Structural Model for Tubulin Recognition and Deformation by Kinesin-13 Microtubule Depolymerases

SummaryTo elucidate the structural basis of the mechanism of microtubule depolymerization by kinesin-13s, we analyzed complexes of tubulin and the Drosophila melanogaster kinesin-13 KLP10A by electron microscopy (EM) and fluorescence polarization microscopy. We report a nanometer-resolution (1.1 nm) cryo-EM three-dimensional structure of the KLP10A head domain (KLP10AHD) bound to curved tubulin. We found that binding of KLP10AHD induces a distinct tubulin configuration with displacement (shear) between tubulin subunits in addition to curvature. In this configuration, the kinesin-binding site differs from that in straight tubulin, providing an explanation for the distinct interaction modes of kinesin-13s with the microtubule lattice or its ends. The KLP10AHD-tubulin interface comprises three areas of interaction, suggesting a crossbow-type tubulin-bending mechanism. These areas include the kinesin-13 family conserved KVD residues, and as predicted from the crossbow model, mutating these residues changes the orientation and mobility of KLP10AHDs interacting with the microtubule.

Graphical AbstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► Kinesin-13 tubulin complex structure reported at nanometer resolution ► Inter- and intradimer tubulin heterodimer interfaces are unambiguously identified ► A distinct curved-sheared tubulin conformation is described ► Insights provided into how kinesin-13s recognize and depolymerize microtubule ends