Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends in cells
Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends in cells
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 conversion of growing microtubule ends from a tapered into a blunt configuration resulted in 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.
Coombes Courtney、Parmar Sneha、Mukherjee Soumya、White Kyle、Zanic Marija、McClellan Mark、Courtemanche Naomi、Goldblum Rebecca R.、Gardner Melissa K.、Reid Taylor A.
Department of Genetics, Cell Biology, and Development, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of MinnesotaDepartments of Cell and Developmental Biology, Chemical and Biomolecular Engineering, and Biochemistry, Vanderbilt UniversityDepartment of Genetics, Cell Biology, and Development, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of MinnesotaMedical Scientist Training Program and Department of Biochemistry, Molecular Biology, and Biophysics, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of MinnesotaDepartment of Genetics, Cell Biology, and Development, University of Minnesota
细胞生物学生物物理学分子生物学
Coombes Courtney,Parmar Sneha,Mukherjee Soumya,White Kyle,Zanic Marija,McClellan Mark,Courtemanche Naomi,Goldblum Rebecca R.,Gardner Melissa K.,Reid Taylor A..Structural state recognition facilitates tip tracking of EB1 at growing microtubule ends in cells[EB/OL].(2025-03-28)[2025-05-05].https://www.biorxiv.org/content/10.1101/636092.点此复制
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