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Characterization of the human CIDEA promoter in fat cells

International Journal of Obesity volume 32, pages 1380–1387 (2008)Cite this article

Abstract

Background:

Cell death-inducing DFFA (DNA fragmentation factor-α)-like effector A (CIDEA) is a protein that regulates lipolysis in human adipocytes through cross-talk involving tumor necrosis factor-α (TNF-α). TNF-α downregulates CIDEA mRNA although it is unclear whether this is mediated through transcriptional or post-transcriptional mechanisms. CIDEA has important metabolic effects in human fat cells and genetic variations in the human CIDEA gene have been correlated to the development of obesity. However, little is known about the factors regulating CIDEA expression in human adipocytes. We set out to describe the transcriptional control of human CIDEA.

Methods:

A 1.1-kb genomic fragment upstream of the transcriptional start site (TSS) of human CIDEA was cloned and deletion fragments were generated. Transcriptional activity of the promoter was analyzed by luciferase reporter assays in in vitro-differentiated human adipocytes. The effect of TNF-α was assessed in human adipocytes and murine 3T3-L1 cells transfected with deletion fragments of the CIDEA promoter. Protein–DNA interactions were analyzed by electrophoretic mobility shift assays (EMSA).

Results:

Basal transcriptional activity was found in a 97-bp region upstream of the TSS. We studied the effect of three common haplotypes in the promoter region but found no significant difference in transcriptional activity among them. Incubation of in vitro-differentiated human adipocytes as well as 3T3-L1 cells with TNF-α reduced the transcriptional activity of the human CIDEA promoter, demonstrating a direct effect on CIDEA transcription. EMSAs and mutational analysis indicated that this was mediated by a nuclear factor-κB (NF-κB) site at position −163/−151.

Conclusion:

We demonstrate that basal transcription of the human CIDEA gene is confined to the 97 first bases upstream of TSS and that TNF-α negatively regulates transcription of this gene, which at least in part involves NF-κB activation.

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Acknowledgements

We thank Gaby Åström for excellent technical assistance. This work was supported by grants from the Swedish Research Council, Swedish Heart and Lung Foundation, Swedish Diabetes Association, The memorial foundation of Sigurd and Elsa Golje, Storstockholms Diabetesförening, Torsten och Ragnar Söderbergs stiftelse Novo Nordisk Foundation, the European Union (HEPADIP, LSHM-CT-2005-018734), the Swedish Medical Association and the Swedish Diabetes Foundation.

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Authors and Affiliations

  1. Department of Medicine, Huddinge, Lipid Laboratory, Novum, Karolinska Institutet, Stockholm, Sweden

    A T Pettersson, J Laurencikiene, E A Nordström, B M Stenson, V van Harmelen, I Dahlman & M Rydén

  2. Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden

    C Murphy

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  1. A T Pettersson
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  2. J Laurencikiene
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  3. E A Nordström
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  4. B M Stenson
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  5. V van Harmelen
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  6. C Murphy
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  7. I Dahlman
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Correspondence to A T Pettersson.

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Pettersson, A., Laurencikiene, J., Nordström, E. et al. Characterization of the human CIDEA promoter in fat cells. Int J Obes 32, 1380–1387 (2008). https://doi.org/10.1038/ijo.2008.101

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