Engineering the Processive Run Length of the Kinesin Motor

@article{citeulike:1479768, abstract = {Conventional kinesin is a highly processive molecular motor that takes several hundred steps per encounter with a microtubule. Processive motility is believed to result from the coordinated, hand-over-hand motion of the two heads of the kinesin dimer, but the specific factors that determine kinesin's run length (distance traveled per microtubule encounter) are not known. Here, we show that the neck coiled-coil, a structure adjacent to the motor domain, plays an important role in governing the run length. By adding positive charge to the neck coiled-coil, we have created ultra-processive kinesin mutants that have fourfold longer run lengths than the wild-type motor, but that have normal ATPase activity and motor velocity. Conversely, adding negative charge on the neck coiled-coil decreases the run length. The gain in processivity can be suppressed by either proteolytic cleavage of tubulin's negatively charged COOH terminus or by high salt concentrations. Therefore, modulation of processivity by the neck coiled-coil appears to involve an electrostatic tethering interaction with the COOH terminus of tubulin. The ability to readily increase kinesin processivity by mutation, taken together with the strong sequence conservation of the neck coiled-coil, suggests that evolutionary pressures may limit kinesin's run length to optimize its in vivo function. 10.1083/jcb.151.5.1093}, author = {Thorn, Kurt S. and Ubersax, Jeffrey A. and Vale, Ronald D. }, citeulike-article-id = {1479768}, comment = {e10}, doi = {10.1083/jcb.151.5.1093}, journal = {J. Cell Biol.}, keywords = {charge, electrostatics, experiment, kinesin, motor, processivity}, month = {November}, number = {5}, pages = {1093--1100}, posted-at = {2007-07-25 03:42:05}, priority = {0}, title = {Engineering the Processive Run Length of the Kinesin Motor}, url = {http://dx.doi.org/10.1083/jcb.151.5.1093}, volume = {151}, year = {2000} }

TY - JOUR ID - citeulike:1479768 L3 - citeulike-article-id:1479768 TI - Engineering the Processive Run Length of the Kinesin Motor JF - J. Cell Biol. VL - 151 IS - 5 SP - 1093 EP - 1100 N2 - Conventional kinesin is a highly processive molecular motor that takes several hundred steps per encounter with a microtubule. Processive motility is believed to result from the coordinated, hand-over-hand motion of the two heads of the kinesin dimer, but the specific factors that determine kinesin's run length (distance traveled per microtubule encounter) are not known. Here, we show that the neck coiled-coil, a structure adjacent to the motor domain, plays an important role in governing the run length. By adding positive charge to the neck coiled-coil, we have created ultra-processive kinesin mutants that have fourfold longer run lengths than the wild-type motor, but that have normal ATPase activity and motor velocity. Conversely, adding negative charge on the neck coiled-coil decreases the run length. The gain in processivity can be suppressed by either proteolytic cleavage of tubulin's negatively charged COOH terminus or by high salt concentrations. Therefore, modulation of processivity by the neck coiled-coil appears to involve an electrostatic tethering interaction with the COOH terminus of tubulin. The ability to readily increase kinesin processivity by mutation, taken together with the strong sequence conservation of the neck coiled-coil, suggests that evolutionary pressures may limit kinesin's run length to optimize its in vivo function. 10.1083/jcb.151.5.1093 KW - charge KW - electrostatics KW - experiment KW - kinesin KW - motor KW - processivity N1 - e10 AU - Thorn, Kurt AU - Ubersax, Jeffrey AU - Vale, Ronald PY - 2000/11/20/ UR - http://dx.doi.org/10.1083/jcb.151.5.1093 ER -