Kinetics of Cytokine Receptor Trafficking Determine Signaling and Functional Selectivity

Overview

Journal Elife

Specialty Biology

Date 2019 Nov 28

PMID 31774398

Citations 24

Authors

Jonathan Martinez-Fabregas

Stephan Wilmes

Luopin Wang

Maximillian Hafer

Elizabeth Pohler

Juliane Lokau

Christoph Garbers

Adeline Cozzani

Paul K Fyfe

Jacob Piehler

Majid Kazemian

Suman Mitra

Ignacio Moraga

Affiliations

Soon will be listed here.

Abstract

Cytokines activate signaling via assembly of cell surface receptors, but it is unclear whether modulation of cytokine-receptor binding parameters can modify biological outcomes. We have engineered IL-6 variants with different affinities to gp130 to investigate how cytokine receptor binding dwell-times influence functional selectivity. Engineered IL-6 variants showed a range of signaling amplitudes and induced biased signaling, with changes in receptor binding dwell-times affecting more profoundly STAT1 than STAT3 phosphorylation. We show that this differential signaling arises from defective translocation of ligand-gp130 complexes to the endosomal compartment and competitive STAT1/STAT3 binding to phospho-tyrosines in gp130, and results in unique patterns of STAT3 binding to chromatin. This leads to a graded gene expression response and differences in ex vivo differentiation of Th17, Th1 and Treg cells. These results provide a molecular understanding of signaling biased by cytokine receptors, and demonstrate that manipulation of signaling thresholds is a useful strategy to decouple cytokine functional pleiotropy.

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References

Martinez-Fabregas J, Wilmes S, Wang L, Hafer M, Pohler E, Lokau J . Kinetics of cytokine receptor trafficking determine signaling and functional selectivity. Elife. 2019; 8. PMC: 6914340. DOI: 10.7554/eLife.49314. View

Wiederkehr-Adam M, Ernst P, Muller K, Bieck E, Gombert F, Ottl J . Characterization of phosphopeptide motifs specific for the Src homology 2 domains of signal transducer and activator of transcription 1 (STAT1) and STAT3. J Biol Chem. 2003; 278(18):16117-28. DOI: 10.1074/jbc.M300261200. View

You C, Marquez-Lago T, Richter C, Wilmes S, Moraga I, Garcia K . Receptor dimer stabilization by hierarchical plasma membrane microcompartments regulates cytokine signaling. Sci Adv. 2016; 2(12):e1600452. PMC: 5135388. DOI: 10.1126/sciadv.1600452. View

Lemmon M, Schlessinger J . Cell signaling by receptor tyrosine kinases. Cell. 2010; 141(7):1117-34. PMC: 2914105. DOI: 10.1016/j.cell.2010.06.011. View

Horvath C, Wen Z, Darnell Jr J . A STAT protein domain that determines DNA sequence recognition suggests a novel DNA-binding domain. Genes Dev. 1995; 9(8):984-94. DOI: 10.1101/gad.9.8.984. View

Marchetti M, Monier M, Fradagrada A, Mitchell K, Baychelier F, Eid P . Stat-mediated signaling induced by type I and type II interferons (IFNs) is differentially controlled through lipid microdomain association and clathrin-dependent endocytosis of IFN receptors. Mol Biol Cell. 2006; 17(7):2896-909. PMC: 1483027. DOI: 10.1091/mbc.e06-01-0076. View

Fallon E, Lauffenburger D . Computational model for effects of ligand/receptor binding properties on interleukin-2 trafficking dynamics and T cell proliferation response. Biotechnol Prog. 2000; 16(5):905-16. DOI: 10.1021/bp000097t. View

Moraga I, Richter D, Wilmes S, Winkelmann H, Jude K, Thomas C . Instructive roles for cytokine-receptor binding parameters in determining signaling and functional potency. Sci Signal. 2015; 8(402):ra114. PMC: 5568809. DOI: 10.1126/scisignal.aab2677. View

Huang W, Yan Q, Lin W, Chung J, Hansen S, Christensen S . Phosphotyrosine-mediated LAT assembly on membranes drives kinetic bifurcation in recruitment dynamics of the Ras activator SOS. Proc Natl Acad Sci U S A. 2016; 113(29):8218-23. PMC: 4961118. DOI: 10.1073/pnas.1602602113. View

10.

Boulanger M, Chow D, Brevnova E, Martick M, Sandford G, Nicholas J . Molecular mechanisms for viral mimicry of a human cytokine: activation of gp130 by HHV-8 interleukin-6. J Mol Biol. 2003; 335(2):641-54. DOI: 10.1016/j.jmb.2003.10.070. View

11.

Wilson E, Wille-Reece U, Dzierszinski F, Hunter C . A critical role for IL-10 in limiting inflammation during toxoplasmic encephalitis. J Neuroimmunol. 2005; 165(1-2):63-74. DOI: 10.1016/j.jneuroim.2005.04.018. View

12.

HAMERS-CASTERMAN C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa E . Naturally occurring antibodies devoid of light chains. Nature. 1993; 363(6428):446-8. DOI: 10.1038/363446a0. View

13.

Grotzinger J, Kurapkat G, Wollmer A, Kalai M, Rose-John S . The family of the IL-6-type cytokines: specificity and promiscuity of the receptor complexes. Proteins. 1997; 27(1):96-109. View

14.

Moraga I, Wernig G, Wilmes S, Gryshkova V, Richter C, Hong W . Tuning cytokine receptor signaling by re-orienting dimer geometry with surrogate ligands. Cell. 2015; 160(6):1196-208. PMC: 4766813. DOI: 10.1016/j.cell.2015.02.011. View

15.

Piehler J, Thomas C, Garcia K, Schreiber G . Structural and dynamic determinants of type I interferon receptor assembly and their functional interpretation. Immunol Rev. 2012; 250(1):317-34. PMC: 3986811. DOI: 10.1111/imr.12001. View

16.

Heink S, Yogev N, Garbers C, Herwerth M, Aly L, Gasperi C . Trans-presentation of IL-6 by dendritic cells is required for the priming of pathogenic T17 cells. Nat Immunol. 2016; 18(1):74-85. PMC: 5164931. DOI: 10.1038/ni.3632. View

17.

Gandhi H, Worch R, Kurgonaite K, Hintersteiner M, Schwille P, Bokel C . Dynamics and interaction of interleukin-4 receptor subunits in living cells. Biophys J. 2014; 107(11):2515-27. PMC: 4255262. DOI: 10.1016/j.bpj.2014.07.077. View

18.

Mendoza J, Schneider W, Hoffmann H, Vercauteren K, Jude K, Xiong A . The IFN-λ-IFN-λR1-IL-10Rβ Complex Reveals Structural Features Underlying Type III IFN Functional Plasticity. Immunity. 2017; 46(3):379-392. PMC: 5510750. DOI: 10.1016/j.immuni.2017.02.017. View

19.

Sarkar C, Lowenhaupt K, Horan T, Boone T, Tidor B, Lauffenburger D . Rational cytokine design for increased lifetime and enhanced potency using pH-activated "histidine switching". Nat Biotechnol. 2002; 20(9):908-13. DOI: 10.1038/nbt725. View

20.

Vogelsang J, Kasper R, Steinhauer C, Person B, Heilemann M, Sauer M . A reducing and oxidizing system minimizes photobleaching and blinking of fluorescent dyes. Angew Chem Int Ed Engl. 2008; 47(29):5465-9. DOI: 10.1002/anie.200801518. View