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Notch-dependent cell cycle arrest and apoptosis in mouse embryonic fibroblasts lacking Fbxw7

Oncogene volume 27, pages 6164–6174 (2008)Cite this article

Abstract

The F-box protein Fbxw7 mediates the ubiquitylation and consequent degradation of proteins that regulate cell cycle progression, including cyclin E, c-Myc, c-Jun and Notch. Moreover, certain human cancer cell lines harbor loss-of-function mutations in FBXW7 that result in excessive accumulation of Fbxw7 substrates, implicating Fbxw7 in tumor suppression. To elucidate the physiological function of Fbxw7, we conditionally ablated Fbxw7 in mouse embryonic fibroblasts (MEFs). Unexpectedly, loss of Fbxw7 induced cell cycle arrest and apoptosis that were accompanied by abnormal accumulation of the intracellular domain of Notch1 (NICD1). Forced expression of NICD1 in wild-type MEFs recapitulated the phenotype of the Fbxw7-deficient (Fbxw7Δ/Δ) MEFs. Conversely, deletion of Rbpj normalized the phenotype of Fbxw7Δ/Δ MEFs, indicating that this phenotype is dependent on the Notch1–RBP-J signaling pathway. Deletion of the p53 gene prevented cell cycle arrest but not the induction of apoptosis in Fbxw7Δ/Δ cells. These observations suggest that Fbxw7 does not function as an oncosuppressor in MEFs. Instead, it promotes cell cycle progression and cell survival through degradation of Notch1, with loss of Fbxw7 resulting in NICD1 accumulation, cell cycle arrest and apoptosis.

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Acknowledgements

We thank T Honjo for RbpjF/F mice; R Tsunematsu for an NICD1 plasmid; T Kitamura for pMX-puro and Plat-E cells; Y Ono and N Kobayashi for technical assistance; N Ishida and other laboratory members for helpful discussion. This study was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and by the 21st Century Center of Excellence Program.

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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

    Y Ishikawa & K Nakayama

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

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

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

    I Onoyama & K I Nakayama

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  1. Y Ishikawa
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Correspondence to K Nakayama.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Ishikawa, Y., Onoyama, I., Nakayama, K. et al. Notch-dependent cell cycle arrest and apoptosis in mouse embryonic fibroblasts lacking Fbxw7. Oncogene 27, 6164–6174 (2008). https://doi.org/10.1038/onc.2008.216

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