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Ubiquitin hydrolase Dub3 promotes oncogenic transformation by stabilizing Cdc25A

Nature Cell Biology volume 12, pages 400–406 (2010)Cite this article

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Abstract

The dual specificity (Tyr/Thr) phosphatase Cdc25A activates cyclin-dependent kinases (Cdks) to promote cell-cycle progression and has significant oncogenic potential1. Cdc25A protein levels are regulated tightly in normal tissues, but many human cancers overexpress Cdc25A. The underlying mechanism for overexpression has been enigmatic2. Here we show that Cdc25A is stabilized by the ubiquitin hydrolase Dub3. Upon binding Cdc25A, Dub3 removes the polyubiquitin modifications that mark Cdc25A for proteasomal degradation. Dub3 knockdown in cells increased Cdc25A ubiquitylation and degradation, resulting in reduced Cdk/Cyclin activity and arrest at G1/S and G2/M phases of the cell cycle. In contrast, acute Dub3 overexpression produced a signature response to oncogene induction: cells accumulated in S and G2 because of replication stress, and activated a DNA damage response. Dub3 also transformed NIH-3T3 cells and cooperated with activated H-Ras to promote growth in soft agar. Importantly, we show that Dub3 overexpression is responsible for an abnormally high level of Cdc25A in a subset of human breast cancers. Moreover, Dub3 knockdown significantly retarded the growth of breast tumour xenografts in nude mice. As a major regulator of Cdc25A, Dub3 is an example of a transforming ubiquitin hydrolase that subverts a key component of the cell cycle machinery.

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Figure 1: Dub3 regulates Cdc25A stability.
Figure 2: Dub3 knockdown decreases Cdc25A and causes cell-cycle arrest.
Figure 3: Constitutive Dub3 expression activates a DNA damage response and apoptosis.
Figure 4: Effect of Cdc25A, APC/Cdh1 or SCFβTrCP knockdown, or Myc–Cdc25A overexpression on cells overexpressing Dub3.
Figure 5: Dub3 has oncogenic potential.

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Acknowledgements

We thank K. Newton for editorial assistance, J.Z. Torres for the H2B/Tub-U2OS cells; J. Cupp and L. Gilmour for technical assistance; I.E.Wertz, H. Maecker, E.W. Verschuren and members of the Dixit Lab for helpful discussions.

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Authors and Affiliations

  1. Physiological Chemistry, 1 DNA Way, South San Francisco, 94080, California, USA

    Yaron Pereg, Karen M. O'Rourke, Anwesha Dey & Vishva M. Dixit

  2. Cancer Pathways and Targets, 1 DNA Way, South San Francisco, 94080, California, USA

    Bob Y. Liu

  3. Pathology, Genentech, 1 DNA Way, South San Francisco, 94080, California, USA

    Meredith Sagolla, Laszlo Komuves & Dorothy M. French

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  1. Yaron Pereg
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Contributions

V.M.D directed the study; Y.P., with assistance from K.M.O. and A.D., conducted all biochemical experiments; B.Y.L., with assistance from Y.P., ran the xenograft studies; M.S. and L.K. performed the microscopy analyses; D.M.F generated the immunohistochemical data.

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Correspondence to Vishva M. Dixit.

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All authors are employees of Genentech.

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Pereg, Y., Liu, B., O'Rourke, K. et al. Ubiquitin hydrolase Dub3 promotes oncogenic transformation by stabilizing Cdc25A. Nat Cell Biol 12, 400–406 (2010). https://doi.org/10.1038/ncb2041

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