Competitive binding of STATs to receptor phospho-Tyr motifs accounts for altered cytokine responses

Wilmes, Stephan; Jeffrey, Polly-Anne; Martínez-Fábregas, Jonathan; Hafer, Maximillian; Fyfe, Paul K.; Pöhler, Elizabeth; Gaggero, Silvia; López‐García, Martín; Lythe, Grant; Taylor, Charles; Guerrier, Thomas; Launay, David; Mitra, Suman; Piehler, Jacob; Molina-Parı́s, Carmen; Moraga, Ignacio

doi:10.7554/elife.66014

Cited by 22 publications

(26 citation statements)

References 102 publications

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“…IL-27 signals through dimerization of the cognate receptor IL-27Rα and the shared receptor GP130 25 . Current models for the assembly of complexes mediated by IL-27 have been largely based on structural principles derived from the IL-12/IL-23 or IL-6 systems.…”

Section: Resultsmentioning

confidence: 99%

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Structural insights into the assembly and activation of IL-27 signalling complex

Jin

Fyfe

Gardner

et al. 2022

Preprint

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Interleukin 27 (IL-27) is a heterodimeric cytokine that elicits potent immuno-suppressive responses. Comprised of EBI3 and p28 subunits, IL-27 binds GP130 and IL-27Rα receptor chains to activate the JAK/STAT signalling cascade. However, how these receptors recognize IL-27 and form a complex capable of phosphorylating JAK proteins remains unclear. Here, we used cryo electron microscopy (cryoEM) to solve the structure of the IL-27 receptor recognition complex. Our data show how IL-27 serves as a bridge connecting IL-27Rα with GP130 to initiate signalling. While both receptors weakly bind the p28 component of the heterodimeric cytokine, EBI3 stabilizes the complex by binding a positively charged surface of IL-27Rα. We observe a large degree of flexibility in the rotation of D1 and the two CHR domains of GP130 contacting p28, which could contribute to GP130 binding degeneracy. We find that assembly of the IL-27 receptor recognition complex is distinct from both IL-12 and IL-6 cytokine families and provides a mechanistic blueprint for tuning IL-27 pleiotropic actions.

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Section: Resultsmentioning

confidence: 99%

“…To overcome challenges in cytokine stability, we engineered an IL-27 cytokine variant in which the two monomeric components (EBI3 and p28) were fused by a short linker 25 . The fusion cytokine was expressed in insect cells and purified together with Domains 1-3 (D1, D2, D3) of GP130 and the first two domains of IL-27Rα (Supplementary Fig.…”

Section: Resultsmentioning

confidence: 99%

Structural insights into the assembly and activation of IL-27 signalling complex

Jin

Fyfe

Gardner

et al. 2022

Preprint

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“…IL-27 signals through a receptor complex consisting of IL-27Rα (TCCR/WSX-1) and gp130 expressed predominantly on T cells and NK cells (Pflanz et al, 2002, 2004) (Figure 1A). Binding of IL-27 to its receptor subunits triggers the activation of receptor-associated Janus Kinase 1 (JAK1) and JAK2 leading to phosphorylation of STAT1 and STAT3 (Lucas et al, 2003; Owaki et al, 2008; Wilmes et al, 2021). Functionally, IL-27 signaling serves to constrain inflammation by antagonizing differentiation of pro-inflammatory Th17 cells (Stumhofer et al, 2006), stimulating T-bet expression in regulatory T cells (Tregs) (Hall et al, 2012), and inducing production of the anti-inflammatory cytokine IL-10 (Stumhofer et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Structure of the IL-27 quaternary receptor signaling complex

Caveney

Glassman

Jude

et al. 2022

Preprint

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Interleukin 27 (IL-27) is a heterodimeric cytokine that functions to constrain T cell-mediated inflammation and plays an important role in immune homeostasis. Binding of IL-27 to cell surface receptors IL-27Ra and gp130 results in activation of receptor-associated Janus Kinases and nuclear translocation of STAT1 and STAT3 transcription factors. Despite the emerging therapeutic importance of this cytokine axis in cancer and autoimmunity, a molecular blueprint of the IL-27 receptor signaling complex, and its relation to other gp130/IL-12 family cytokines, is currently unclear. We used cryogenic-electron microscopy (cryo-EM) to determine the quaternary structure of IL-27 (p28/Ebi3) bound to receptor subunits, IL-27Ra and gp130. The resulting 3.47 A resolution structure revealed a three-site assembly mechanism nucleated by the central p28 subunit of the cytokine. The overall topology and molecular details of this binding are reminiscent of IL-6 but distinct from related heterodimeric cytokines IL-12 and IL-23. These results indicate distinct receptor assembly mechanisms used by heterodimeric cytokines with important consequences for targeted agonism and antagonism of IL-27 signaling.

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“…In previous studies, combining experimental work with mathematical methods has been a successful approach to gain quantitative insights into Th cell dynamics and decision-making (9)(10)(11)(12)(13)(14)(15). Notably, it was found that although signal integration via cytokines is transient and stochastic (16,17), the resulting decisions regarding the generation of T cell phenotypes, including selective cytokine secretion, are remarkably stable even in quantitative terms at the single-cell level (10).…”

Section: Introductionmentioning

confidence: 99%

High-resolution kinetic gene expression analysis of T helper cell differentiation reveals a STAT-dependent, unique transcriptional program in Th1/2 hybrid cells

Burt

Peine

Borek

et al. 2022

Preprint

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Selective differentiation of CD4+ T helper (Th) cells into specialized subsets such as Th1 and Th2 cells is a key element of the adaptive immune system driving appropriate immune responses. Besides those canonical Th cell lineages, hybrid phenotypes such as Th1/2 cells arise in vivo, and their generation could be reproduced in vitro. While master-regulator transcription factors like T-bet for Th1 and GATA-3 for Th2 cells drive and maintain differentiation into the canonical lineages, the transcriptional architecture of hybrid phenotypes is less well understood. In particular, it has remained unclear whether a hybrid phenotype implies a mixture of the effects of several canonical lineages for each gene, or rather a bimodal behavior across genes. Th cell differentiation is a dynamic process in which the regulatory factors are modulated over time, but longitudinal studies of Th cell differentiation are sparse. Here, we present a dynamic transcriptome analysis following Th cell differentiation into Th1, Th2 and Th1/2 hybrid cells. We identified an early bifurcation point in gene expression programs, and we found that only a minority of ∼20% of Th cell-specific genes showed mixed effects from both Th1 and Th2 cells on Th1/2 hybrid cells. While most genes followed either Th1 or Th2 cell gene expression, another fraction of ∼20% of genes followed a Th1 and Th2 cell-independent transcriptional program under control of the transcription factors STAT1 and STAT4. Overall, our results emphasize the key role of high-resolution longitudinal data for the characterization of cellular phenotypes.

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Competitive binding of STATs to receptor phospho-Tyr motifs accounts for altered cytokine responses

Cited by 22 publications

References 102 publications

Structural insights into the assembly and activation of IL-27 signalling complex

Structural insights into the assembly and activation of IL-27 signalling complex

Structure of the IL-27 quaternary receptor signaling complex

High-resolution kinetic gene expression analysis of T helper cell differentiation reveals a STAT-dependent, unique transcriptional program in Th1/2 hybrid cells

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