ausblenden:
Schlagwörter:
Cryo-electron tomography workflows
In -cell structural analysis
Nanoscale actin architecture
Subtomogram averaging
Actin polarity
Force generation
molecular sociology
electron tomograms
structural biology
eukaryotic
cells
architecture
complex
organization
filaments
platform
toolbox
Cell Biology
Zusammenfassung:
Actin contributes to an exceptionally wide range of cellular processes through the assembly and disassembly of highly dynamic and ordered structures. Visualizing these structures in cells can help us understand how the molecular players of the actin machinery work together to produce force-generating systems. In recent years, cryo-electron tomography (cryo-ET) has become the method of choice for structural analysis of the cell interior at the molecular scale. Here we review advances in cryo-ET workflows that have enabled this transformation, especially the automation of sample preparation procedures, data collection, and processing. We discuss new structural analyses of dynamic actin assemblies in cryo-preserved cells, which have provided mechanistic in-sights into actin assembly and function at the nanoscale. Finally, we highlight the latest visual proteomics studies of actin filaments and their interactors reaching sub-nanometer resolutions in cells.
