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

Yosuke Nakazawa, M. Suzuki, N. Manabe, T. Yamada, F. Kihara-Negishi, T. Sakurai, D. G. Tenen, A. Iwama, M. Mochizuki, T. Oikawa

Research output: Contribution to journalArticle

27 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)3541-3550
Number of pages10
JournalOncogene
Volume26
Issue number24
DOIs
Publication statusPublished - 2007 May 24
Externally publishedYes

Fingerprint

Intercellular Signaling Peptides and Proteins
Transcription Factors
Gene Expression
Transcriptional Activation
Binding Sites
Chromatin Immunoprecipitation
Oncogene Proteins
Zinc Fingers
Site-Directed Mutagenesis
Glutathione Transferase
Luciferases
Immunoprecipitation
Small Interfering RNA
Cell Differentiation
Growth

Keywords

  • Bax
  • Cross-talk
  • ETS1
  • GFI1
  • Transcription

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Nakazawa, Y., Suzuki, M., Manabe, N., Yamada, T., Kihara-Negishi, F., Sakurai, T., ... Oikawa, T. (2007). Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression. Oncogene, 26(24), 3541-3550. https://doi.org/10.1038/sj.onc.1210140

Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression. / Nakazawa, Yosuke; Suzuki, M.; Manabe, N.; Yamada, T.; Kihara-Negishi, F.; Sakurai, T.; Tenen, D. G.; Iwama, A.; Mochizuki, M.; Oikawa, T.

In: Oncogene, Vol. 26, No. 24, 24.05.2007, p. 3541-3550.

Research output: Contribution to journalArticle

Nakazawa, Y, Suzuki, M, Manabe, N, Yamada, T, Kihara-Negishi, F, Sakurai, T, Tenen, DG, Iwama, A, Mochizuki, M & Oikawa, T 2007, 'Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression', Oncogene, vol. 26, no. 24, pp. 3541-3550. https://doi.org/10.1038/sj.onc.1210140
Nakazawa, Yosuke ; Suzuki, M. ; Manabe, N. ; Yamada, T. ; Kihara-Negishi, F. ; Sakurai, T. ; Tenen, D. G. ; Iwama, A. ; Mochizuki, M. ; Oikawa, T. / Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression. In: Oncogene. 2007 ; Vol. 26, No. 24. pp. 3541-3550.
@article{489ce6fa8d684c97bba3b451767c699d,
title = "Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression",
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.",
keywords = "Bax, Cross-talk, ETS1, GFI1, Transcription",
author = "Yosuke Nakazawa and M. Suzuki and N. Manabe and T. Yamada and F. Kihara-Negishi and T. Sakurai and Tenen, {D. G.} and A. Iwama and M. Mochizuki and T. Oikawa",
year = "2007",
month = "5",
day = "24",
doi = "10.1038/sj.onc.1210140",
language = "English",
volume = "26",
pages = "3541--3550",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "24",

}

TY - JOUR

T1 - Cooperative interaction between ETS1 and GFI1 transcription factors in the repression of Bax gene expression

AU - Nakazawa, Yosuke

AU - Suzuki, M.

AU - Manabe, N.

AU - Yamada, T.

AU - Kihara-Negishi, F.

AU - Sakurai, T.

AU - Tenen, D. G.

AU - Iwama, A.

AU - Mochizuki, M.

AU - Oikawa, T.

PY - 2007/5/24

Y1 - 2007/5/24

N2 - 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.

AB - 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.

KW - Bax

KW - Cross-talk

KW - ETS1

KW - GFI1

KW - Transcription

UR - http://www.scopus.com/inward/record.url?scp=34249295972&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34249295972&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1210140

DO - 10.1038/sj.onc.1210140

M3 - Article

VL - 26

SP - 3541

EP - 3550

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 24

ER -