Analysis of cis-acting sequences and trans-acting factors regulating the interleukin-3 response element of the DUB-1 gene.
The murine DUB-1 gene is a hematopoietic-specific, immediate-early gene that encodes a growth-regulatory deubiquitinating enzyme. DUB-1 contains an IL-3-inducible enhancer element that is activated in a JAK2-dependent, STAT5-independent manner. In this study, we have further characterized this novel IL-3 response element. Transcriptional reporter assays in Ba/F3 cells revealed that two AP-1 sites, a GATA motif, and an Ets site are required for induction of DUB-1 enhancer activity. Gel shift assays indicated that IL-3 activates the binding of an AP-1 complex containing JunD to the AP-1 sites and the binding of another protein complex to the Ets motif. The latter complex was not detectable in Ba/F3 cells stably transfected with a dominant-negative mutant of JAK2. As previously shown, these cells do not express DUB-1 mRNA or protein. Furthermore, we demonstrated that GATA-1 constitutively binds to the DUB-1 enhancer element. The involvement of GATA-1 may be important for the hematopoietic-restricted expression pattern of DUB-1. This combination of inducible and constitutive elements of the DUB-1 enhancer appears to account for the unique STAT-independent expression characteristics of DUB-1.
Jaster
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Division of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA.
Baek
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D'Andrea
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D000818:Animals; D002478:Cells, Cultured; D016384:Consensus Sequence; D010450:Endopeptidases; D004742:Enhancer Elements, Genetic / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D017874:Immediate-Early Proteins / Q000235:genetics*; D007377:Interleukin-3 / Q000494:pharmacology*; D053614:Janus Kinase 2; D051379:Mice; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000494:pharmacology; D011518:Proto-Oncogene Proteins; D020218:Response Elements / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D015534:Trans-Activators
D017874:Immediate-Early Proteins; D007377:Interleukin-3; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D011505:Protein-Tyrosine Kinases; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2; C099271:Dub1 protein, mouse; D010450:Endopeptidases