NF-Y regulates LIF-induced transcription of the signaling adaptor SKAP55R in myeloid cells
- Author(s)
- Curtis, DJ; Dougherty, L; Bodine, DM;
- Details
- Publication Year 2001-12-01,Volume 15,Issue #12,Page 1932-1940
- Journal Title
- LEUKEMIA
- Publication Type
- Journal Article
- Abstract
- Previously, we have shown that interleukin-6 (IL-6) or leukemia inhibitory factor (LIF)-induced differentiation of the myeloid cell line M1 was associated with a rapid increase in the level of mRNA encoding the signaling adaptor protein, SKAP55R. Furthermore, enforced over-expression of SKAP55R in primary bone marrow cells inhibited colony growth. In this study, we have identified the cis-acting elements that control SKAP55R transcription in myeloid cells. The 980 bp genomic sequence upstream of the transcriptional start site was cloned into a GFP reporter vector for transient (293 cells) or stable (Ml cells) transfection assays. This region contained cis-acting elements necessary for transcriptional activity in unstimulated 293 cells (10-fold higher levels than the control vector) or unstimulated M1 cells (two-fold higher levels). Significant LIF-induced transcription was observed in M1 (3.4-fold induction, P < 0.001), but not 293 cells. Deletion reporter constructs defined a promoter region (-317/-137) essential for the transcriptional activity in M1 cells. This region contained a CCAAT element recently implicated in IL-6/LIF-induced transcriptional regulation of junB in M1 cells. Mutation of the CCAAT element (-250/-246) significantly reduced both basal and LIF-induced transcription (P < 0.01). Electrophoretic mobility shift assays demonstrated that NF-Y bound to the CCAAT element of both SKAP55R and junB. These results suggest NF-Y binding may be a common mechanism of IL-6/LIF-regulated transcription in myeloid cells.
- Publisher
- NATURE PUBLISHING GROUP
- Keywords
- PHASE RESPONSE FACTOR; DNA-BINDING PROTEINS; MOLECULAR-CLONING; NUCLEAR FACTOR; GROWTH ARREST; GENE PROMOTER; MYD GENES; KAPPA-B; INTERLEUKIN-6; GP130
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Creation Date: 2001-12-01 12:00:00