Abstract

Receptor-interacting protein kinase-1 (RIPK1), a master regulator of cell fate decisions, was identified as a direct substrate of MAPKAP kinase-2 (MK2) by phosphoproteomic screens using LPS-treated macrophages and stress-stimulated embryonic fibroblasts. p38MAPK/MK2 interact with RIPK1 in a cytoplasmic complex and MK2 phosphorylates mouse RIPK1 at Ser321/336 in response to pro-inflammatory stimuli, such as TNF and LPS, and infection with the pathogen Yersinia enterocolitica. MK2 phosphorylation inhibits RIPK1 autophosphorylation, curtails RIPK1 integration into cytoplasmic cytotoxic complexes, and suppresses RIPK1-dependent apoptosis and necroptosis. In Yersinia-infected macrophages, RIPK1 phosphorylation by MK2 protects against infection-induced apoptosis, a process targeted by Yersinia outer protein P (YopP). YopP suppresses p38MAPK/MK2 activation to increase Yersinia-driven apoptosis. Hence, MK2 phosphorylation of RIPK1 is a crucial checkpoint for cell fate in inflammation and infection that determines the outcome of bacteria–host cell interaction.

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Acknowledgements

We would like to acknowledge K. A. Fitzgerald, M. A. Kelliher, A. T. Ting and W. Brune for the gift of RIPK1-deficient cell lines, G. Tiegs and C. Wegscheid for providing the Tnfr1−/− mice, A. Pich for MS analysis, G. Evan, V. M. Dixit, J. Tschopp, and M. Treier for sharing expression vectors, A. Schambach for providing pLBID lentiviral vector, M. Windheim for discussion of results, K. Laaß for experimental support and T. Yakovleva for technical help. We thank A. Gossler for critical reading of the manuscript, J. C. Silva for help with phosphoproteomics, and I. Braren and the UKE HEXT Vector Facility for establishment of a lentiviral transduction system. This study was supported by grants from the Deutsche Forschungsgemeinschaft DFG to K.R. (Ru788/3-2 and Ru788/6-1) and M.G. (SFB566, TP B12).

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Author notes

    • Manoj B. Menon
    •  & Julia Gropengießer

    These authors contributed equally to this work.

    • Anne Deuretzbacher

    Present address: Department of Internal Medicine II, Division of Hematology, University Hospital of Würzburg, Würzburg 97080, Germany.

Affiliations

  1. Institute of Cell Biochemistry, Hannover Medical School, Hannover 30625, Germany

    • Manoj B. Menon
    • , Jessica Fischer
    • , Juri Lafera
    • , Natalia Ronkina
    • , Alexey Kotlyarov
    •  & Matthias Gaestel
  2. Institute for Medical Microbiology, Virology and Hygiene, University Medical Center Eppendorf, Hamburg 20246, Germany

    • Julia Gropengießer
    • , Lena Novikova
    • , Anne Deuretzbacher
    • , Hanna Schimmeck
    • , Nicole Czymmeck
    • , Martin Aepfelbacher
    •  & Klaus Ruckdeschel

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Contributions

M.B.M., J.G., A.K., M.G. and K.R. conceived the experiments and analysed the results. M.B.M., J.G., J.F., L.N., A.D., J.L., H.S., N.C. and N.R. performed the experiments. M.A. provided expertise and feedback. M.G. and K.R. secured funding. M.B.M., M.G. and K.R. wrote the manuscript.

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The authors declare no competing financial interests.

Corresponding authors

Correspondence to Matthias Gaestel or Klaus Ruckdeschel.

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https://doi.org/10.1038/ncb3614

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