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DNA-PK phosphorylation sites on Oct-1 promote cell survival following DNA damage

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Abstract

Octamer transcription factor-1 (Oct-1) has recently been shown to function as a stress sensor that promotes cell survival subsequent to DNA damage. Here, we show that the survival signal imparted by Oct-1 following exposure to ionizing radiation (IR) is dependent upon DNA-dependent protein kinase (DNA-PK)-dependent phosphorylation of a cluster of 13 specific ser/thr residues within the N-terminal transcriptional regulatory domain of Oct-1. Although IR treatment did not affect the recruitment of Oct-1 to the histone H2B promoter, the recruitment of RNA polymerase II, TATA-binding protein and histone H4 acetylation were strongly reduced, consistent with a decrease in Oct-1 transcriptional regulatory potential following IR exposure. Ser/Thr-Ala substitution of 13 sites present in Oct-1 transcriptional regulatory domain eliminated Oct-1 phosphorylation subsequent to IR exposure. Further, these substitutions prevented Oct-1 from rescuing the survival of IR-treated Oct-1/ murine embryonic fibroblasts, providing a direct link between DNA-PK-dependent phosphorylation and the contribution of Oct-1 to cell survival. These results implicate Oct-1 as a primary effector in a DNA-PK-dependent cell survival pathway that is activated by double-stranded DNA breaks.

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Acknowledgements

We are grateful to Winship Herr for providing the pCGNOct-1 plasmid and Shigemi Matsuyama for the Ku−/− and +/+ MEFs. This work was supported by an operating grant from the Cancer Research Society Inc. (CRS) to RJGH and CS-P. RJGH is an investigator of the Canadian Institutes for Health Research.

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Author notes

  1. C Schild-Poulter

    Present address: Cell Biology Research Group, Robarts Research Institute, PO Box 5015, 100, Perth Drive, London, Ontario, Canada, N6A 5K8

  2. D Tantin

    Present address: Department of Pathology, University of Utah School of Medicine, EEJB-565, Room 5700B, 15 No. Medical Drive East, Salt Lake City, UT, 84112-5650, USA

Authors and Affiliations

  1. Department of Medicine, The Ottawa Health Research Institute, University of Ottawa, Ottawa, Ontario, Canada

    C Schild-Poulter, A Shih, N C Yarymowich, S Soubeyrand & R J G Haché

  2. Department of Biochemistry, Microbiology and Immunology, The Ottawa Health Research Institute, University of Ottawa, Ottawa, Ontario, Canada

    R J G Haché

  3. Center for Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA

    D Tantin & P A Sharp

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  1. C Schild-Poulter
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Correspondence to C Schild-Poulter or R J G Haché.

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

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Schild-Poulter, C., Shih, A., Tantin, D. et al. DNA-PK phosphorylation sites on Oct-1 promote cell survival following DNA damage. Oncogene 26, 3980–3988 (2007). https://doi.org/10.1038/sj.onc.1210165

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