Abstract
Cryo-electron microscopy (Cryo-EM) has become a major tool in the structural characterization of large macromolecular assemblies, their architecture, interactions with different ligands, and the regulation of their function. I will present two different examples of how cryo-EM is being used in my lab to understand the molecular mechanisms of complex biological systems. During division the eukaryotic cell needs to accurately segregate its genetic material between daughter cells. This process involves the interaction of the microtubule mitotic spindle with special regions on chromosomes called kinetochores. Errors, which result in misplaced chromosomes, can lead to cancer or death. We have visualized the interaction of microtubules with two kinetochore components, the yeast Dam1 and the human Ndc80 complexes, using cryo-electron microscopy and image reconstruction. Interestingly, both complexes oligomerize on the surface of the microtubule, a property that is essential for their capacity for harness the energy of microtubule depolymerization for chromosome movement.
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