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A global analysis of cross-talk in a mammalian cellular signalling network

Nature Cell Biology volume 8, pages 571–580 (2006)Cite this article

Abstract

Cellular information processing requires the coordinated activity of a large network of intracellular signalling pathways. Cross-talk between pathways provides for complex non-linear responses to combinations of stimuli, but little is known about the density of these interactions in any specific cell. Here, we have analysed a large-scale survey of pathway interactions carried out by the Alliance for Cellular Signalling (AfCS) in RAW 264.7 macrophages. Twenty-two receptor-specific ligands were studied, both alone and in all pairwise combinations, for Ca2+ mobilization, cAMP synthesis, phosphorylation of many signalling proteins and for cytokine production. A large number of non-additive interactions are evident that are consistent with known mechanisms of cross-talk between pathways, but many novel interactions are also revealed. A global analysis of cross-talk suggests that many external stimuli converge on a relatively small number of interaction mechanisms to provide for context-dependent signalling.

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Figure 1: The single ligand screen in RAW 264.7 cells.
Figure 2: Non-additivity between pairs of ligands applied to RAW 264.7 cells in all four experimental assays.
Figure 3: A subset of the double ligand screen in RAW 264.7 cells.
Figure 4: Testing the predicted interaction agent between calcium and cAMP.
Figure 5: The complete double ligand screen in RAW 264.7 cells describing non-additivity in the secretion of cytokines in response to all 231 pairwise combinations of 22 input ligands.
Figure 6: Testing the predicted interaction agent between Gαs–GPCRs and TLRs.
Figure 7: A schematic compilation of input–output relationships observed in the single and double-ligand screens.

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Acknowledgements

We are indebted to the many individuals from several academic institutes that are members of the AfCS scientific team for data collection, for database construction and for providing general help and advice. this work would not have been possible without the transparent and efficient public access to raw experimental data supported by the AfCS effort. We also thank members of the AfCS steering committee and the Ranganathan lab for discussion and critical reading of the manuscript. Work on this project was supported by contributions from public and private sources, including the NIGMS Glue Grant Initiative (U54 GM062114). A complete listing of the AfCS sponsors can be found at http://www.signalling-gateway.org/aboutus/sponsors.html. R.R. acknowledges support from the Mallinckrodt Scholar award, the Keck Future Initiatives award and the Robert A. Welch Foundation, and is an investigator of the Howard Hughes Medical Institute. P.C.S. acknowledges support from the Robert A. Welch Foundation and is the Alfred and Mabel Gilman Chair in molecular pharmacology.

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Authors and Affiliations

  1. Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, 75390–-9050, TX, USA

    Madhusudan Natarajan, Keng-Mean Lin, Robert C. Hsueh, Paul C. Sternweis & Rama Ranganathan

  2. The Alliance for Cellular Signalling, University of Texas Southwestern Medical Center, Dallas, 75390, TX, USA

    Madhusudan Natarajan, Keng-Mean Lin, Robert C. Hsueh & Paul C. Sternweis

  3. Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, 75390–9050, TX, USA

    Rama Ranganathan

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Correspondence to Rama Ranganathan.

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Supplementary Figures S1, S2, S3, S4, Supplementary Tables S1, S2, Supplementary Methods and Supplementary References (PDF 1397 kb)

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Natarajan, M., Lin, KM., Hsueh, R. et al. A global analysis of cross-talk in a mammalian cellular signalling network. Nat Cell Biol 8, 571–580 (2006). https://doi.org/10.1038/ncb1418

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