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Template-free 13-protofilament microtubule-MAP assembly visualized at 8 A resolution.


ABSTRACT: 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.

SUBMITTER: Fourniol FJ 

PROVIDER: S-EPMC3003314 | biostudies-literature | 2010 Nov

REPOSITORIES: biostudies-literature

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Template-free 13-protofilament microtubule-MAP assembly visualized at 8 A resolution.

Fourniol Franck J FJ   Sindelar Charles V CV   Amigues Béatrice B   Clare Daniel K DK   Thomas Geraint G   Perderiset Mylène M   Francis Fiona F   Houdusse Anne A   Moores Carolyn A CA  

The Journal of cell biology 20101025 3


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 int  ...[more]

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