Abstract
Fbw7 and Cdh1 are substrate-recognition subunits of the SCF- and APC-type E3 ubiquitin ligases, respectively. There is emerging evidence suggesting that both Fbw7 and Cdh1 function as tumor suppressors by targeting oncoproteins for destruction. Loss of Fbw7, but not Cdh1, is frequently observed in various human tumors. However, it remains largely unknown how Fbw7 mechanistically functions as a tumor suppressor and whether there is a signaling crosstalk between Fbw7 and Cdh1. Here, we report that Fbw7-deficient cells not only display elevated expression levels of SCFFbw7 substrates, including cyclin E, but also have increased expression of various APCCdh1 substrates. We further defined cyclin E as the critical signaling link by which Fbw7 governs APCCdh1 activity, as depletion of cyclin E in Fbw7-deficient cells results in decreased expression of APCCdh1 substrates to levels comparable to those in wild-type (WT) cells. Conversely, ectopic expression of cyclin E recapitulates the aberrant APCCdh1 substrate expression observed in Fbw7-deficient cells. More importantly, 4A-Cdh1 that is resistant to Cdk2/cyclin E-mediated phosphorylation, but not WT-Cdh1, reversed the elevated expression of various APCCdh1 substrates in Fbw7-deficient cells. Overexpression of 4A-Cdh1 also resulted in retarded cell growth and decreased anchorage-independent colony formation. Altogether, we have identified a novel regulatory mechanism by which Fbw7 governs Cdh1 activity in a cyclin E-dependent manner. As a result, loss of Fbw7 can lead to aberrant increase in the expression of both SCFFbw7 and APCCdh1 substrates. Our study provides a better understanding of the tumor suppressor function of Fbw7, and suggests that Cdk2/cyclin E inhibitors could serve as effective therapeutic agents for treating Fbw7-deficient tumors.
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Acknowledgements
We would like to thank Shavali Shaik and Zhiwei Wang for critical reading of the manuscript, and William Hahn for providing reagents. WW is an MLSC New Investigator, ACS Research Scholar, LLS Research Scholar and DOD Prostate Cancer Program New Investigator, and AWL is an NRSA T32 postdoctoral fellow. This work was supported in part by the NIH grants (WW, GM089763 and GM094777; YS, CA118762 and CA156744).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
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Supplementary information, Figure S1
Depletion of Fbw7 leads to aberrant elevation of various Cdh1 downstream substrates. (PDF 180 kb)
Supplementary information, Figure S2
Cyclin E plays an important role in mediating the ability of Fbw7 to regulate Cdh1 substrate abundance. (PDF 111 kb)
Supplementary information, Figure S3
The Fbw7/Cyclin E signaling axis negatively regulates the stability of various Cdh1 substrates. (PDF 215 kb)
Supplementary information, Figure S4
Phosphorylation of Cdh1 by Cdk2/Cyclin E at multiple sites results in the inactivation of the APC/Cdh1 E3 ubiquitin ligase. (PDF 128 kb)
Supplementary information, Figure S5
Cyclin E-dependent suppression of Cdh1 E3 ligase activity contributes to the tumor suppressor function of Fbw7. (PDF 312 kb)
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Lau, A., Inuzuka, H., Fukushima, H. et al. Regulation of APCCdh1 E3 ligase activity by the Fbw7/cyclin E signaling axis contributes to the tumor suppressor function of Fbw7. Cell Res 23, 947–961 (2013). https://doi.org/10.1038/cr.2013.67
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DOI: https://doi.org/10.1038/cr.2013.67
