The lattice as allosteric effector: Structural studies of alphabeta- and gamma-tubulin clarify the role of GTP in microtubule assembly

@article{citeulike:2744934, abstract = {GTP-dependent microtubule polymerization dynamics are required for cell division and are accompanied by domain rearrangements in the polymerizing subunit, {alpha}-tubulin. Two opposing models describe the role of GTP and its relationship to conformational change in {alpha}-tubulin. The allosteric model posits that unpolymerized {alpha}-tubulin adopts a more polymerization-competent conformation upon GTP binding. The lattice model posits that conformational changes occur only upon recruitment into the growing lattice. Published data support a lattice model, but are largely indirect and so the allosteric model has prevailed. We present two independent solution probes of the conformation of {alpha}-tubulin, the 2.3 A crystal structure of {gamma}-tubulin bound to GDP, and kinetic simulations to interpret the functional consequences of the structural data. These results (with our previous {gamma}-tubulin:GTP{gamma}S structure) support the lattice model by demonstrating that major domain rearrangements do not occur in eukaryotic tubulins in response to GTP binding, and that the unpolymerized conformation of {alpha}-tubulin differs significantly from the polymerized one. Thus, geometric constraints of lateral self-assembly must drive {alpha}-tubulin conformational changes, whereas GTP plays a secondary role to tune the strength of longitudinal contacts within the microtubule lattice. {alpha}-Tubulin behaves like a bent spring, resisting straightening until forced to do so by GTP-mediated interactions with the growing microtubule. Kinetic simulations demonstrate that resistance to straightening opposes microtubule initiation by specifically destabilizing early assembly intermediates that are especially sensitive to the strength of lateral interactions. These data provide new insights into the molecular origins of dynamic microtubule behavior. 10.1073/pnas.0801155105}, author = {Rice, Luke M. and Montabana, Elizabeth A. and Agard, David A. }, citeulike-article-id = {2744934}, doi = {10.1073/pnas.0801155105}, journal = {Proceedings of the National Academy of Sciences}, keywords = {allostery, kinetics, polymerization, simulation, structural\_change, xray-crystal-structure}, month = {April}, number = {14}, pages = {5378--5383}, posted-at = {2008-05-02 11:25:14}, priority = {2}, title = {The lattice as allosteric effector: Structural studies of {alpha}{beta}- and {gamma}-tubulin clarify the role of GTP in microtubule assembly}, url = {http://dx.doi.org/10.1073/pnas.0801155105}, volume = {105}, year = {2008} }

TY - JOUR ID - citeulike:2744934 TI - The lattice as allosteric effector: Structural studies of {alpha}{beta}- and {gamma}-tubulin clarify the role of GTP in microtubule assembly JF - Proceedings of the National Academy of Sciences VL - 105 IS - 14 SP - 5378 EP - 5383 N2 - GTP-dependent microtubule polymerization dynamics are required for cell division and are accompanied by domain rearrangements in the polymerizing subunit, {alpha}-tubulin. Two opposing models describe the role of GTP and its relationship to conformational change in {alpha}-tubulin. The allosteric model posits that unpolymerized {alpha}-tubulin adopts a more polymerization-competent conformation upon GTP binding. The lattice model posits that conformational changes occur only upon recruitment into the growing lattice. Published data support a lattice model, but are largely indirect and so the allosteric model has prevailed. We present two independent solution probes of the conformation of {alpha}-tubulin, the 2.3 A crystal structure of {gamma}-tubulin bound to GDP, and kinetic simulations to interpret the functional consequences of the structural data. These results (with our previous {gamma}-tubulin:GTP{gamma}S structure) support the lattice model by demonstrating that major domain rearrangements do not occur in eukaryotic tubulins in response to GTP binding, and that the unpolymerized conformation of {alpha}-tubulin differs significantly from the polymerized one. Thus, geometric constraints of lateral self-assembly must drive {alpha}-tubulin conformational changes, whereas GTP plays a secondary role to tune the strength of longitudinal contacts within the microtubule lattice. {alpha}-Tubulin behaves like a bent spring, resisting straightening until forced to do so by GTP-mediated interactions with the growing microtubule. Kinetic simulations demonstrate that resistance to straightening opposes microtubule initiation by specifically destabilizing early assembly intermediates that are especially sensitive to the strength of lateral interactions. These data provide new insights into the molecular origins of dynamic microtubule behavior. 10.1073/pnas.0801155105 KW - allostery KW - kinetics KW - polymerization KW - simulation KW - structural_change KW - xray-crystal-structure AU - Rice, Luke AU - Montabana, Elizabeth AU - Agard, David PY - 2008/04/08/ UR - http://dx.doi.org/10.1073/pnas.0801155105 ER -