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Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends
The microtubule binding protein EB1 specifically targets the growing ends of microtubules in cells, where EB1 facilitates the interactions of cellular proteins with microtubule plus-ends. Microtubule end targeting of EB1 has been attributed to high-affinity binding of EB1 to GTP-tubulin that is pres...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6742484/ https://www.ncbi.nlm.nih.gov/pubmed/31478831 http://dx.doi.org/10.7554/eLife.48117 |
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author | Reid, Taylor A Coombes, Courtney Mukherjee, Soumya Goldblum, Rebecca R White, Kyle Parmar, Sneha McClellan, Mark Zanic, Marija Courtemanche, Naomi Gardner, Melissa K |
author_facet | Reid, Taylor A Coombes, Courtney Mukherjee, Soumya Goldblum, Rebecca R White, Kyle Parmar, Sneha McClellan, Mark Zanic, Marija Courtemanche, Naomi Gardner, Melissa K |
author_sort | Reid, Taylor A |
collection | PubMed |
description | The microtubule binding protein EB1 specifically targets the growing ends of microtubules in cells, where EB1 facilitates the interactions of cellular proteins with microtubule plus-ends. Microtubule end targeting of EB1 has been attributed to high-affinity binding of EB1 to GTP-tubulin that is present at growing microtubule ends. However, our 3D single-molecule diffusion simulations predicted a ~ 6000% increase in EB1 arrivals to open, tapered microtubule tip structures relative to closed lattice conformations. Using quantitative fluorescence, single-molecule, and electron microscopy experiments, we found that the binding of EB1 onto opened, structurally disrupted microtubules was dramatically increased relative to closed, intact microtubules, regardless of hydrolysis state. Correspondingly, in cells, the blunting of growing microtubule plus-ends by Vinblastine was correlated with reduced EB1 targeting. Together, our results suggest that microtubule structural recognition, based on a fundamental diffusion-limited binding model, facilitates the tip tracking of EB1 at growing microtubule ends. |
format | Online Article Text |
id | pubmed-6742484 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-67424842019-09-13 Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends Reid, Taylor A Coombes, Courtney Mukherjee, Soumya Goldblum, Rebecca R White, Kyle Parmar, Sneha McClellan, Mark Zanic, Marija Courtemanche, Naomi Gardner, Melissa K eLife Cell Biology The microtubule binding protein EB1 specifically targets the growing ends of microtubules in cells, where EB1 facilitates the interactions of cellular proteins with microtubule plus-ends. Microtubule end targeting of EB1 has been attributed to high-affinity binding of EB1 to GTP-tubulin that is present at growing microtubule ends. However, our 3D single-molecule diffusion simulations predicted a ~ 6000% increase in EB1 arrivals to open, tapered microtubule tip structures relative to closed lattice conformations. Using quantitative fluorescence, single-molecule, and electron microscopy experiments, we found that the binding of EB1 onto opened, structurally disrupted microtubules was dramatically increased relative to closed, intact microtubules, regardless of hydrolysis state. Correspondingly, in cells, the blunting of growing microtubule plus-ends by Vinblastine was correlated with reduced EB1 targeting. Together, our results suggest that microtubule structural recognition, based on a fundamental diffusion-limited binding model, facilitates the tip tracking of EB1 at growing microtubule ends. eLife Sciences Publications, Ltd 2019-09-03 /pmc/articles/PMC6742484/ /pubmed/31478831 http://dx.doi.org/10.7554/eLife.48117 Text en © 2019, Reid et al http://creativecommons.org/licenses/by/4.0/ 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 | Cell Biology Reid, Taylor A Coombes, Courtney Mukherjee, Soumya Goldblum, Rebecca R White, Kyle Parmar, Sneha McClellan, Mark Zanic, Marija Courtemanche, Naomi Gardner, Melissa K Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title | Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title_full | Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title_fullStr | Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title_full_unstemmed | Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title_short | Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends |
title_sort | structural state recognition facilitates tip tracking of eb1 at growing microtubule ends |
topic | Cell Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6742484/ https://www.ncbi.nlm.nih.gov/pubmed/31478831 http://dx.doi.org/10.7554/eLife.48117 |
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