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Degradation of the SCF component Skp2 in cell-cycle phase G1 by the anaphase-promoting complex

Nature volume 428, pages 194–198 (2004)Cite this article

Abstract

Cell-cycle transitions are driven by waves of ubiquitin-dependent degradation of key cell-cycle regulators. SCF (Skp1/Cullin/F-box protein) complexes and anaphase-promoting complexes (APC) represent two major classes of ubiquitin ligases whose activities are thought to regulate primarily the G1/S and metaphase/anaphase cell-cycle transitions, respectively1,2. The major target of the Skp1/Cul1/Skp2 (SCFSKP2) complex is thought to be the Cdk inhibitor p27 during S phase, whereas the principal targets for the APC are thought to be involved in chromatid separation (securin) and exit from mitosis (cyclin B). Although the role of the APC in mitosis is relatively clear, there is mounting evidence that APCs containing Cdh1 (APCCDH1) also have a function in the G1 phase of the cell cycle2,3. Here, we show that the F-box protein Skp2 is polyubiquitinated, and hence earmarked for destruction, by APCCDH1. As a result, accumulation of SCFSKP2 requires prior inactivation of APCCDH1. These findings provide an insight into the orchestration of SCF and APC activities during cell-cycle progression, and into the involvement of the APC in G1.

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Figure 1: D-box-dependent degradation of Skp2 in G1 extracts.
Figure 2: Polyubiquitination and destruction of Skp2 by APC in vitro.
Figure 3: Cdh1 is necessary and sufficient for downregulation of Skp2 in G1.
Figure 4: D-box-dependent turnover of Skp2 in G1.

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Acknowledgements

We thank W. Harper, J. Lukas and M. Meyerson for critical reading of the manuscript; J. Lukas for reagents; M. Pagano for sharing unpublished data; and members of the Kirschner and Kaelin Laboratories for useful discussions. W.G.K. is a Howard Hughes Medical Institute Investigator. This work is supported in part by NIH grants to M.W.K. and W.G.K.

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Author notes

  1. Wenyi Wei and Nagi G. Ayad: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Medical Oncology, Dana–Farber Cancer Institute and Brigham and Women's Hospital,

    Wenyi Wei, Guo-Jun Zhang & William G. Kaelin Jr

  2. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, 02115, USA

    Nagi G. Ayad, Yong Wan & Marc W. Kirschner

  3. Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, 02115, USA

    William G. Kaelin Jr

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  1. Wenyi Wei
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Correspondence to Marc W. Kirschner or William G. Kaelin Jr.

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Supplementary information

Supplementary figure 1

Binding of Cdh1 mutants to Skp2. (PDF 1105 kb)

Supplementary figure 2

Binding of Skp2 mutants to Cdh1. (PDF 787 kb)

Supplementary figure 3

Binding of Skp2 and Skp2Δdb to Cdh1 in vivo. (PDF 610 kb)

Supplementary figure 4

Immunoblot analysis of HeLa cells transfected with the indicated siRNA, synchronized by growth in nocodazole, and then released for the indicated periods of time. (PDF 473 kb)

Supplementary figure legends (DOC 21 kb)

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Wei, W., Ayad, N., Wan, Y. et al. Degradation of the SCF component Skp2 in cell-cycle phase G1 by the anaphase-promoting complex. Nature 428, 194–198 (2004). https://doi.org/10.1038/nature02381

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