Binding of Octamer Factors to a Novel 3′-Positive Regulatory Element in the Mouse Interleukin-5 Gene

The development of eosinophilia is regulated by interleukin (IL)-5. The biological specificity of eosinophilia suggests a tight and independent regulation of IL-5 expression. A number of regulatory regions in the 5'-end of the IL-5 gene have been described; many of them are involved in the regulation of other genes, and it is not clear how the specific expression of IL-5 is regulated. In this study, we report the finding of a novel 3'-regulatory element. Data base analysis of a 2-kilobase fragment of the 3'-end of the mouse IL-5 gene revealed the presence of a 40-base pair-long repetitive sequence that consists of four direct repeats of ATGAATGA distributed in a symmetrical manner. This sequence, named mouse downstream regulatory element-1 (mDRE1), was shown to be protected in DNase I footprinting assays in vitro. Electrophoretic mobility shift assays using specific antibodies identified the transcription factors Oct-1 and Oct-2 as responsible for the formation of the specific complexes with mDRE1 and nuclear extracts from both EL4 and primary T-cells. Competition electrophoretic mobility shift assays with oligonucleotides containing different numbers of ATGAATGA repeats showed that Oct-1 and Oct-2 bind to different motifs in the mDRE1 sequence. Deletion of mDRE1 from a 9.5-kilobase IL-5 gene construct significantly decreased the expression of the luciferase reporter gene, suggesting that it plays a positive role in the expression of the IL-5 gene.