Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips

被引:34
|
作者
Driver, Jonathan W. [1 ]
Geyer, Elisabeth A. [2 ,3 ]
Bailey, Megan E. [1 ]
Rice, Luke M. [2 ,3 ]
Asbury, Charles L. [1 ]
机构
[1] Univ Washington, Dept Physiol & Biophys, Seattle, WA 98195 USA
[2] UT Southwestern Med Ctr Dallas, Dept Biophys, Dallas, TX 75390 USA
[3] UT Southwestern Med Ctr Dallas, Dept Biochem, Dallas, TX 75390 USA
来源
ELIFE | 2017年 / 6卷
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
FORCE PRODUCTION; DYNAMICS; SPINDLE; DEPOLYMERIZATION; KINETOCHORES; MECHANISM; MOVEMENT; RIGIDITY; KINESIN; MITOSIS;
D O I
10.7554/eLife.28433
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Disassembling microtubules can generate movement independently of motor enzymes, especially at kinetochores where they drive chromosome motility. A popular explanation is the 'conformational wave' model, in which protofilaments pull on the kinetochore as they curl outward from a disassembling tip. But whether protofilaments can work efficiently via this springlike mechanism has been unclear. By modifying a previous assay to use recombinant tubulin and feedback-controlled laser trapping, we directly demonstrate the spring-like elasticity of curling protofilaments. Measuring their mechanical work output suggests they carry 25% of the energy of GTP hydrolysis as bending strain, enabling them to drive movement with efficiency similar to conventional motors. Surprisingly, a p-tubulin mutant that dramatically slows disassembly has no effect on work output, indicating an uncoupling of disassembly speed from protofilament strain. These results show the wave mechanism can make a major contribution to kinetochore motility and establish a direct approach for measuring tubulin mechano-chemistry.
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页数:18
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