Targeting of Fzr/Cdh1 for timely activation of the APC/C at the centrosome during mitotic exit
Abstract
A multi-subunit ubiquitin ligase, the anaphase-promoting complex/cyclosome (APC/C), regulates critical cellular processes including the cell cycle. To accomplish its diverse functions, APC/C activity must be precisely regulated in time and space. The interphase APC/C activator Fizzy-related (Fzr or Cdh1) is localized at centrosomes in animal cells. However, neither the mechanism of its localization nor its importance is clear. Here we identify the centrosome component Spd2 as a major partner of Fzr in Drosophila. The localization of Fzr to the centriole during interphase depends on direct interaction with Spd2. By generating Spd2 mutants unable to bind Fzr, we show that centrosomal localization of Fzr is essential for optimal APC/C activation towards its centrosomal substrate Aurora A. Finally, we show that Spd2 is also a novel APC/C^(Fzr) substrate. Our study is the first to demonstrate the critical importance of distinct subcellular pools of APC/C activators in the spatiotemporal control of APC/C activity.
Additional Information
© The Author(s) 2016. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 21 December 2015. Accepted 16 July 2016. Published 25 August 2016. This work is supported by Cancer Research UK Career Development Fellowship to Y.K., a Project Grant to Y.K. and D.M.G. from BBSRC, and a Medical Research Council (MRC) Project Grant to H.Y. Y.K. thanks the Japan Society for the Promotion of Science for the Postdoctoral Fellowship for Research Abroad and T.M. thanks the European Commission for Marie Skłodowska-Curie actions individual fellowship. We thank Nicola Lawrence and Alex Sossick at the Imaging facility and David Simpson at the Frog facility at Gurdon Institute for assistance with 3D-SIM and egg extract preparation, respectively, the Fly facility and the media service at Department of Genetics for their service, and Drosophila research community for sharing the reagents. We thank all the Kimata and Glover lab members for discussion and cooperation. Y.K. thanks Iain Hagan and Alexis Braun for feedback on the manuscript and Jens Januschke, Renata Basto and Paul Conduit for discussion and comments on the manuscript. Francesco Meghini, Torcato Martins and Xavier Tait: These authors contributed equally to this work. Author Contributions: Y.K. supervised the whole project, designed and performed experiments for Figs 1a–c, 2b,c, 4d,i, 5c,d, and wrote this manuscript with a help of F.M., T.M. and D.M.G. F.M. designed and conducted the experiments for Figs 3a–d,g, 5g,h, 8c–e and in vitro destruction assays and most of the quantitation and the statistical analyses of the data. T.M. designed and performed the experiments in all the live imaging experiments and the larval brain analysis. X.T. designed and performed the experiments in Figs 1d–f, 2a, 3b,g and 5a,b. K.F. designed and performed the in vitro ubiquitination assay with a help of H.Y. D.M.G. helped Y.K. in project supervision. Data availability: The data that support the findings of this study are available from the corresponding author upon request. The authors declare no competing financial interests.Attached Files
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Additional details
- PMCID
- PMC5007356
- Eprint ID
- 104875
- Resolver ID
- CaltechAUTHORS:20200807-170354727
- Cancer Research UK
- Biotechnology and Biological Sciences Research Council (BBSRC)
- Medical Research Council (UK)
- Japan Society for the Promotion of Science (JSPS)
- Marie Curie Fellowship
- Created
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2020-08-10Created from EPrint's datestamp field
- Updated
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2021-11-16Created from EPrint's last_modified field