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

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. B...

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Autores principales: Driver, Jonathan W, Geyer, Elisabeth A, Bailey, Megan E, Rice, Luke M, Asbury, Charles L
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5515574/
https://www.ncbi.nlm.nih.gov/pubmed/28628007
http://dx.doi.org/10.7554/eLife.28433
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author Driver, Jonathan W
Geyer, Elisabeth A
Bailey, Megan E
Rice, Luke M
Asbury, Charles L
author_facet Driver, Jonathan W
Geyer, Elisabeth A
Bailey, Megan E
Rice, Luke M
Asbury, Charles L
author_sort Driver, Jonathan W
collection PubMed
description 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 spring-like 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 β-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. DOI: http://dx.doi.org/10.7554/eLife.28433.001
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spelling pubmed-55155742017-07-19 Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips Driver, Jonathan W Geyer, Elisabeth A Bailey, Megan E Rice, Luke M Asbury, Charles L eLife Biophysics and Structural Biology 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 spring-like 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 β-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. DOI: http://dx.doi.org/10.7554/eLife.28433.001 eLife Sciences Publications, Ltd 2017-06-19 /pmc/articles/PMC5515574/ /pubmed/28628007 http://dx.doi.org/10.7554/eLife.28433 Text en © 2017, Driver et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biophysics and Structural Biology
Driver, Jonathan W
Geyer, Elisabeth A
Bailey, Megan E
Rice, Luke M
Asbury, Charles L
Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title_full Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title_fullStr Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title_full_unstemmed Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title_short Direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
title_sort direct measurement of conformational strain energy in protofilaments curling outward from disassembling microtubule tips
topic Biophysics and Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5515574/
https://www.ncbi.nlm.nih.gov/pubmed/28628007
http://dx.doi.org/10.7554/eLife.28433
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