Gasp, a Grb2-associating protein, is critical for positive selection of thymocytes

Patrick, Michael S.; Oda, Hiroyo; Hayakawa, Kunihiro; Satoh, Yoshinori; Eshima, Koji; Kirikae, Teruo; Iemura, Shun Ichiro; Shirai, Mutsunori; Abe, Takaya; Natsume, Tohru; Sasazuki, Takehiko; Suzuki, Harumi

doi:10.1073/pnas.0908593106

Cited by 64 publications

(117 citation statements)

References 41 publications

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“…Despite the absence of strong biochemical evidence, it was presumed that Themis1 likely functions as a positive regulator of TCR signaling because several defects in Themis1 −/− mice, such as a marked impairment of both positive and negative selection, were consistent with a deficiency in TCR signaling (2)(3)(4)(5)(6). It was subsequently shown that Themis1 interacts with Vav1 and that Vav1 phosphorylation is reduced in Themis1 −/− thymocytes, results that also implied a positive role for Themis1 in TCR signaling (9).…”

Section: Discussionmentioning

confidence: 99%

“…We and others identified the previously uncharacterized protein Themis1 as being required for the positive selection and maturation of T cells in the thymus (2)(3)(4)(5)(6). Themis1 lacks any discernable catalytic domain, but it contains two previously uncharacterized cysteine-based globular domains (named "CABIT") of unknown structure and function (3).…”

Section: Introductionmentioning

confidence: 99%

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Themis1 enhances T cell receptor signaling during thymocyte development by promoting Vav1 activity and Grb2 stability

et al. 2016

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The T cell signaling protein Themis1 is essential for the positive and negative selection of thymocytes in the thymus. Although the developmental defect that results from the loss of Themis1 suggests that it enhances T cell receptor (TCR) signaling, Themis1 also recruits Src homology 2 domain-containing phosphatase-1 (SHP-1) to the vicinity of TCR signaling complexes, suggesting that it has an inhibitory role in TCR signaling. We used TCR signaling reporter mice and quantitative proteomics to explore the role of Themis1 in developing T cells. We found that Themis1 acted mostly as a positive regulator of TCR signaling in vivo when receptors were activated by positively selecting ligands. Proteomic analysis of the Themis1 interactome identified SHP-1, the TCR-associated adaptor protein Grb2, and the guanine nucleotide exchange factor Vav1 as the principal interacting partners of Themis1 in isolated mouse thymocytes. Analysis of TCR signaling in Themis1-deficient and Themis1-overexpressing mouse thymocytes demonstrated that Themis1 promoted Vav1 activity both in vitro and in vivo. The reduced activity of Vav1 and the impaired T cell development in Themis1 −/− mice were due in part to increased degradation of Grb2, which suggests that Themis1 is required to maintain the steady-state abundance of Grb2 in thymocytes. Together, these data suggest that Themis1 acts as a positive regulator of TCR signaling in developing T cells, and identify a mechanism by which Themis1 regulates thymic selection.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Themis1 enhances T cell receptor signaling during thymocyte development by promoting Vav1 activity and Grb2 stability

et al. 2016

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“…THEMIS expression is high in double-positive (DP) thymocytes, with a marked downregulation after positive selection and in peripheral T cells. Whereas knockout mice did not show hallmarks of autoimmunity (3,5), the recently described BN m rat strain has linked a frameshift mutation in the Themis gene to inflammatory bowel disease caused by defective regulatory T cell function (6). THEMIS is a highly conserved, 73-kDa cytoplasmic protein without any obvious catalytic or protein-protein interaction domains.…”

Section: T Hymocyte-expressed Molecule Involved In Selection (Themis)mentioning

confidence: 99%

“…Bioinformatics analysis predicts a tandem repeat of a novel, cysteine-containing globular domain (cysteine-containing all b in THEMIS [CABIT]) found in a number of metazoan proteins (2). A putative bipartite nuclear localization sequence is present within the CABIT-2 domain of THEMIS, but no nuclear translocation was detected upon TCR ligation (3,5). The C-terminal end of THEMIS, predicted to contain little or no secondary structure, harbors two proline-rich regions (PRRs).…”

Section: T Hymocyte-expressed Molecule Involved In Selection (Themis)mentioning

confidence: 99%

GRB2-Mediated Recruitment of THEMIS to LAT Is Essential for Thymocyte Development

