Cryo-electron tomography of filament assemblies and their organizers

Summary:

Cryo-electron tomography (cryo-ET) combined with subtomogram averaging allows structure determination in complex environments, such as cells, often recalcitrant to other structural biology approaches. While the achievable resolutions in most cases remain lower than possible with single particle analysis cryo-electron microscopy, the contextual insight provided by the tomography approach delivers a unique flavor of insights. Such insights can be crucial for understanding the architecture of filament assemblies (e.g., the actin-rich protrusions of migrating cells) and the role of their organizers. To make the best use of the structural and contextual information contained in tomograms, we assembled workflows and software pipelines that allow for rapid ultrastructural analysis of parallel and meshwork-like filament assemblies, as well as structure determination of proteins organizing such macromolecular constructions.
During this presentation, I will give a short overview of the pipelines and showcase their application, mainly in the context of actin filament networks.