Characterization of the Mouse Interleukin-5 Promoter in a Mouse TH2 T Cell Clone

• Published in: Biochemical and biophysical research communications Vol. 252; no. 1; pp. 56 - 62
• Main Authors: Stranick, Kimberly S., Uss, Annette Schettino, Zambas, Demetris N., Egan, Robert W., Billah, M.Motasim, Umland, Shelby P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 09.11.1998
• Subjects: AIDS/HIV; Animals; Base Sequence; Binding Sites; Cell Nucleus - metabolism; Clone Cells; Deoxyribonuclease I; DNA Primers; DNA-Binding Proteins - metabolism; Interleukin-5 - genetics; Luciferases - biosynthesis; Mice; Molecular Sequence Data; Polymerase Chain Reaction; Promoter Regions, Genetic; Receptor-CD3 Complex, Antigen, T-Cell - immunology; Recombinant Fusion Proteins - biosynthesis; Th2 Cells - immunology
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1998.9594

The proximal mouse IL-5 promoter was examined using a mouse TH2 clone stimulated through the T cell receptor using anti-CD3 monoclonal antibody. DNase I protection defined four protein binding regions [IL-5RE-A, -69/-45; -B, (-90/-76); -C, (-154/-130); and -D (-176/-157)]. Stimulation-dependent binding, which was seen in the IL-5RE-B, -D regions and the 5′ end of tIL-5RE-A, did not require new protein synthesis inhibitor during cell activation. EMSA using probes targeted to the IL-5RE-B, -C, -D regions demonstrated the multimeric nature of the bound proteins. By transfection analysis using a series of truncated IL-5 promoter-luciferase constructs, IL-5RE-C and -D contributed little to constitutive or inducible activity. The CLE0 site in the IL-5RE-A region contributed to full transcriptional activity but was not sufficient to mediate full activity. Full stimulation-dependent activity required the IL-5RE-B region and/or the GATA site (-70/-60).