Abstract
The retinoblastoma protein (pRB) negatively regulates the progression from G1 to S phase of the cell cycle, in part, by repressing E2F-dependent transcription1. pRB also possesses E2F-independent functions that contribute to cell-cycle control — for example, during pRB-mediated cell-cycle arrest pRB associates with Skp2, the F-box protein of the Skp1–Cullin–F-box protein (SCF) E3 ubiquitin ligase complex, and promotes the stability of the cyclin-dependent kinase-inhibitor p27Kip1 through an unknown mechanism2,3. Degradation of p27Kip1 is mediated by ubiquitin-dependent targeting of p27Kip1 by SCF –Skp2 (ref. 4). Here, we report a novel interaction between pRB and the anaphase-promoting complex/cyclosome (APC/C) that controls p27Kip1 stability by targeting Skp2 for ubiquitin-mediated degradation. Cdh1, an activator of APC/C, not only interacts with pRB but is also required for a pRB-induced cell-cycle arrest. The results reveal an unexpected physical convergence between the pRB tumour-suppressor protein and E3 ligase complexes, and raise the possibility that pRB may direct APC/C to additional targets during pRB-mediated cell-cycle exit.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Classon, M. & Harlow E. The retinoblastoma tumour suppressor in development and cancer. Nature Rev. Cancer 2, 910–917 (2002).
Alexander, K. & Hinds P. W. Requirement for p27(KIP1) in retinoblastoma protein-mediated senescence. Mol. Cell. Biol. 21, 3616–3631 (2001).
Ji, P. et al. An Rb-Skp2-p27 pathway mediates acute cell cycle inhibition by Rb and is retained in a partial-penetrance Rb mutant. Mol. Cell. 16, 47–58 (2004).
Amati, B. & Vlach J. Kip1 meets SKP2: new links in cell-cycle control. Nature Cell Biol. 1, E91–E93 (1999).
Weinberg, R. A. The retinoblastoma protein and cell cycle control. Cell 81, 323–330 (1995).
Thomas, D. M. et al. Terminal osteoblast differentiation, mediated by runx2 and p27KIP1, is disrupted in osteosarcoma. J. Cell. Biol. 167, 925–934 (2004).
Bloom, J. & Pagano M. Deregulated degradation of the cdk inhibitor p27 and malignant transformation. Semin. Cancer Biol. 13, 41–47 (2003).
Castro, A. et al. The anaphase-promoting complex: a key factor in the regulation of cell cycle. Oncogene 24, 314–325 (2005).
Fay, D. S., Keenan S. & Han M. fzr-1 and lin-35/Rb function redundantly to control cell proliferation in C. elegans as revealed by a nonbiased synthetic screen. Genes Dev. 16, 503–517 (2002).
Dick, F. A., Sailhamer E. & Dyson N. J. Mutagenesis of the pRB pocket reveals that cell cycle arrest functions are separable from binding to viral oncoproteins. Mol. Cell. Biol. 20, 3715–3727 (2000).
Dick, F. A. & Dyson N. J. Three regions of the pRB pocket domain affect its inactivation by human papillomavirus E7 proteins. J. Virol. 76, 6224–6234 (2002).
Isaac, C. E. et al. The retinoblastoma protein regulates pericentric heterochromatin. Mol. Cell. Biol. 26, 3659–3671 (2006).
Kraft, C. et al. The WD40 propeller domain of Cdh1 functions as a destruction box receptor for APC/C substrates. Mol. Cell. 18, 543–553 (2005).
Sorensen, C. S. et al. Nonperiodic activity of the human anaphase-promoting complex-Cdh1 ubiquitin ligase results in continuous DNA synthesis uncoupled from mitosis. Mol. Cell. Biol. 20, 7613–7623 (2000).
Wei, W. et al. Degradation of the SCF component Skp2 in cell-cycle phase G1 by the anaphase-promoting complex. Nature 428, 194–198 (2004).
Bashir, T. et al. Control of the SCF(Skp2-Cks1) ubiquitin ligase by the APC/C(Cdh1) ubiquitin ligase. Nature. 428, 190–193 (2004).
Dickins, R. A. et al. Probing tumor phenotypes using stable and regulated synthetic microRNA precursors. Nature Genet. 37, 1289–1295 (2005).
