Nogales, E, Ramey, VH and Wang, HW. Cryo-EM studies of structural intermediates in microtubule dynamic instability and interaction of microtubules with kinetochore complexes. METHODS CELL BIOLOGY 95:129-156, 2010

Abstract The existence of structural intermediates in the processes of microtubule assembly and disassembly, and their relationship with the nucleotide state of tubulin, have been the subject of significant study and recent controversy. The first part of this chapter describes experiments and methods designed to characterize, using cryo-electron microscopy (cryo-EM) and image analysis, the structure of stabilized tubulin assemblies that we propose mimic the growth and shortening states at microtubule ends. We further put forward the idea that these intermediates have important biological functions, especially during cellular processes where the dynamic character of microtubules is essential. One such process is the attachment of spindle microtubules to kinetochores in eukaryotic cell divi......