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SMAD3 inhibits SF-1-dependent activation of the CYP17 promoter in H295R cells

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Abstract

Cytochrome P450c17, encoded by the CYP17 gene, is a component of 17α-hydroxylase/17,20 lyase which catalyses 17α-hydroxylation of pregnenolone or progesterone, required for glucocorticosteroid and androgen synthesis. It has been reported that transforming growth factor β (TGF-β) decreases both basal and cAMP-stimulated levels of CYP17 mRNA, but the mechanism of TGF-β action on CYP17 expression remains unknown. We investigated an inhibitory effect of TGF-β on CYP17 expression in H295R cells using constructs containing the CYP17 promoter region fused with the luciferase gene. In the H295R cells, TGF-β decreased endogenous SF-1 level and inhibited activity of the 300 bp fragment of CYP17 promoter, which was stimulated by coexpression of SF-1. Overexpression of SMAD3 caused an inhibition of SF-1-stimulated CYP17 promoter activity, whereas overexpression of SMAD7 was ineffective. In conclusion, our results suggest that the inhibitory action of TGF-β on CYP17 transcription involve at least two mechanisms: SMAD3 dependent inactivation of CYP17 promoter activity and repression of SF-1 expression.

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Abbreviations

SF-1:

Steroidogenic factor 1 (NR5A1, Ad4BP)

TGF-β:

Transforming growth factor β

TβRII:

Type II TGF-β receptor

StAR:

Steroidogenic acute regulatory protein

SBE:

SMAD binding element

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Acknowledgements

Supported by a grant No P04A 03127 from The State Committee for Scientific Research (KBN). N.D-H. and T.P.L. are recipients of scholarships from the Postgraduate School of Molecular Medicine affiliated with the Medical University of Warsaw. We are grateful to graduate student Kamila Jasiak for the help in coimmunoprecipitation and Western blotting and to Mr. Jason Banks for expert editorial assistance.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical University, 6 Swiecickiego St., Poznan, 60-781, Poland

    Natalia Derebecka-Holysz, Tomasz P. Lehmann, Marcin Holysz & Wieslaw H. Trzeciak

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  1. Natalia Derebecka-Holysz
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  2. Tomasz P. Lehmann
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  3. Marcin Holysz
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  4. Wieslaw H. Trzeciak
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Correspondence to Tomasz P. Lehmann.

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Derebecka-Holysz, N., Lehmann, T.P., Holysz, M. et al. SMAD3 inhibits SF-1-dependent activation of the CYP17 promoter in H295R cells. Mol Cell Biochem 307, 65–71 (2008). https://doi.org/10.1007/s11010-007-9585-4

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  • DOI: https://doi.org/10.1007/s11010-007-9585-4

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