title: Differential glucocorticoid enhancement of the cytokine-driven transcriptional activation of the human acute phase serum amyloid A genes, SAA1 and SAA2.
abstract: The human acute phase serum amyloid A (A-SAA) genes, SAA1 and SAA2, have a high degree of sequence identity that extends approximately 450 bp upstream of their transcription start sites. Each promoter contains analogously positioned functional binding sites for the transcription factors NF-kappaB and NF-IL6. In human HepG2 hepatoma cells transfected with SAA promoter luciferase reporter constructs, administration of IL-1 and IL-6, singly or in combination, induced SAA1 and SAA2 transcriptional readouts that were qualitatively indistinguishable. However, under induced conditions, the SAA2 promoter had a significant quantitative transcriptional advantage over the SAA1 promoter. The application of the synthetic glucocorticoid dexamethasone in the context of cytokine stimulation enhanced the transcriptional activity of the SAA1, but not the SAA2, promoter such that readout from the former became equivalent to that from the latter. A putative glucocorticoid response element (GRE) is present (between residues -208 and -194) only in the SAA1 gene; a similar sequence in the corresponding region of the SAA2 gene is disrupted by a nine-residue insertion. The SAA1 GRE was shown to be functionally active and the SAA2 disrupted GRE was shown to be functionally inactive in experiments using reporter constructs carrying SAA1 and SAA2 promoters that had been modified by site-specific mutagenesis. Quantitative analysis of transcript-specific RT-PCR products, derived from SAA1 and SAA2 mRNAs after treatment of HepG2 cells with cytokines in the presence or absence of dexamethasone, confirmed that the endogenous SAA1 gene has a cytokine-driven transcriptional disadvantage that is superseded by a marginal transcriptional advantage when glucocorticoids are present.
authors.affiliation: Department of Pharmacology and Center for Pharmacogenetics, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Caroline F
authors.initials: CF
authors.lastname: Thorn
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Whitehead
mesh_terms: D000209:Acute-Phase Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D001483:Base Sequence; D016207:Cytokines / Q000494:pharmacology / Q000502:physiology*; D003907:Dexamethasone / Q000494:pharmacology*; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000276:immunology; D011965:Receptors, Glucocorticoid / Q000096:biosynthesis / Q000502:physiology; D020218:Response Elements / Q000187:drug effects / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D000685:Serum Amyloid A Protein / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation / Q000187:drug effects* / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000209:Acute-Phase Proteins; D000276:Adjuvants, Immunologic; D016207:Cytokines; D011965:Receptors, Glucocorticoid; C083772:SAA2 protein, human; D000685:Serum Amyloid A Protein; D003907:Dexamethasone