Structural state recognition facilitates tip tracking of eb1 at growing microtubule ends

Taylor A. Reid, Courtney Coombes, Soumya Mukherjee, Rebecca R. Goldblum, Kyle White, Sneha Parmar, Mark McClellan, Marija Zanic, Naomi Courtemanche, Melissa K. Gardner

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

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.

Original languageEnglish (US)
Article numbere48117
JournaleLife
Volume8
DOIs
StatePublished - Sep 2019

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health grant NIGMS R01-GM103833 and R35-GM126974 and National Science Foundation CAREER award 1350741 to MKG. MZ acknowledges the support of National Institutes of Health grant R35-GM119552, the Human Frontier Science Program and the Searle Scholars Program. The authors have no conflicts of interest to disclose. Parts of this work were carried out in the Characterization Facility, University of Minnesota, a member of the NSF-funded Materials Research Facilities Network (www.mrfn.org) via the MRSEC program. We thank Drs. Joe Howard and Ron Vale for gifting reagents used as part of this project. We thank Holly Goodson and the Gardner Laboratory for helpful discussions.

Publisher Copyright:
© Reid et al.

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