Interactomics: complexes that assemble around different nodes of the TCR signaling cascade in primary CD4+ T cells

We analyze by mass spectrometry signalling complexes that form around canonical proteins used by the proximal TCR signal transduction pathway, and monitor their dynamic of assembly in the first minutes following engagement of the TCR.

In collaboration with B. Malissen and R. Roncagalli (CIML, Marseille), we use KI mouse model expressing different bait proteins of the TCR pathway with a C-terminal One-Strep-tag to enrich endogenous signaling complexes at different time points following TCR activation. Time resolved proteomics provides dynamics information (see for example the formation of the VAV1 interactome) and insights into the mechanisms of action of several proteins.

References

Reginald et al. Revisiting the Timing of Action of the PAG Adaptor Using Quantitative Proteomics Analysis of Primary T Cells. J. Immunol. 2015, 195 (11), 5472–5481. https://doi.org/10.4049/jimmunol.1501300.

 

Voisinne et al. Co-Recruitment Analysis of the CBL and CBLB Signalosomes in Primary T Cells Identifies CD5 as a Key Regulator of TCR-Induced Ubiquitylation. Mol. Syst. Biol. 2016, 12 (7), 876. https://doi.org/10.15252/msb.20166837.

 

Voisinne et al. CD5, an Undercover Regulator of TCR Signaling. Front. Immunol. 2018, 9. https://doi.org/10.3389/fimmu.2018.02900.

 

Gaud et al. The Costimulatory Molecule CD226 Signals through VAV1 to Amplify TCR Signals and Promote IL-17 Production by CD4+ T Cells. Sci. Signal. 2018, 11 (538). https://doi.org/10.1126/scisignal.aar3083.

 

Voisinne et al. Quantitative Interactomics in Primary T Cells Unveils TCR Signal Diversification Extent and Dynamics. Nat. Immunol. 2019, 20 (11), 1530–1541. https://doi.org/10.1038/s41590-019-0489-8.

 

Blaize et al. CD5 Signalosome Coordinates Antagonist TCR Signals to Control the Generation of Treg Cells Induced by Foreign Antigens. Proc. Natl. Acad. Sci. U. S. A. 2020, 117 (23), 12969–12979. https://doi.org/10.1073/pnas.1917182117.

 

Zhai et al. Opposing Regulatory Functions of the TIM3 (HAVCR2) Signalosome in Primary Effector T Cells as Revealed by Quantitative Interactomics. Cell. Mol. Immunol. 2020. https://doi.org/10.1038/s41423-020-00575-7.

 

Mori et al. The T Cell CD6 Receptor Operates a Multitask Signalosome with Opposite Functions in T Cell Activation. J. Exp. Med. 2021, 218 (2). https://doi.org/10.1084/jem.20201011.