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Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution

Microtubule-associated proteins (MAPs) are essential for regulating and organizing cellular microtubules (MTs). However, our mechanistic understanding of MAP function is limited by a lack of detailed structural information. Using cryo-electron microscopy and single particle algorithms, we solved the...

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Autores principales: Fourniol, Franck J., Sindelar, Charles V., Amigues, Béatrice, Clare, Daniel K., Thomas, Geraint, Perderiset, Mylène, Francis, Fiona, Houdusse, Anne, Moores, Carolyn A.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3003314/
https://www.ncbi.nlm.nih.gov/pubmed/20974813
http://dx.doi.org/10.1083/jcb.201007081
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author Fourniol, Franck J.
Sindelar, Charles V.
Amigues, Béatrice
Clare, Daniel K.
Thomas, Geraint
Perderiset, Mylène
Francis, Fiona
Houdusse, Anne
Moores, Carolyn A.
author_facet Fourniol, Franck J.
Sindelar, Charles V.
Amigues, Béatrice
Clare, Daniel K.
Thomas, Geraint
Perderiset, Mylène
Francis, Fiona
Houdusse, Anne
Moores, Carolyn A.
author_sort Fourniol, Franck J.
collection PubMed
description Microtubule-associated proteins (MAPs) are essential for regulating and organizing cellular microtubules (MTs). However, our mechanistic understanding of MAP function is limited by a lack of detailed structural information. Using cryo-electron microscopy and single particle algorithms, we solved the 8 Å structure of doublecortin (DCX)-stabilized MTs. Because of DCX’s unusual ability to specifically nucleate and stabilize 13-protofilament MTs, our reconstruction provides unprecedented insight into the structure of MTs with an in vivo architecture, and in the absence of a stabilizing drug. DCX specifically recognizes the corner of four tubulin dimers, a binding mode ideally suited to stabilizing both lateral and longitudinal lattice contacts. A striking consequence of this is that DCX does not bind the MT seam. DCX binding on the MT surface indirectly stabilizes conserved tubulin–tubulin lateral contacts in the MT lumen, operating independently of the nucleotide bound to tubulin. DCX’s exquisite binding selectivity uncovers important insights into regulation of cellular MTs.
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spelling pubmed-30033142011-05-01 Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution Fourniol, Franck J. Sindelar, Charles V. Amigues, Béatrice Clare, Daniel K. Thomas, Geraint Perderiset, Mylène Francis, Fiona Houdusse, Anne Moores, Carolyn A. J Cell Biol Research Articles Microtubule-associated proteins (MAPs) are essential for regulating and organizing cellular microtubules (MTs). However, our mechanistic understanding of MAP function is limited by a lack of detailed structural information. Using cryo-electron microscopy and single particle algorithms, we solved the 8 Å structure of doublecortin (DCX)-stabilized MTs. Because of DCX’s unusual ability to specifically nucleate and stabilize 13-protofilament MTs, our reconstruction provides unprecedented insight into the structure of MTs with an in vivo architecture, and in the absence of a stabilizing drug. DCX specifically recognizes the corner of four tubulin dimers, a binding mode ideally suited to stabilizing both lateral and longitudinal lattice contacts. A striking consequence of this is that DCX does not bind the MT seam. DCX binding on the MT surface indirectly stabilizes conserved tubulin–tubulin lateral contacts in the MT lumen, operating independently of the nucleotide bound to tubulin. DCX’s exquisite binding selectivity uncovers important insights into regulation of cellular MTs. The Rockefeller University Press 2010-11-01 /pmc/articles/PMC3003314/ /pubmed/20974813 http://dx.doi.org/10.1083/jcb.201007081 Text en © 2010 Fourniol 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
Fourniol, Franck J.
Sindelar, Charles V.
Amigues, Béatrice
Clare, Daniel K.
Thomas, Geraint
Perderiset, Mylène
Francis, Fiona
Houdusse, Anne
Moores, Carolyn A.
Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title_full Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title_fullStr Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title_full_unstemmed Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title_short Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution
title_sort template-free 13-protofilament microtubule–map assembly visualized at 8 å resolution
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3003314/
https://www.ncbi.nlm.nih.gov/pubmed/20974813
http://dx.doi.org/10.1083/jcb.201007081
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