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PreprintCited by 0T-cell and B-cell ImmunologyProtein Tyrosine PhosphatasesProtein Kinase Regulation and GTPase Signaling

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Themis and Grb2 form a constitutive structural hub in T cell receptor signalling

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Savvides, Savvas, Clancy, Danielle, Sanjuan, Alba Sanz, Gillis, Elisabeth, Velghe, Imke, Droogenbroeck, Yana Van, Felix, Jan, Bloch, Yehudi, Cuadrado, Alvaro Furones, Merceron, Romain, Schenck, Stephan, Vandenabeele, Peter, Brunner, Janine, TAGHON, TOM, Elewaut, Dirk, Tougaard, Peter·2025·Nature Communications
DOI 10.21203/rs.3.rs-6042814/v1PMC PMC13381594

Preprint — published as 10.1038/s41467-026-73359-8 · Paper page

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Abstract

Positive selection of thymocytes is essential for laying the foundations of the mammalian immune system that include the T cell repertoire, self-tolerance, and prevention of autoimmunity. Themis, the archetypal member of a metazoan protein family featuring distinctive CABIT domains, crucially regulates thymocyte positive selection by linking signalling by the T cell receptor (TCR) to the linker of activation of TCR (LAT). Intriguingly, Themis has been proposed to function via a constitutive complex with the multifunctional adaptor Grb2. Although poised to represent a paradigm shift in our understanding of TCR signalling, the structural and mechanistic basis of such an assembly has remained enigmatic. Here, we present the cryo-EM structure of Themis in complex with Grb2, which reveals how the tandem CABIT domains of Themis engulf the C-terminal SH3 domain of Grb2 (Grb2 SH3C ) to enable its latching onto the proline rich sequence of Themis. The remaining two domains of Grb2 adopt at least three conformational poses set to interact with other binding partners such as Sos1. Structural insights from unbound Themis unmask the pronounced flexibility of the CABIT domains of Themis, which becomes ordered upon binding to Grb2 to create a binding hotspot for their constitutive complex. Indeed, Themis variants that abrogate interactions with Grb2 also fail to activate the tyrosine phosphatase SHP-1 after TCR stimulation, analogous to the functional phenotype of Themis-deficient cells. Collectively, our study draws the blueprint of the Themis-Grb2 complex as a dynamic structural hub in T cell development.

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Metadata

Authors
Savvides, S., Clancy, D., Sanjuan, A. S., Gillis, E., Velghe, I., Droogenbroeck, Y. V., Felix, J., Bloch, Y., Cuadrado, A. F., Merceron, R., Schenck, S., Vandenabeele, P., Brunner, J., TAGHON, T., Elewaut, D., & Tougaard, P.
Affiliations
  • Ghent University (BE)
  • VIB-UGent Center for Inflammation Research
  • Deutsches Elektronen-Synchrotron DESY (DE)
  • Vrije Universiteit Brussel (BE)
  • Ghent University Hospital (BE)
  • University of Copenhagen (DK)
Corresponding author
  • Savvides, Savvas
  • Clancy, Danielle
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2025-04-23
Volume
17
Issue
1
ISSN
2041-1723 (ISSN-L)
DOI
10.21203/rs.3.rs-6042814/v1
PMC
PMC13381594
OpenAlex
W7161848002
Cited by
0
References
1
FWCI
0.00
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by-nc-nd)
  • Ghent University Academic Bibliography (Ghent University) (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Fonds Wetenschappelijk Onderzoek (G0H1222N, 1219923N, G049820N)
  • Vlaams Instituut voor Biotechnologie (C0101)
  • Vlaamse regering
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Tyrosine Phosphatases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
THEMIS, Grb2, T cell receptor signaling, thymic selection, SH3 domain binding, protein-protein interaction, interdomain flexibility, adaptor protein complexes, T cell signaling
MeSH terms
Protein Domains, Animals, Humans, Protein Binding, Receptors, Antigen, T-Cell, T-Lymphocytes, Signal Transduction, src Homology Domains, Cryoelectron Microscopy, Intercellular Signaling Peptides and Proteins, Mice, GRB2 Adaptor Protein, Protein Tyrosine Phosphatase, Non-Receptor Type 6

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