Spindle assembly checkpoint protein dynamics reveal conserved and unsuspected roles in plant cell division.

BACKGROUND: In eukaryotes, the spindle assembly checkpoint (SAC) ensures that chromosomes undergoing mitosis do not segregate until they are properly attached to the microtubules of the spindle. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the mechanism underlying this surveillance mechanism in plants, by characterising the orthogolous SAC proteins BUBR1, BUB3 and MAD2 from Arabidopsis. We showed that the cell cycle-regulated BUBR1, BUB3.1 and MAD2 proteins interacted physically with each other. Furthermore, BUBR1 and MAD2 interacted specifically at chromocenters. Following SAC activation by global defects in spindle assembly, these three interacting partners localised to unattached kinetochores. In addition, in cases of 'wait anaphase', plant SAC proteins were associated with both kinetochores and kinetochore microtubules. Unexpectedly, BUB3.1 was also found in the phragmoplast midline during the final step of cell division in plants. CONCLUSIONS/SIGNIFICANCE: We conclude that plant BUBR1, BUB3.1 and MAD2 proteins may have the SAC protein functions conserved from yeast to humans. The association of BUB3.1 with both unattached kinetochore and phragmoplast suggests that in plant, BUB3.1 may have other roles beyond the spindle assembly checkpoint itself. Finally, this study of the SAC dynamics pinpoints uncharacterised roles of this surveillance mechanism in plant cell division.

MAP65-3 microtubule-associated protein is essential for nematode-induced giant cell ontogenesis in Arabidopsis.

The infection of plants by obligate parasitic nematodes constitutes an interesting model for investigating plant cytoskeleton functions. Root knot nematodes have evolved the ability to manipulate host functions to their own advantage by redifferentiating root cells into multinucleate and hypertrophied feeding cells. These giant cells result from repeated rounds of karyokinesis without cell division. Detailed functional analyses demonstrated that Arabidopsis thaliana Microtubule-Associated Protein65-3 (MAP65-3) was essential for giant cell ontogenesis and that cytokinesis was initiated but not completed in giant cells. In developing giant cells, MAP65-3 was associated with a novel kind of cell plate-the giant cell mini cell plate-that separates daughter nuclei. In the absence of functional MAP65-3, giant cells developed but failed to fully differentiate and were eventually destroyed. These defects in giant cells impaired the maturation of nematode larvae. Thus, MAP65-3 is essential for giant cell development during root knot nematode infection. Subcellular localization of MAP65-3 and analysis of microtubule organization in the dyc283 T-DNA map65-3 mutant demonstrated that MAP65-3 played a critical role in organizing the mitotic microtubule array during both early and late mitosis in all plant organs. Here, we propose a model for the role of MAP65-3 in giant cell ontogenesis.