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Characterization of the human CIDEA promoter in fat cells.
Int J Obes (Lond). 2008 Sep; 32(9):1380-7.IJ

Abstract

BACKGROUND

Cell death-inducing DFFA (DNA fragmentation factor-alpha)-like effector A (CIDEA) is a protein that regulates lipolysis in human adipocytes through cross-talk involving tumor necrosis factor-alpha (TNF-alpha). TNF-alpha 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-alpha 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-alpha 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-kappaB (NF-kappaB) 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-alpha negatively regulates transcription of this gene, which at least in part involves NF-kappaB activation.

Authors+Show Affiliations

Department of Medicine, Huddinge, Lipid Laboratory, Novum, Karolinska Institutet, Stockholm, Sweden. amanda.pettersson@ki.seNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18607384

Citation

Pettersson, A T., et al. "Characterization of the Human CIDEA Promoter in Fat Cells." International Journal of Obesity (2005), vol. 32, no. 9, 2008, pp. 1380-7.
Pettersson AT, Laurencikiene J, Nordström EA, et al. Characterization of the human CIDEA promoter in fat cells. Int J Obes (Lond). 2008;32(9):1380-7.
Pettersson, A. T., Laurencikiene, J., Nordström, E. A., Stenson, B. M., van Harmelen, V., Murphy, C., Dahlman, I., & Rydén, M. (2008). Characterization of the human CIDEA promoter in fat cells. International Journal of Obesity (2005), 32(9), 1380-7. https://doi.org/10.1038/ijo.2008.101
Pettersson AT, et al. Characterization of the Human CIDEA Promoter in Fat Cells. Int J Obes (Lond). 2008;32(9):1380-7. PubMed PMID: 18607384.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of the human CIDEA promoter in fat cells. AU - Pettersson,A T, AU - Laurencikiene,J, AU - Nordström,E A, AU - Stenson,B M, AU - van Harmelen,V, AU - Murphy,C, AU - Dahlman,I, AU - Rydén,M, Y1 - 2008/07/08/ PY - 2008/7/9/pubmed PY - 2009/3/28/medline PY - 2008/7/9/entrez SP - 1380 EP - 7 JF - International journal of obesity (2005) JO - Int J Obes (Lond) VL - 32 IS - 9 N2 - BACKGROUND: Cell death-inducing DFFA (DNA fragmentation factor-alpha)-like effector A (CIDEA) is a protein that regulates lipolysis in human adipocytes through cross-talk involving tumor necrosis factor-alpha (TNF-alpha). TNF-alpha 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-alpha 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-alpha 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-kappaB (NF-kappaB) 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-alpha negatively regulates transcription of this gene, which at least in part involves NF-kappaB activation. SN - 1476-5497 UR - https://www.unboundmedicine.com/medline/citation/18607384/Characterization_of_the_human_CIDEA_promoter_in_fat_cells_ L2 - https://doi.org/10.1038/ijo.2008.101 DB - PRIME DP - Unbound Medicine ER -