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p55CDC/hCDC20 is associated with BUBR1 and may be a downstream target of the spindle checkpoint kinase

Oncogene volume 19, pages 4557–4562 (2000)Cite this article

Abstract

Eukaryotic cells have evolved a mechanism that delays the progression of mitosis until condensed chromosomes are properly positioned on the mitotic spindle. We have been studying genes that regulated the spindle checkpoint in human cells. Enforced expression of human BUBR1, but not a BUBR1 mutant allele, enhances accumulation of mitotic cells. Yeast two-hybrid system and GST-pull-down analyses show that p55CDC/hCdc20, a protein known to link spindle checkpoint components such as MAD2 to anaphase promoting complex (APC), interacts with BUBR1. In addition, p55CDC is capable of pulling down BUBR1 in sf-9 cells infected with both p55CDC and His6-BUBR1 recombinant baculoviruses but not in the cells infected with p55CDC baculoviruses or with the baculoviral vector alone. Moreover, immunoprecipitation followed by Western blot analyses confirmed that native p55CDC is associated with BUBR1 in HeLa cells. Spindle checkpoint activation by nocodazole treatment enhances the association between p55CDC and His6-BUBR1. In nocodazole-arrested mitotic cells, both CDC16 and hyperphosphorylated CDC27, two APC components, preferentially associate with His6-BUBR1 resins, but not the control resins. Furthermore, BUBR1 phosphorylates p55CDC in vitro, and the phosphorylation of p55CDC by BUBR1 appears to be correlated with spindle checkpoint activation. Together, our studies strongly suggest that BUBR1 may target APC via p55CDC.

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Acknowledgements

We are grateful to Philip Hieter and Robert Benezra for providing us with anti-CDC16 and anti-CDC27 antibodies, respectively. We thank Andrew Hoyt for valuable suggestions and helpful discussions. This work was supported by the Public Service Award CA74299.

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Author notes

  1. Huiyun Wu, Zhengdao Lan and Wenqing Li: H Wu, Z Lan and W Li contributed equally to the work reported in the manuscript

Authors and Affiliations

  1. American Health Foundation, Valhalla, New York, 10595, NY, USA

    Huiyun Wu & Wei Dai

  2. Amgen, Inc. Thousand Oaks, California, USA

    Jasminder Weinstein

  3. Hematology-Oncology Division, Department of Pediatrics and Pathology and Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angeles, California, USA

    Kathleen M Sakamoto

  4. Division of Hematology-Oncology, University of Cincinnati, Cincinnati, Ohio, USA

    Zhengdao Lan, Wenqing Li & Shechao Wu

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  1. Huiyun Wu
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Wu, H., Lan, Z., Li, W. et al. p55CDC/hCDC20 is associated with BUBR1 and may be a downstream target of the spindle checkpoint kinase. Oncogene 19, 4557–4562 (2000). https://doi.org/10.1038/sj.onc.1203803

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