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IL-6 signaling via the STAT3/SOCS3 pathway: Functional Analysis of the Conserved STAT3 N-domain

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Abstract

The conserved N-domain of the STAT proteins has been implicated in several activities crucial to cytokine signaling including receptor recruitment and STAT activation, cooperative DNA binding and STAT-dependent gene expression. We evaluated the role of the STAT3 N-domain in the IL-6 signal transduction pathway leading to Socs3 gene expression, an essential mechanism that controls the quality and magnitude of IL-6-dependent transcriptional responses. Based on the model for STAT N-domain function in cooperative gene expression and the presence of tandem STAT binding motifs in the murine Socs3 promoter, we anticipated that stabilizing interactions between adjacent STAT3 dimers via N-domain sequences might be essential for Socs3 gene expression. This was underscored by the tight conservation in the location and sequence of the tandem STAT binding sites between the murine and human Socs3 promoters. Using reconstitution into Stat3−/− mouse embryonic fibroblasts (Stat3−/− MEFs), we find that a STAT3 N-domain deletion mutant (Δ 133STAT3) is activated by tyrosine phosphorylation in response to IL-6 and then undergoes dephosphorylation with kinetics similar to full-length STAT3. These results highlight important differences compared to other STATs where the N-domain has been shown to mediate activation (STAT4) or dephosphorylation (STAT1). STAT3 binds predominantly to a single STAT consensus site in the Socs3 promoter, despite the presence of an adjacent STAT motif. Significantly, Δ 133STAT3 stimulates expression of the endogenous Socs3 gene in Stat3−/− MEFs upon IL-6 treatment with an activity similar to reconstituted STAT3, demonstrating that the N-domain is dispensable for Socs3 gene expression. We propose that the Socs3 gene in its chromosomal context is activated by the IL-6/STAT3 pathway independent of STAT3 N-domain sequences.

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Authors and Affiliations

  1. Department of Immunology, The University of Texas MD Anderson Cancer Center, P.O. Box 301402, Unit 902, Houston, Texas, 77030-1903, USA

    Ling Zhang, Donna B. Badgwell, Jack J. Bevers III & Stephanie S. Watowich

  2. The Graduate School of Biomedical Sciences, University of Texas, Houston, Texas, 77030, USA

    Donna B. Badgwell, Jack J. Bevers III & Stephanie S. Watowich

  3. New York University School of Medicine, New York, NY, 10016, USA

    Karni Schlessinger & David E. Levy

  4. MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London, WC1E 6BT, USA

    Karni Schlessinger

  5. Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, TN, 38105, USA

    Peter J. Murray

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  1. Ling Zhang
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  2. Donna B. Badgwell
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Correspondence to Stephanie S. Watowich.

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Zhang, L., Badgwell, D.B., Bevers, J.J. et al. IL-6 signaling via the STAT3/SOCS3 pathway: Functional Analysis of the Conserved STAT3 N-domain. Mol Cell Biochem 288, 179–189 (2006). https://doi.org/10.1007/s11010-006-9137-3

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