Abstract
Protein phosphotase Cdc14 (Cell division cycle gene 14) is a key regulator of late mitotic events in Saccharomyces cerevisiae. However the function of human Cdc14 (HsCdc14A & B) and its regulatory network are still elusive. In this study, we identified a new partner of HsCdc14A named Brap2 (BRCA1 associated protein 2) using yeast two-hybrid screening assay. The interaction between these two proteins is confirmed by co-immunoprecipitation in human HEK 293T cells. Brap2 co-localizes with HsCdc14A on mitotic spindle poles and over-expression of Brap2 causes multiple spindle poles. Furthermore, we found that Brap2, which has intrinsic RING domain dependent E3 ligase activity, facilitates HsCdc14A Lys-63 linked ubiquitin modification, indicating that Brap2 may be the ubiquitin E3 Ligase of HsCdc14A. Our findings imply that Brap2 plays a significant role in cell cycle regulation besides its facilitation of HsCdc14A ubiquitination.
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Acknowledgments
We thank J. Lukas (Institute of Cancer Biology, Copenhagen, Denmark) for the human HsCdc14A cDNA (GenBankTM Accession NO. Q5VUH9), M.A. White (Department of Cell Biology, Texas, USA) for Brap2 cDNA (GenBankTM Accession NO. Q7Z569), and Yixian Zheng (Department of Embryology, MD, USA) for HA-Ub (Wt, K48R and K63R) plasmids. This work was supported by Chinese 973 Project Grant (2002CB713703), the Key Project of National Natural Science Foundation of China (30230380).
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Chen, JS., Hu, HY., Zhang, S. et al. Brap2 facilitates HsCdc14A Lys-63 linked ubiquitin modification. Biotechnol Lett 31, 615–621 (2009). https://doi.org/10.1007/s10529-009-9914-7
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DOI: https://doi.org/10.1007/s10529-009-9914-7