Journal article

A regulatory region on RIPK2 is required for XIAP binding and NOD signaling activity

Valentin J Heim, Laura F Dagley, Che A Stafford, Fynn M Hansen, Elise Clayer, Aleksandra Bankovacki, Andrew Webb, Isabelle S Lucet, John Silke, Ueli Nachbur

EMBO REPORTS | WILEY | Published : 2020

Abstract

Signaling via the intracellular pathogen receptors nucleotide-binding oligomerization domain-containing proteins NOD1 and NOD2 requires receptor interacting kinase 2 (RIPK2), an adaptor kinase that can be targeted for the treatment of various inflammatory diseases. However, the molecular mechanisms of how RIPK2 contributes to NOD signaling are not completely understood. We generated FLAG-tagged RIPK2 knock-in mice using CRISPR/Cas9 technology to study NOD signaling mechanisms at the endogenous level. Using cells from these mice, we were able to generate a detailed map of post-translational modifications on RIPK2. Similar to other reports, we did not detect ubiquitination of RIPK2 lysine 209 ..

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Grants

Awarded by NHMRC


Awarded by ARC fellowship


Awarded by Australian Government NHMRC IRIISS


Awarded by European Union's Framework Programme for Research and Innovation Horizon 2020 (2014-2020) under the Marie Sklodowska-Curie Grant


Awarded by LMU Munich's Institutional Strategy LMUexcellent


Funding Acknowledgements

We thank Prof. C. Day for the GST-BIR2 constructs and C. Gatt, K.McKenzie, T. Ballinger, and C. Epifanio for assistance with animal work and the MAGEC facility of WEHI for the generation of affinity-tagged RIPK2 mice. The work was supported by NHMRC grants 1046986, 1057888 and fellowships, 541901, 1058190 to JS and an ARC fellowship, FT130100166, to UN. This work was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS (#9000220). This project has received funding from the European Union's Framework Programme for Research and Innovation Horizon 2020 (2014-2020) under the Marie Sklodowska-Curie Grant Agreement No. 754388 and from LMU Munich's Institutional Strategy LMUexcellent within the framework of the German Excellence Initiative (No. ZUK22).