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Oncogenic Ras upregulates NADPH oxidase 1 gene expression through MEK-ERK-dependent phosphorylation of GATA-6

Oncogene volume 27, pages 4921–4932 (2008)Cite this article

Abstract

Ras oncogene upregulates the expression of nicotinamide adenine dinucleotide phosphate oxidase (Nox) 1 via the Raf/MEK/ERK pathway, leading to the elevated production of reactive oxygen species that is essential for maintenance of Ras-transformation phenotypes. However, the precise transcriptional control mechanism underlying Ras-induced Nox1 expression remains to be elucidated. Here we demonstrated that via the MEK/ERK pathway, Ras signaling enhances the activity of the functional Nox1 promoter (nt −321 to −1) in colon cancer CaCo-2 cells and thereby induces the formation of the specific protein–DNA complexes in the two GATA-binding site-containing regions (nt −161 to −136 and −125 to −100). Supershift assays with GATA antibodies, protein analyses and chromatin immunoprecipitation revealed that GATA-6 is a component of the specific protein–DNA complexes at the Nox1 promoter. GATA-6 was able to trans-activate the Nox1 promoter but not a promoter in which the GATA-binding sites are mutated. Moreover, GATA-6 was phosphorylated at serine residues by MEK-activated ERK, which increased GATA-6 DNA binding, correlating with suppression of the Nox1 promoter activity by an MEK inhibitor PD98059. Finally, the site-directed mutation of the consensus ERK phosphorylation site (PYS120P to PYA120P) of GATA-6 abolished its trans-activation activity, suppressing of the growth of CaCo-2 cells. On the basis of these results, we propose that oncogenic Ras signaling upregulates the transcription of Nox1 through MEK-ERK-dependent phosphorylation of GATA-6.

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Acknowledgements

We thank Dr J Downward, Dr E Nishida and Dr M Maeda for plasmid DNAs. This study was supported by Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology of Japan (TK).

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, Shinshu University School of Medicine, Nagano, Japan

    Y Adachi, Y Shibai, J Mitsushita, W H Shang & T Kamata

  2. Biomedical Research Institute, Kureha Chemical Industry Co. Ltd, Tokyo, Japan

    K Hirose

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  1. Y Adachi
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Correspondence to T Kamata.

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Adachi, Y., Shibai, Y., Mitsushita, J. et al. Oncogenic Ras upregulates NADPH oxidase 1 gene expression through MEK-ERK-dependent phosphorylation of GATA-6. Oncogene 27, 4921–4932 (2008). https://doi.org/10.1038/onc.2008.133

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