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Complex regulation of cell-cycle inhibitors by Fbxw7 in mouse embryonic fibroblasts

Oncogene volume 29, pages 1798–1809 (2010)Cite this article

Abstract

The F-box protein Fbxw7 (also known as Fbw7, SEL-10, hCdc4 or hAgo) mediates the ubiquitylation and thereby contributes to the degradation of proteins that positively regulate cell cycle. Conditional ablation of Fbxw7 in mouse embryonic fibroblasts (MEFs) induces cell-cycle arrest accompanied by abnormal accumulation of the intracellular domain of Notch1 (NICD1) and c-Myc. However, the molecular mechanisms by which the accumulation of NICD1 and c-Myc induces cell-cycle arrest have remained unclear. We have now examined the expression of cell-cycle inhibitors in Fbxw7-deficient MEFs and found that the abundance of p27Kip1 and p57Kip2 is paradoxically decreased. This phenomenon appears to be attributable to the accumulation of NICD1, given that it was recapitulated by overexpression of NICD1 and blocked by ablation of RBP-J. Conversely, the expression of p16Ink4a and p19ARF was increased in an NICD1-independent manner in Fbxw7-null MEFs. The increased expression of p19ARF was recapitulated by overexpression of c-Myc and abolished by ablation of c-Myc, suggesting that the accumulation of c-Myc is primarily responsible for that of p19ARF. In contrast, the upregulation of p16Ink4a appeared to be independent of c-Myc. These results indicate that cell-cycle inhibitors undergo complex regulation by the Fbxw7-mediated proteolytic system.

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Acknowledgements

We thank T Honjo for providing RbpjF/F mice; R Tsunematsu for an NICD1 plasmid; T Kitamura for pMX-puro and Plat-E cells; T Senga, Y Ono and N Kobayashi for technical assistance; and N Yanagihara, N Ishida and other laboratory members for helpful discussion and Y Akaida for help in preparation of the paper. This study was supported in part by a Grant-in-Aid and the Network Medicine Global COE Program from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Developmental Genetics, Center for Translational and Advanced Animal Research, Graduate School of Medicine, Tohoku University, Aoba-ku, Sendai, Japan

    K Masuda, Y Ishikawa & K Nakayama

  2. Department of Surgery, Graduate School of Medicine, Tohoku University, Aoba-ku, Sendai, Japan

    K Masuda & M Unno

  3. CREST, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan

    K Masuda, Y Ishikawa, I Onoyama, K I Nakayama & K Nakayama

  4. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan

    I Onoyama & K I Nakayama

  5. Department of Immunology and Oncology, National Center for Biotechnology, CNB-CSIC, Madrid, Spain

    I M de Alborán

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Correspondence to K Nakayama.

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Masuda, K., Ishikawa, Y., Onoyama, I. et al. Complex regulation of cell-cycle inhibitors by Fbxw7 in mouse embryonic fibroblasts. Oncogene 29, 1798–1809 (2010). https://doi.org/10.1038/onc.2009.469

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