In vivo dynamics and differential microtubule-binding activities of MAP65 proteins

Daniël Van Damme, Kris Van Poucke, Emmanuel Boutant, Christophe Ritzenthaler, Dirk Inzé, Danny Geelen

    Research output: Contribution to journalA2: International peer reviewed article (not A1-type)peer-review

    Abstract

    Plant cells produce different microtubule arrays that are essential for cell division and morphogenesis without equivalent in other eukaryotes. Microtubule-associated proteins influence the behavior of microtubules that is presumed to culminate into transitions from one array to another. We analyzed the microtubule-binding properties of three Arabidopsis (Arabidopsis thaliana) members, AtMAP65-1, AtMAP65-4, and AtMAP65-5, in live cells using laser scanning confocal microscopy. Depending on the overall organization of the cortical array, AtMAP65-1-GFP (green fluorescent protein) and AtMAP65-5-GFP associated with a subset of microtubules. In cells containing both coaligned and oblique microtubules, AtMAP65-1-GFP and AtMAP65-5-GFP tended to be associated with the coaligned microtubules. Cortical microtubules labeled with AtMAP65-1-GFP and AtMAP65-5-GFP appeared as thick bundles and showed more resistance to microtubule-destabilizing drugs. The polymerization rates of AtMAP65-1-GFP and AtMAP65-5-GFP microtubules were similar to those of tubulin-GFP marked microtubules but were different from AtEB1a-GFP, a microtubule plus-end-binding EB1-like protein that stimulated polymerization. By contrast, depolymerization rates of AtMAP65-1-GFP- and AtMAP65-5-GFP-labeled microtubules were reduced. AtMAP65-1-GFP associated with polymerizing microtubules within a bundle, and with fixed microtubule termini, suggesting that AtMAP65-1's function is to bundle and stabilize adjacent microtubules of the cortex. Polymerization within a bundle took place in either direction so that bundling occurred between parallel or antiparallel aligned microtubules. AtMAP65-4-GFP did not label cortical microtubules or the preprophase band, despite continuous expression driven by the 35S promoter, and its subcellular localization was restricted to microtubules that rearranged to form a spindle and the polar sides of the spindle proper. The expression of AtMAP65-4 peaked at mitosis, in agreement with a function related to spindle formation, whereas AtMAP65-1 and AtMAP65-5 were expressed throughout the cell cycle.

    Original languageEnglish
    JournalPlant Physiology
    Volume136
    Issue number4
    Pages (from-to)3956-67
    Number of pages12
    ISSN0032-0889
    DOIs
    Publication statusPublished - 2004

    Keywords

    • Arabidopsis
    • Arabidopsis Proteins
    • Benzamides
    • Gene Expression Regulation, Plant
    • Green Fluorescent Proteins
    • Microscopy, Confocal
    • Microtubule-Associated Proteins
    • Microtubules
    • Mitosis
    • Nitrobenzenes
    • Organothiophosphorus Compounds
    • Plants, Genetically Modified
    • Protein Binding
    • Tobacco
    • Transcriptional Activation

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