Abstract
HIPK2 is a eukaryotic Serine–Threonine kinase that controls cellular proliferation and survival in response to exogenous signals. Here, we show that the human transcription factor ZBTB4 is a new target of HIPK2. The two proteins interact in vitro, colocalize and associate in vivo, and HIPK2 phosphorylates several conserved residues of ZBTB4. Overexpressing HIPK2 causes the degradation of ZBTB4, whereas overexpressing a kinase-deficient mutant of HIPK2 has no effect. The chemical activation of HIPK2 also decreases the amount of ZBTB4 in cells. Conversely, the inhibition of HIPK2 by drugs or by RNA interference causes a large increase in ZBTB4 levels. This negative regulation of ZBTB4 by HIPK2 occurs under normal conditions of cell growth. In addition, the degradation is increased by DNA damage. These findings have two consequences. First, we have recently shown that ZBTB4 inhibits the transcription of p21. Therefore, the activation of p21 by HIPK2 is two-pronged: stimulation of the activator p53, and simultaneous repression of the inhibitor ZBTB4. Second, ZBTB4 is also known to bind methylated DNA and repress methylated sequences. Consequently, our findings raise the possibility that HIPK2 might influence the epigenetic regulation of gene expression at loci that remain to be identified.
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Acknowledgements
We thank the following colleagues for reagents: Nelson Dusetti, Roger Tsien, Phillip James, Jacques Camonis, Silvia Soddu. We are especially grateful to Gabriella d′Orazi for the HIPK2 knockdown cells, and to Patricia Le Baccon for her help. DY was supported by postdoctoral fellowships from the Curie Institute, and from Association pour la Recherche contre le Cancer. RPT was supported by a fellowship from Centre National de la Recherche Scientifique (CNRS). The Defossez lab is supported by CNRS (programme ATIP and programme PICS France-Russie), by Institut National du Cancer (ATIP Plus), by Association pour la Recherche contre le Cancer (grants no 3727 and no 4859), and by Ligue contre le Cancer. The work of MLS is supported by the Deutsche Forschungsgemeinschaft (DFG).
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)
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Yamada, D., Pérez-Torrado, R., Filion, G. et al. The human protein kinase HIPK2 phosphorylates and downregulates the methyl-binding transcription factor ZBTB4. Oncogene 28, 2535–2544 (2009). https://doi.org/10.1038/onc.2009.109
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DOI: https://doi.org/10.1038/onc.2009.109
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