The Journal of Immunology, Vol 158, Issue 2 748-755, Copyright © 1997 by American Association of Immunologists

ARTICLES

Intracellular IL-1 receptor antagonist promoter: cell type-specific and inducible regulatory regions

JK Jenkins, RF Drong, ME Shuck, MJ Bienkowski, JL Slightom, WP Arend and MF Smith Jr
Division of Rheumatology, Department of Medicine, University of Colorado Health Sciences Center, Denver 80262, USA.

The objective of these studies was to examine the molecular mechanisms involved in transcriptional regulation of the gene for the intracellular structural variant of the IL-1 receptor antagonist (icIL- 1Ra) molecule. By reverse transcription-PCR analysis, constitutive expression of endogenous icIL-1Ra mRNA was observed in the epithelial cell lines A431 and HT-29, but not in the macrophage cell lines RAW 264.7 and U937, or in the lymphocyte cell lines Raji and Jurkat. However, icIL-1Ra mRNA expression was observed in response to stimulation with LPS in RAW 264.7 cells and to PMA and LPS in U937 cells. To examine the mechanisms of transcriptional regulation, 4.5 kb of the 5' flanking sequence was isolated from the human icIL-1Ra gene, sequenced, cloned into a luciferase expression vector (pIC4525.Luc), and examined in transfection studies. The pIC4525.Luc construct exhibited a pattern of expression in epithelial and macrophage cell lines similar to that of the endogenous icIL-1Ra gene. To obtain a generalized map of cell type-specific and inducible cis-acting DNA elements, nested 5' deletional mutants of the icIL-1Ra promoter were constructed. Results from transfection studies with these icIL-1Ra promoter/luciferase fusion constructs indicated that constitutive expression in epithelial cells was under the control of three positively acting regions located between bases -4525 to -1438, -288 to -156, and -156 to -49. In contrast, basal expression of pIC4525.Luc in transfected but unstimulated RAW 264.7 cells was under the control of a weak inhibitory region located between bases -4525 to -1438 and a strong positive element between -156 and -49. LPS induction of icIL-1Ra transcription in RAW 264.7 cells was regulated by strong positively acting DNA regions between bases -1438 to -909 and -156 to -49. In summary, the proximal region of the icIL-1Ra promoter, between bases - 156 to -49, contains positive cis-acting elements that are needed for expression in both epithelial and monocyte cell lines. However, our results indicate that the ability of this proximal promoter region to control expression is strongly influenced, both positively and negatively, by other upstream cis-acting elements in a cell type- specific manner.

This article has been cited by other articles:

				A. P. Reiner, M. M. Wurfel, L. A. Lange, C. S. Carlson, A. S. Nord, C. L. Carty, M. J. Rieder, C. Desmarais, N. S. Jenny, C. Iribarren, et al. Polymorphisms of the IL1-Receptor Antagonist Gene (IL1RN) Are Associated With Multiple Markers of Systemic Inflammation Arterioscler. Thromb. Vasc. Biol., July 1, 2008; 28(7): 1407 - 1412. [Abstract] [Full Text] [PDF]

				J. Kong, S. A. Grando, and Y. C. Li Regulation of IL-1 Family Cytokines IL-1{alpha}, IL-1 Receptor Antagonist, and IL-18 by 1,25-Dihydroxyvitamin D3 in Primary Keratinocytes J. Immunol., March 15, 2006; 176(6): 3780 - 3787. [Abstract] [Full Text] [PDF]

				F. Merhi-Soussi, B. R. Kwak, D. Magne, C. Chadjichristos, M. Berti, G. Pelli, R. W. James, F. Mach, and C. Gabay Interleukin-1 plays a major role in vascular inflammation and atherosclerosis in male apolipoprotein E-knockout mice Cardiovasc Res, June 1, 2005; 66(3): 583 - 593. [Abstract] [Full Text] [PDF]

				E. La and S. M. Fischer Transcriptional Regulation of Intracellular IL-1 Receptor Antagonist Gene by IL-1{{alpha}} in Primary Mouse Keratinocytes J. Immunol., May 15, 2001; 166(10): 6149 - 6155. [Abstract] [Full Text] [PDF]

				C. Gabay, M. G. Dreyer, N. Pellegrinelli, R. Chicheportiche, and C. A. Meier Leptin Directly Induces the Secretion of Interleukin 1 Receptor Antagonist in Human Monocytes J. Clin. Endocrinol. Metab., February 1, 2001; 86(2): 783 - 791. [Abstract] [Full Text]

				G. C. Higgins, Y. Wu, and A. E. Postlethwaite Intracellular IL-1 Receptor Antagonist Is Elevated in Human Dermal Fibroblasts That Overexpress Intracellular Precursor IL-1{alpha} J. Immunol., October 1, 1999; 163(7): 3969 - 3975. [Abstract] [Full Text] [PDF]

				S. Lemay, T. V. Lebedeva, and A. K. Singh Inhibition of Cytokine Gene Expression by Sodium Salicylate in a Macrophage Cell Line through an NF-kappa B-Independent Mechanism Clin. Vaccine Immunol., July 1, 1999; 6(4): 567 - 572. [Abstract] [Full Text] [PDF]

				B. P. Haines, R. B. Voyle, T. A. Pelton, R. Forrest, and P. D. Rathjen Complex Conserved Organization of the Mammalian Leukemia Inhibitory Factor Gene: Regulated Expression of Intracellular and Extracellular Cytokines J. Immunol., April 15, 1999; 162(8): 4637 - 4646. [Abstract] [Full Text] [PDF]

				M. F. Smith Jr., V. S. Carl, T. Lodie, and M. J. Fenton Secretory Interleukin-1 Receptor Antagonist Gene Expression Requires both a PU.1 and a Novel Composite NF-kappa B/PU.1/ GA-binding Protein Binding Site J. Biol. Chem., September 11, 1998; 273(37): 24272 - 24279. [Abstract] [Full Text] [PDF]

				M. Malyak, J. M. Guthridge, K. R. Hance, S. K. Dower, J. H. Freed, and W. P. Arend Characterization of a Low Molecular Weight Isoform of IL-1 Receptor Antagonist J. Immunol., August 15, 1998; 161(4): 1997 - 2003. [Abstract] [Full Text] [PDF]