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Fbxw7/Cdc4 is a p53-dependent, haploinsufficient tumour suppressor gene

Abstract

The FBXW7/hCDC4 gene encodes a ubiquitin ligase implicated in the control of chromosome stability1. Here we identify the mouse Fbxw7 gene as a p53-dependent tumour suppressor gene by using a mammalian genetic screen for p53-dependent genes involved in tumorigenesis. Radiation-induced lymphomas from p53+/- mice, but not those from p53-/- mice, show frequent loss of heterozygosity and a 10% mutation rate of the Fbxw7 gene. Fbxw7+/- mice have greater susceptibility to radiation-induced tumorigenesis, but most tumours retain and express the wild-type allele, indicating that Fbxw7 is a haploinsufficient tumour suppressor gene. Loss of Fbxw7 alters the spectrum of tumours that develop in p53 deficient mice to include a range of tumours in epithelial tissues such as the lung, liver and ovary. Mouse embryo fibroblasts from Fbxw7-deficient mice, or wild-type mouse cells expressing Fbxw7 small interfering RNA, have higher levels of Aurora-A kinase, c-Jun and Notch4, but not of cyclin E. We propose that p53-dependent loss of Fbxw7 leads to genetic instability by mechanisms that might involve the activation of Aurora-A, providing a rationale for the early occurrence of these mutations in human cancers.

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Figure 1: LOH and mutation analysis of radiation-induced lymphoma.
Figure 2: Relationship between Fbxw7 and p53.
Figure 3: Radiation-induced tumorigenesis in p53-deficient and Fbxw7-deficient mice.
Figure 4: Development of multiple tumour types in p53+/-Fbxw7+/- mice.

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Acknowledgements

We thank the staff of the CRUK Beatson Institute and UCSF Comprehensive Cancer Center animal house for animal husbandry; L. Heath (Laboratory Animal Resource Center, UCSF) for analysing the histological slides from tumors; and J. Hoh (Laboratories of Statistical Genetics and Cancer Biology of The Rockefeller University, New York, USA) for providing the p53MH algorithm program. The UCSF Cancer Center Genome Core was essential for the sequencing and study design of the Taqman assay. These studies were initially supported by the Commission of the European Communities and the Cancer Research Campaign (UK), and subsequently by an NCI grant and a DOE grant to A.B. J.H.M. is the recipient of a Leukemia & Lymphoma Society Fellowship. J.P.L. has a Fellowship from the ‘Ministerio de Educacion y Ciencia’ of Spain. A.B. is the recipient of the Barbara Bass Bakar Chair in Cancer Genetics.

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Correspondence to Allan Balmain.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Notes

Contains Supplementary Methods and legends for Supplementary Figures S1–S5. (DOC 32 kb)

Supplementary Figure S1a

LOH detected by microsatellite analysis, distribution of microsatellite markers used to detect LOH by PCR. (PDF 6 kb)

Supplementary Figure S1b

LOH detected by microsatellite analysis, representative LOH pattern (PDF 77 kb)

Supplementary Figure S2

Identification of mutations in Fbxw7 in lymphomas from p53+/- mice. (PDF 8 kb)

Supplementary Figure S3

FACS analysis of aneuploidy in MEFs expressing Fbxw7 RNAi (PDF 82 kb)

Supplementary Figure S4

Expression of p53 detected by Western blotting in MEFs with or without Fbxw7 RNAi. (PDF 23 kb)

Supplementary Figure S5

Proposed scheme for the sequence of events in radiation induced lymphoma in p53 deficient mice (PDF 6 kb)

Supplementary Table 1

Putative p53 DNA-responsive elements within 10 Kb promoter region of mouse Fbxwα, β and γ. (PDF 7 kb)

Supplementary Tables 2–5

Supplementary Table 2: list of D3MA markers. Supplementary Table 3: primer for DNA sequencing Fbxw7 . Supplementary Table 4: primer for RT-PCR and sequencing Fbxw7. (PDF 86 kb)

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Mao, JH., Perez-losada, J., Wu, D. et al. Fbxw7/Cdc4 is a p53-dependent, haploinsufficient tumour suppressor gene. Nature 432, 775–779 (2004). https://doi.org/10.1038/nature03155

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