectron Microscopy / Q000295:instrumentation / Q000379:methods*; D003599:Cytoskeleton / Q000648:ultrastructure; D055032:Electron Microscope Tomography / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D022022:Nuclear Pore / Q000648:ultrastructure