Abstract
During cell division, chromosome segregation is orchestrated by the interaction of spindle microtubules with the centromere. A dramatic remodeling of interpolar microtubules into an organized central spindle between the separating chromatids is required for the initiation and execution of cytokinesis. Central spindle organization requires mitotic kinesins, the chromosomal passenger protein complex, and microtubule bundling protein PRC1. PRC1 is phosphorylated by Cdc2 at Thr470 and Thr481 during mitosis. However, the functional relevance of PRC1 phosphorylation at Thr470 has remained elusive. Here we show that expression of the non-phosphorylatable mutant PRC1T470A but not the phospho-mimicking mutant PRC1T470E causes aberrant organization of the central spindle. Immunoprecipitation experiment indicates that both PRC1T470A and PRC1T470E mutant proteins associate with wild-type PRC1, suggesting that phosphorylation of Thr470 does not alter PRC1 self-association. In addition, in vitro co-sedimentation experiment showed that PRC1 binds to microtubule independent of the phosphorylation state of Thr470. Gel-filtration experiment suggested that phosphorylation of Thr470 promotes oligomerization of PRC1. Given the fact that prevention of the Thr470 phosphorylation inhibits PRC1 oligomerization in vitro and causes an aberrant organization of central spindle in vivo, we propose that this phosphorylation-dependent PRC1 oligomerization ensures that central spindle assembly occurs at the appropriate time in the cell cycle.
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Acknowledgements
We are grateful to Dr Wei Jiang for gift of reagents. We also thank the members of our group for helpful discussion during the course of this study. This work was supported by National Natural Science Foundation of China (39925018, 90508002 and 30121001), Chinese Academy of Science (KSCX1-R65 and RSCX2-H10), National Basic Research Program of China (973 project, 2002CB713700), American Cancer Society (RPG-99-173-01), a Gcc Breast Cancer Research award and National Institutes of Health grants DK56292 and CA89019 to XY (a GCC Eminent Scholar) and NS36194 (JW).
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Fu, C., Yan, F., Wu, F. et al. Mitotic phosphorylation of PRC1 at Thr470 is required for PRC1 oligomerization and proper central spindle organization. Cell Res 17, 449–457 (2007). https://doi.org/10.1038/cr.2007.32
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DOI: https://doi.org/10.1038/cr.2007.32
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