Characterization of the mouse interleukin-5 promoter in a mouse TH2 T cell clone.
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).
Stranick
K S
KS
Department of Allergy, Schering-Plough Research Institute, 2015 Galloping Hill Road, Kenilworth, New Jersey, 07033, USA.
Uss
A S
AS
Zambas
D N
DN
Egan
R W
RW
Billah
M M
MM
Umland
S P
SP
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D002999:Clone Cells; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D003850:Deoxyribonuclease I; D015848:Interleukin-5 / Q000235:genetics*; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D018418:Th2 Cells / Q000276:immunology*
D017931:DNA Primers; D004268:DNA-Binding Proteins; D015848:Interleukin-5; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D008156:Luciferases; D003850:Deoxyribonuclease I