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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Rockefeller University Press
2012
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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. |
format | Online Article Text |
id | pubmed-3413364 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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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