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NF-Y regulates LIF-induced transcription of the signaling adaptor SKAP55R in myeloid cells

Leukemia volume 15, pages 1932–1940 (2001)Cite this 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 (M1 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.

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Acknowledgements

This work was supported by grants from the National Institutes of Health and a National Health and Medical Research Council CJ Martin Fellowship (DJC).

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Authors and Affiliations

  1. Genetics and Molecular Biology Branch, National Human Genome Research Institute, Bethesda, MD, USA

    DJ Curtis & DM Bodine

  2. Cancer and Haematology Division, Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia

    DJ Curtis & L Dougherty

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Curtis, D., Dougherty, L. & Bodine, D. NF-Y regulates LIF-induced transcription of the signaling adaptor SKAP55R in myeloid cells. Leukemia 15, 1932–1940 (2001). https://doi.org/10.1038/sj.leu.2402286

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