TY - JOUR
T1 - Structural model for tubulin recognition and deformation by kinesin-13 microtubule depolymerases
AU - Asenjo, Ana B.
AU - Chatterjee, Chandrima
AU - Tan, Dongyan
AU - DePaoli, Vania
AU - Rice, William J.
AU - Diaz-Avalos, Ruben
AU - Silvestry, Mariena
AU - Sosa, Hernando
N1 - Funding Information:
We thank J. Hargitai and S. Wang for help with installing and running software in the high-performance computer cluster at the Albert Einstein College of Medicine (AECOM); F. Macaluso, L. Gunther, and members of the AECOM Analytical Imaging Facility for help with EM; and D. Sharp and G. Gerfen for discussions and critical reading of the manuscript. Part of this investigation was conducted in a facility constructed with support from the Research Facilities Improvement Program of the National Center for Research Resources, National Institutes of Health (NIH, grant C06 RR017528-01-CEM). The 300 keV microscope at the New York Structural Biology Center was purchased with funds from NIH grant S10 RR17291. This project was supported by NIH grant R01-GM083338 to H.S.
PY - 2013
Y1 - 2013
N2 - To 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. Kinesin-13 proteins are microtubule (MT) depolymerases that play a key role in modulating MT dynamics in a variety of cellular processes. How kinesin-13s induce depolymerization, rather than walking along MTs like most other kinesins, is not clear. Structural analysis by Sosa and colleagues shows that binding of the kinesin-13 catalytic domain to tubulin alters its conformation to one that is incompatible with the formation of MTs and instead favors binding of kinesin-13 over that of other kinesin proteins.
AB - To 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. Kinesin-13 proteins are microtubule (MT) depolymerases that play a key role in modulating MT dynamics in a variety of cellular processes. How kinesin-13s induce depolymerization, rather than walking along MTs like most other kinesins, is not clear. Structural analysis by Sosa and colleagues shows that binding of the kinesin-13 catalytic domain to tubulin alters its conformation to one that is incompatible with the formation of MTs and instead favors binding of kinesin-13 over that of other kinesin proteins.
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U2 - 10.1016/j.celrep.2013.01.030
DO - 10.1016/j.celrep.2013.01.030
M3 - Article
C2 - 23434508
AN - SCOPUS:84875806836
SN - 2211-1247
VL - 3
SP - 759
EP - 768
JO - Cell Reports
JF - Cell Reports
IS - 3
ER -