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Apoptosis

Characterization of a novel negative regulatory element in the human interleukin 4 promoter

Leukemia volume 14, pages 629–635 (2000)Cite this article

Abstract

Interleukin 4 (IL-4) is a multifunctional cytokine that plays an important role in hematopoiesis, tumor cell growth, and cellular immune responses. Expression of the IL-4 gene is tightly controlled at the level of gene transcription, and many positive regulatory cis-elements have been identified in the proximal IL-4 promoter region. Relatively little is known about factors that downregulate IL-4 transcription. We performed a detailed deletional analysis of the proximal human IL-4 promoter and studied reporter gene activity in transiently transfected Jurkat T lymphoblasts. In this report, we characterize a novel negative regulatory element (termed P2 NRE) that is adjacent to a binding site for nuclear factor of activated T cells. Mutation of P2 NRE significantly enhanced the activity of a 175 base pair IL-4 promoter construct in transiently transfected Jurkat T lymphoblasts. Using nuclear extracts from Jurkat cells, we identify a candidate factor (termed Rep-1) that binds uniquely to the P2 NRE in DNA-binding assays. Rep-1 is not related to other factors previously shown to interact with the IL-4 promoter, and by UV cross-linking and SDS-PAGE analysis, we found that it migrates with a molecular mass of approximately 150 kDa. Characterizing the molecular mechanisms responsible for downregulating the IL-4 promoter should enhance our understanding of IL-4-gene dysregulation in disease states.

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Acknowledgements

We thank Dr Anjana Rao and Dr Gerald Crabtree for the generous gifts of reagents. Cindy Chang and Mary Brummet provided invaluable assistance with the UV cross-linking experiments. This work was supported by grants from the National Institutes of Health, the American Lung Association, and the Johns Hopkins University School of Medicine Clinician Scientist Program.

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Author notes

  1. S Ono

    Present address: Laboratory of Molecular Immunology, The Schepens Eye Research Institute, Harvard Medical School, Boston, MA 02114, USA

Authors and Affiliations

  1. Division of Pulmonary and Critical Care Medicine, Johns Hopkins University Asthma and Allergy Center, Baltimore, MD, USA

    S Georas, J Cumberland, T Burke & E Park

  2. Division of Allergy and Clinical Immunology, Johns Hopkins University Asthma and Allergy Center, Baltimore, MD, USA

    S Georas, S Ono & V Casolaro

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Georas, S., Cumberland, J., Burke, T. et al. Characterization of a novel negative regulatory element in the human interleukin 4 promoter. Leukemia 14, 629–635 (2000). https://doi.org/10.1038/sj.leu.2401712

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