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Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy

Microtubules are dynamic polymers that stochastically switch between growing and shrinking phases. Microtubule dynamics are regulated by guanosine triphosphate (GTP) hydrolysis by β-tubulin, but the mechanism of this regulation remains elusive because high-resolution microtubule structures have only...

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Autores principales: Yajima, Hiroaki, Ogura, Toshihiko, Nitta, Ryo, Okada, Yasushi, Sato, Chikara, Hirokawa, Nobutaka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3413364/
https://www.ncbi.nlm.nih.gov/pubmed/22851320
http://dx.doi.org/10.1083/jcb.201201161
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author Yajima, Hiroaki
Ogura, Toshihiko
Nitta, Ryo
Okada, Yasushi
Sato, Chikara
Hirokawa, Nobutaka
author_facet Yajima, Hiroaki
Ogura, Toshihiko
Nitta, Ryo
Okada, Yasushi
Sato, Chikara
Hirokawa, Nobutaka
author_sort Yajima, Hiroaki
collection PubMed
description Microtubules are dynamic polymers that stochastically switch between growing and shrinking phases. Microtubule dynamics are regulated by guanosine triphosphate (GTP) hydrolysis by β-tubulin, but the mechanism of this regulation remains elusive because high-resolution microtubule structures have only been revealed for the guanosine diphosphate (GDP) state. In this paper, we solved the cryoelectron microscopy (cryo-EM) structure of microtubule stabilized with a GTP analogue, guanylyl 5′-α,β-methylenediphosphonate (GMPCPP), at 8.8-Å resolution by developing a novel cryo-EM image reconstruction algorithm. In contrast to the crystal structures of GTP-bound tubulin relatives such as γ-tubulin and bacterial tubulins, significant changes were detected between GMPCPP and GDP-taxol microtubules at the contacts between tubulins both along the protofilament and between neighboring protofilaments, contributing to the stability of the microtubule. These findings are consistent with the structural plasticity or lattice model and suggest the structural basis not only for the regulatory mechanism of microtubule dynamics but also for the recognition of the nucleotide state of the microtubule by several microtubule-binding proteins, such as EB1 or kinesin.
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spelling pubmed-34133642013-02-06 Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy Yajima, Hiroaki Ogura, Toshihiko Nitta, Ryo Okada, Yasushi Sato, Chikara Hirokawa, Nobutaka J Cell Biol Research Articles Microtubules are dynamic polymers that stochastically switch between growing and shrinking phases. Microtubule dynamics are regulated by guanosine triphosphate (GTP) hydrolysis by β-tubulin, but the mechanism of this regulation remains elusive because high-resolution microtubule structures have only been revealed for the guanosine diphosphate (GDP) state. In this paper, we solved the cryoelectron microscopy (cryo-EM) structure of microtubule stabilized with a GTP analogue, guanylyl 5′-α,β-methylenediphosphonate (GMPCPP), at 8.8-Å resolution by developing a novel cryo-EM image reconstruction algorithm. In contrast to the crystal structures of GTP-bound tubulin relatives such as γ-tubulin and bacterial tubulins, significant changes were detected between GMPCPP and GDP-taxol microtubules at the contacts between tubulins both along the protofilament and between neighboring protofilaments, contributing to the stability of the microtubule. These findings are consistent with the structural plasticity or lattice model and suggest the structural basis not only for the regulatory mechanism of microtubule dynamics but also for the recognition of the nucleotide state of the microtubule by several microtubule-binding proteins, such as EB1 or kinesin. The Rockefeller University Press 2012-08-06 /pmc/articles/PMC3413364/ /pubmed/22851320 http://dx.doi.org/10.1083/jcb.201201161 Text en © 2012 Yajima et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Articles
Yajima, Hiroaki
Ogura, Toshihiko
Nitta, Ryo
Okada, Yasushi
Sato, Chikara
Hirokawa, Nobutaka
Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title_full Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title_fullStr Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title_full_unstemmed Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title_short Conformational changes in tubulin in GMPCPP and GDP-taxol microtubules observed by cryoelectron microscopy
title_sort conformational changes in tubulin in gmpcpp and gdp-taxol microtubules observed by cryoelectron microscopy
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3413364/
https://www.ncbi.nlm.nih.gov/pubmed/22851320
http://dx.doi.org/10.1083/jcb.201201161
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