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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...

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Autores principales: Reid, Taylor A, Coombes, Courtney, Mukherjee, Soumya, Goldblum, Rebecca R, White, Kyle, Parmar, Sneha, McClellan, Mark, Zanic, Marija, Courtemanche, Naomi, Gardner, Melissa K
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
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.
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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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