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Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression

Oncogene volume 26, pages 3541–3550 (2007)Cite this article

Abstract

The proto-oncoproteins ETS1 and growth factor independent-1 (GFI1) are implicated in cell growth and differentiation in various types of cells, and their deregulated expression is involved in malignant transformation. Here, we report that ETS1 and GFI1 interact and affect gene expression through their cross-talk. Co-immunoprecipitation analyses and glutathione-S-transferase pull-down assays revealed that ETS1 bound directly to GFI1 via its Ets domain, and GFI1 bound to ETS1 via its zinc-finger domain. Luciferase (Luc) assays using artificial reporters showed that GFI1 repressed ETS1-mediated transcriptional activation and ETS1 repressed GFI1-mediated transcriptional activation, in a dose-dependent manner. However, in the Bax promoter where the Ets- and Gfi-binding sites (EBS and GBS) are adjacent, ETS1 and GFI1 cooperatively reduced activation. Site-directed mutagenesis on the EBS and GBS of the Bax promoter showed that both binding sites were necessary for full repression. Chromatin immunoprecipitation analyses confirmed that an ETS1–GFI1 complex formed on the Bax promoter even when either EBS or GBS was mutated. Introduction of small interfering RNA against ETS1 and/or GFI1 enhanced endogenous Bax gene expression. Our results suggest that the interaction between ETS1 and GFI1 facilitates their binding to specific sites on the Bax promoter and represses Bax expression in vivo.

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Acknowledgements

This work was supported by a Grant-in-Aid to TO from the Ministry of Education, Science and Culture of Japan (no. 17591019). Financial supports to TO from Dr J Akiyama, OBGYN Akiyama Memorial Hospital, Hakodate and Dr K Watanabe, Watanabe Clinic, Shizuoka Japan are also acknowledged.

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Authors and Affiliations

  1. Department of Cell Genetics, Sasaki Institute, Chiyoda-ku, Tokyo, Japan

    Y Nakazawa, M Suzuki, N Manabe, T Yamada, F Kihara-Negishi, T Sakurai & T Oikawa

  2. Division of Organic and Bioorganic Chemistry, Kyoritsu College of Pharmacy, Minato-ku, Tokyo, Japan

    Y Nakazawa, N Manabe & M Mochizuki

  3. Harvard Institutes of Medicine, Harvard Medical School, Boston, MA, USA

    D G Tenen

  4. Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Chiba, Japan

    A Iwama

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  1. Y Nakazawa
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  2. M Suzuki
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  4. T Yamada
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Correspondence to T Oikawa.

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Nakazawa, Y., Suzuki, M., Manabe, N. et al. Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression. Oncogene 26, 3541–3550 (2007). https://doi.org/10.1038/sj.onc.1210140

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