Structural basis of microtubule destabilization by GTP hydrolysis

Abstract

Microtubules are cytoskeletal filaments that dynamically grow and shrink to support vital biological functions. Their behavior is regulated by GTP hydrolysis in tubulin incorporated into the lattice, but the underlying chemical transitions and their impact on microtubule stability remain unclear. Using cryo-electron microscopy, we resolved microtubule structures at 1.9-2.2 [A] in four different nucleotide states, revealing the roles of amino acids, ions, and hundreds of water molecules in the hydrolysis process. We found that compaction of the GTP-bound lattice into a more stable pre-hydrolysis state activates a catalytic water molecule for {gamma}-phosphate cleavage. Hydrolysis intermediates are stabilized by strengthened lattice contacts, but in the final GDP state, release of reaction products disrupts these contacts and destabilizes the lattice. Our results explain how chemical changes in tubulins nucleotide site drive lattice transitions and offer an atomistic framework for understanding microtubule dynamic instability.