Rape, M., Reddy S. K. & Kirschner M. W. The processivity of multiubiquitination by the APC determines the order of substrate degradation. Cell 12, 89–103 (2006).
Turnell, A. S. et al. The APC/C and CBP/p300 cooperate to regulate transcription and cell-cycle progression. Nature 438, 690–695 (2005).
Stroschein, S. L. et al. Smad3 recruits the anaphase-promoting complex for ubiquitination and degradation of SnoN. Genes Dev. 15, 2822–2836 (2001).
Wan, Y., Liu X. & Kirschner M. W. The anaphase-promoting complex mediates TGF-β signaling by targeting SnoN for destruction. Mol. Cell. 8, 1027–1039 (2001).
Benevolenskaya, E. V. et al. Binding of pRB to the PHD protein RBP2 promotes cellular differentiation. Mol. Cell. 18, 623–635 (2005).
Miyake, S. et al. Cells degrade a novel inhibitor of differentiation with E1A-like properties upon exiting the cell cycle. Mol. Cell. Biol. 20, 8889–8902 (2000).
Lasorella, A. et al. Degradation of Id2 by the anaphase-promoting complex couples cell cycle exit and axonal growth. Nature 442, 471–474 (2006).
Ang, X. L. & Harper J. W. Interwoven ubiquitination oscillators and control of cell cycle transitions. Sci STKE e31 (2004).
van Roessel, P. et al. Independent regulation of synaptic size and activity by the anaphase-promoting complex. Cell 119, 707–718 (2004).
Dick, F. A. & Dyson N. J. pRB contains an E2F1-specific binding domain that allows E2F1-induced apoptosis to be regulated separately from other E2F activities. Mol. Cell. 12, 639–649 (2003).
Dignam, J. D., Lebovitz R. M. & Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 11, 1475–1489 (1983).
Imaki, H. et al. Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. Cancer Res. 63, 4607–4613 (2003).
Wirbelauer, C. et al. The F-box protein Skp2 is a ubiquitylation target of a Cul1-based core ubiquitin ligase complex: evidence for a role of Cul1 in the suppression of Skp2 expression in quiescent fibroblasts. EMBO J. 19, 5362–5375 (2000).
Zhang, H. et al. p19Skp1 and p45Skp2 are essential elements of the cyclin A-CDK2 S phase kinase. Cell 82, 915–925 (1995).
Vernell, R., Helin K. & Muller H. Identification of target genes of the p16INK4A-pRB-E2F pathway. J. Biol. Chem. 278, 46124–46137 (2003).
Acknowledgements
We thank: C. Pfleger for numerous APC/C-related plasmids; R. Watson for Myc-tagged pRB, p107, p130 plasmids; S. Lowe for U2OS shRB cell line; J. Rocco for pcDNA6/TR and pcDNA4/TO plasmids; and B. Schulman for the CMV–Myc–Skp2 plasmid. We also like to thank P. Hinds and K. Münger for helpful comments on the manuscript. This study was supported by National Institutes of Health (NIH) grant CA64402 to N.J.D and by the Massachusetts General Hospital Fund for Medical Discovery to U.K.B.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary figures S1, S2, S3, S4, S5 and S6 (PDF 531 kb)
Rights and permissions
About this article
Cite this article
Binné, U., Classon, M., Dick, F. et al. Retinoblastoma protein and anaphase-promoting complex physically interact and functionally cooperate during cell-cycle exit. Nat Cell Biol 9, 225–232 (2007). https://doi.org/10.1038/ncb1532
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb1532
This article is cited by
-
Cell cycle regulation: p53-p21-RB signaling
Cell Death & Differentiation (2022)
-
CDK4/6 inhibitors downregulate the ubiquitin-conjugating enzymes UBE2C/S/T involved in the ubiquitin–proteasome pathway in ER + breast cancer
Clinical and Translational Oncology (2022)
-
Theileria parasites subvert E2F signaling to stimulate leukocyte proliferation
Scientific Reports (2020)
-
Time varying causal network reconstruction of a mouse cell cycle
BMC Bioinformatics (2019)
-
Mechanisms for the temporal regulation of substrate ubiquitination by the anaphase-promoting complex/cyclosome
Cell Division (2019)