Comparison of two promoters controlling expression of secreted or intracellular IL-1 receptor antagonist.

Abstract

Abstract IL-1R antagonist can be expressed as a secreted glycoprotein (sIL-1Ra) or as an intracellular nonglycosylated form (icIL-1Ra). The two isoforms only differ in their N-terminal sequences and are thought to originate, by selection of alternative first exons, from a single gene (IL-1RN). We have previously described a clone that encodes the exons and promoter for sIL-1Ra. In this work, we describe and characterize a clone that contains the further upstream, alternative first exon used to generate icIL-1Ra. Sequences upstream to this alternative exon are demonstrated to have promoter activity in the human colon epithelial cell line HT29, whereas the sIL-1Ra promoter has undetectable activity. Both the icIL-1Ra and sIL-1Ra promoters were active when transfected into THP-1 cells. Neither promoter was responsive to PMA or LPS when introduced into HT29 or THP-1 cells. Activity of the endogenous icIL-1Ra and sIL-1Ra promoters was also investigated. In agreement with the transfected icIL-1Ra promoter, the endogenous icIL-1Ra gene is active in HT29 cells, though induced by PMA. In contrast to the transfected icIL-1Ra promoter, the endogenous icIL-1Ra gene is inactive in THP-1 cells and induced by PMA. Our data complete the structure of the human IL-1RN gene, demonstrating that the two forms of IL-1Ra are produced from the same gene by use of alternative first exons. We also show that there are two distinct promoters that control IL-1Ra expression, that lie upstream to each alternative first exon.