Paster

Brockmeyer

et al. 2013

The Journal of Immunology

100

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Thymocyte-expressed molecule involved in selection (THEMIS) is a recently identified regulator of thymocyte positive selection. THEMIS’s mechanism of action is unknown, and whether it has a role in TCR-proximal signaling is controversial. In this article, we show that THEMIS and the adapter molecule growth factor receptor–bound protein 2 (GRB2) associate constitutively through binding of a conserved PxRPxK motif within the proline-rich region 1 of THEMIS to the C-terminal SH3-domain of GRB2. This association is indispensable for THEMIS recruitment to the immunological synapse via the transmembrane adapter linker for activation of T cells (LAT) and for THEMIS phosphorylation by Lck and ZAP-70. Two major sites of tyrosine phosphorylation were mapped to a YY-motif close to proline-rich region 1. The YY-motif was crucial for GRB2 binding, suggesting that this region of THEMIS might control local phosphorylation-dependent conformational changes important for THEMIS function. Finally, THEMIS binding to GRB2 was required for thymocyte development. Our data firmly assign THEMIS to the TCR-proximal signaling cascade as a participant in the LAT signalosome and suggest that the THEMIS–GRB2 complex might be involved in shaping the nature of Ras signaling, thereby governing thymic selection.

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“…Alternatively, but not mutually exclusively, the role of Grb2 in T cell selection might be explained by its ability to recruit a THEMISGrb2-SHP1/2 complex to LAT in the immunological synapse, where THEMIS and SHP1/2 set the threshold for positive selection (6,7). In the absence of THEMIS or if the THEMIS binding site for Grb2 is mutated, positive selection is drastically impaired (5,7,33,34). The importance of the C-terminal SH3 domain of Grb2 that recruits THEMIS (6) is also highlighted by studies in which mutated forms of Grb2 were transduced into bone marrow cells from Grb2 fl/fl Lckcre tg mice via retroviral transduction.…”

Section: Discussionmentioning

confidence: 99%

Grb2 Is Important for T Cell Development, Th Cell Differentiation, and Induction of Experimental Autoimmune Encephalomyelitis

Radtke

Lacher

Szumilas

et al. 2016

The Journal of Immunology

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The small adaptor protein growth factor receptor–bound protein 2 (Grb2) modulates and integrates signals from receptors on cellular surfaces in inner signaling pathways. In murine T cells, Grb2 is crucial for amplification of TCR signaling. T cell–specific Grb2fl/fl Lckcretg Grb2-deficient mice show reduced T cell numbers due to impaired negative and positive selection. In this study, we found that T cell numbers in Grb2fl/fl CD4cretg mice were normal in the thymus and were only slightly affected in the periphery. Ex vivo analysis of CD4+ Th cell populations revealed an increased amount of Th1 cells within the CD4+ population of Grb2fl/fl CD4cretg mice. Additionally, Grb2-deficient T cells showed a greater potential to differentiate into Th17 cells in vitro. To test whether these changes in Th cell differentiation potential rendered Grb2fl/fl CD4cretg mice more prone to inflammatory diseases, we used the murine Th1 cell– and Th17 cell–driven model of experimental autoimmune encephalomyelitis (EAE). In contrast to our expectations, Grb2fl/fl CD4cretg mice developed a milder form of EAE. The impaired EAE disease can be explained by the reduced proliferation rate of Grb2-deficient CD4+ T cells upon stimulation with IL-2 or upon activation by allogeneic dendritic cells, because the activation of T cells by dendritic cells and the subsequent T cell proliferation are known to be crucial factors for the induction of EAE. In summary, Grb2-deficient T cells show defects in T cell development, increased Th1 and Th17 cell differentiation capacities, and impaired proliferation after activation by dendritic cells, which likely reduce the clinical symptoms of EAE.

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Gasp, a Grb2-associating protein, is critical for positive selection of thymocytes

Cited by 64 publications

References 41 publications

Themis1 enhances T cell receptor signaling during thymocyte development by promoting Vav1 activity and Grb2 stability

Themis1 enhances T cell receptor signaling during thymocyte development by promoting Vav1 activity and Grb2 stability

GRB2-Mediated Recruitment of THEMIS to LAT Is Essential for Thymocyte Development

Grb2 Is Important for T Cell Development, Th Cell Differentiation, and Induction of Experimental Autoimmune Encephalomyelitis

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