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First published online December 5, 2008; 10.1105/tpc.108.063362

The Plant Cell 20:3346-3358 (2008)
© 2008 American Society of Plant Biologists

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The C-Terminal Variable Region Specifies the Dynamic Properties of Arabidopsis Microtubule-Associated Protein MAP65 Isotypes[C],[W]

Andrei P. Smertenkoa,1, Despina Kaloritia,1, Hsin-Yu Changa, Jindriska Fiserovaa,b, Zdenek Opatrnyb and Patrick J. Husseya,2

a Integrative Cell Biology Laboratory, School of Biological and Biomedical Sciences, University of Durham, Durham DH1 3LE, United Kingdom
b Department of Plant Physiology, Charles University, Faculty of Science, 128 44 Prague 2,Czech Republic

2 Address correspondence to p.j.hussey{at}durham.ac.uk.

The microtubule-associated protein, MAP65, is a member of a family of divergent microtubule-associated proteins from different organisms generally involved in maintaining the integrity of the central spindle in mitosis. The dicotyledon Arabidopsis thaliana and the monocotyledon rice (Oryza sativa) genomes contain 9 and 11 MAP65 genes, respectively. In this work, we show that the majority of these proteins fall into five phylogenetic clades, with the greatest variation between clades being in the C-terminal random coil domain. At least one Arabidopsis and one rice isotype is within each clade, indicating a functional specification for the C terminus. In At MAP65-1, the C-terminal domain is a microtubule binding region (MTB2) harboring the phosphorylation sites that control its activity. The At MAP65 isotypes show differential localization to microtubule arrays and promote microtubule polymerization with variable efficiency in a MTB2-dependent manner. In vivo studies demonstrate that the dynamics of the association and dissociation of different MAP65 isotypes with microtubules can vary up to 10-fold and that this correlates with their ability to promote microtubule polymerization. Our data demonstrate that the C-terminal variable region, MTB2, determines the dynamic properties of individual isotypes and suggest that slower turnover is conditional for more efficient microtubule polymerization.




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