Abstract
The evolutionarily conserved WD40 protein budding uninhibited by benzimidazole 3 (BUB3) is known for its function in spindle assembly checkpoint control. In the model plant Arabidopsis thaliana, nearly identical BUB3;1 and BUB3;2 proteins decorated the phragmoplast midline through interaction with the microtubule-associated protein MAP65-3 during cytokinesis. BUB3;1 and BUB3;2 interacted with the carboxy-terminal segment of MAP65-3 (but not MAP65-1), which harbours its microtubule-binding domain for its post-mitotic localization. Reciprocally, BUB3;1 and BUB3;2 also regulated MAP65-3 localization in the phragmoplast by enhancing its microtubule association. In the bub3;1 bub3;2 double mutant, MAP65-3 localization was often dissipated away from the phragmoplast midline and abolished upon treatment of low doses of the cytokinesis inhibitory drug caffeine that were tolerated by the control plant. The phragmoplast microtubule array exhibited uncoordinated expansion pattern in the double mutant cells as the phragmoplast edge reached the parental plasma membrane at different times in different areas. Upon caffeine treatment, phragmoplast expansion was halted as if the microtubule array was frozen. As a result, cytokinesis was abolished due to failed cell plate assembly. Our findings have uncovered a novel function of the plant BUB3 in MAP65-3-dependent microtubule reorganization during cytokinesis.
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07 August 2018
In the version of this Article originally published, the affiliation for author Yuh-Ru Julie Lee was incorrect; the correct affiliation is ‘2Department of Plant Biology, College of Biological Sciences, University of California, Davis, CA, USA’. This has now been amended in all versions of the Article.
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Acknowledgements
We thank members of the Liu laboratory for critical comments on the work. Special thanks go to S. P. Dinesh-Kumar for the BiFC vectors and T. Nakagawa at Shimane University in Japan for the pGWB plasmids, and S. Bednarek for the DRP1A antibody. This work was supported by the US National Science Foundation under the grant MCB-1412509 to B.L. and Y.-R.J.L. H.Z. was supported by a fellowship from the China Scholarship Council (no. 201406305055), the Fundamental Research Funds for the Central Universities (no. 2452018155) and the 111 Project from the Ministry of Education of China (no. B07049). Any opinions, findings and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the funding agency.
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Y.-R.J.L. and B.L. conceived the project and designed the experiments. H.Z., X.D., B.S., S.L.V. and Y.-R.J.L. performed the experiments. H.Z., X.D., Z.K., H.L., Y.-R.J.L. and B.L. analysed the data and interpreted the results. Y.-R.J.L. and B.L. wrote the manuscript with inputs from other authors.
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Supplementary Tables 1 and 2, and Supplementary Figures 1 and 2
Supplementary Video 1
BUB3;1-GFP localization during mitotic cell division in A. thaliana. This experiment was repeated independently five times with similar results.
Supplementary Video 2
MT reorganization during mitotic cell division in a control cell (left) and a bub3;1 bub3;2 double mutant cell (right) in A. thaliana under undisturbed conditions. This experiment was repeated independently three times with similar results.
Supplementary Video 3
MT reorganization during mitotic cell division in a control cell in A. thaliana after being exposed to 1 mM caffeine this experiment was repeated independently three times with similar results.
Supplementary Video 4
MT reorganization during mitotic cell division in a bub3;1 bub3;2 double mutant cell in A. thaliana after being exposed to 1 mM caffeine. This experiment was repeated independently three times with similar results.
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Zhang, H., Deng, X., Sun, B. et al. Role of the BUB3 protein in phragmoplast microtubule reorganization during cytokinesis. Nature Plants 4, 485–494 (2018). https://doi.org/10.1038/s41477-018-0192-z
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DOI: https://doi.org/10.1038/s41477-018-0192-z
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