SRC Homology 2 Domain-containing Leukocyte Phosphoprotein of 76 kDa (SLP-76) N-terminal Tyrosine Residues Regulate a Dynamic Signaling Equilibrium Involving Feedback of Proximal T-cell Receptor (TCR) Signaling

Engagement of the T cell receptor (TCR) of mature T lymphocytes can lead either to activation/proliferation responses or programmed cell death. To understand the molecular regulation of these two fundamentally different outcomes of TCR signaling, we investigated the participation of various components of the TCR-CD3 complex. We found that the TCR-zeta chain, while not absolutely required, was especially effective at promoting mature T cell apoptosis compared with the CD3 epsilon, gamma, or delta chains. We also carried out mutagenesis to address the role of the immunoreceptor tyrosine-based activation motifs (ITAMs) that are the principal signaling components found three times in the TCR-zeta chain and once in each of the CD3 epsilon, gamma, or delta chains. We found that the ability of the TCR-zeta chain to promote apoptosis results both from a quantitative effect of the presence of multiple ITAMs as well as qualitatively different contributions made by individual ITAMs. Apoptosis induced by single chain chimeras revealed that the first zeta ITAM stimulated greater apoptosis than the third zeta ITAM, and the second zeta ITAM was unable to trigger apoptosis. Because microheterogeneity in the amino acid sequence of the various ITAM motifs found in the TCR-zeta and CD3 chains predicts interactions with distinct src-homology-2-domain signaling proteins, our results suggest the possibility that individual ITAM motifs might play unique roles in TCR responses by engaging specific signaling pathways.

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A Novel Src Homology 2 Domain-containing Molecule, Src-like Adapter Protein-2 (SLAP-2), Which Negatively Regulates T Cell Receptor Signaling

Journal of Biological Chemistry ◽

10.1074/jbc.m110318200 ◽

2002 ◽

Vol 277 (21) ◽

pp. 19131-19138 ◽

Cited By ~ 31

Author(s):

Akhilesh Pandey ◽

Nieves Ibarrola ◽

Irina Kratchmarova ◽

Minerva M. Fernandez ◽

Stefan N. Constantinescu ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Adapter Protein ◽

Receptor Signaling ◽

Src Homology 2 ◽

T Cell Receptor Signaling ◽

Src Homology ◽

Src Homology 2 Domain ◽

Cell Receptor Signaling

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The ALX Src Homology 2 Domain Is Both Necessary and Sufficient to Inhibit T Cell receptor/CD28-mediated Up-regulation of RE/AP

Journal of Biological Chemistry ◽

10.1074/jbc.m404198200 ◽

2004 ◽

Vol 279 (39) ◽

pp. 40647-40652 ◽

Cited By ~ 12

Author(s):

Michael J. Shapiro ◽

Penda Powell ◽

Adanma Ndubuizu ◽

Chima Nzerem ◽

Virginia Smith Shapiro

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Src Homology 2 ◽

Src Homology ◽

Src Homology 2 Domain ◽

Necessary And Sufficient

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T cell receptor for antigen induces linker for activation of T cell–dependent activation of a negative signaling complex involving Dok-2, SHIP-1, and Grb-2

Journal of Experimental Medicine ◽

10.1084/jem.20060650 ◽

2006 ◽

Vol 203 (11) ◽

pp. 2509-2518 ◽

Cited By ~ 82

Author(s):

Shen Dong ◽

Béatrice Corre ◽

Eliane Foulon ◽

Evelyne Dufour ◽

André Veillette ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Critical Role ◽

Adaptor Proteins ◽

Cell Receptor ◽

Negative Control ◽

Inositol Polyphosphate ◽

Tcr Signaling ◽

Signaling Complex ◽

Src Homology 2 Domain

Adaptor proteins positively or negatively regulate the T cell receptor for antigen (TCR) signaling cascade. We report that after TCR stimulation, the inhibitory adaptor downstream of kinase (Dok)-2 and its homologue Dok-1 are involved in a multimolecular complex including the lipid phosphatase Src homology 2 domain–containing inositol polyphosphate 5′-phosphatase (SHIP)-1 and Grb-2 which interacts with the membrane signaling scaffold linker for activation of T cells (LAT). Knockdown of LAT and SHIP-1 expression indicated that SHIP-1 favored recruitment of Dok-2 to LAT. Knockdown of Dok-2 and Dok-1 revealed their negative control on Akt and, unexpectedly, on Zap-70 activation. Our findings support the view that Dok-1 and -2 are critical elements of a LAT-dependent negative feedback loop that attenuates early TCR signal. Dok-1 and -2 may therefore exert a critical role in shaping the immune response and as gatekeepers for T cell tolerance.

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Cytokine-Induced Src Homology 2 Protein (Cis) Promotes T Cell Receptor–Mediated Proliferation and Prolongs Survival of Activated T Cells

Journal of Experimental Medicine ◽

10.1084/jem.191.6.985 ◽

2000 ◽

Vol 191 (6) ◽

pp. 985-994 ◽

Cited By ~ 70

Author(s):

Suling Li ◽

Shangwu Chen ◽

Xiufeng Xu ◽

Anette Sundstedt ◽

Kajsa M. Paulsson ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Receptor ◽

T Cell Activation ◽

Cd4 T Cells ◽

Cell Activation ◽

Cell Receptor ◽

Cytokine Signaling ◽

Src Homology 2 ◽

Src Homology

Members of the suppressor of cytokine signaling (SOCS) family were discovered as negative regulators of cytokine signaling by inhibition of the Janus kinase–signal transducer and activator of transcription (Jak-STAT) pathway. Among them, cytokine-induced Src homology 2 (SH2) protein (CIS) was found to inhibit the interleukin 3– and erythropietin-mediated STAT5 signaling pathway. However, involvement of SOCS proteins in other signaling pathways is still unknown. This study shows that the expression of CIS is selectively induced in T cells after T cell receptor (TCR) stimulation. In transgenic mice, with selective expression of CIS in CD4 T cells, elevated CIS strongly promotes TCR-mediated proliferation and cytokine production in vitro, and superantigen-induced T cell activation in vivo. Forced expression of CIS also prolongs survival of CD4 T cells after TCR activation. Molecular events immediately downstream from the TCR are not changed in CIS-expressing CD4 T cells, but activation of mitogen-activated protein (MAP) kinase pathways by TCR stimulation is significantly enhanced. Together with the increased MAP kinase activation, a direct interaction of CIS and protein kinase Cθ was also demonstrated. These results suggest that CIS is one of the important regulators of TCR-mediated T cell activation. The functions of CIS, enhancing TCR signaling and inhibiting cytokine signaling, may be important in the regulation of immune response and homeostasis.

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Requirement of the Src Homology 2 Domain Protein Shb for T Cell Receptor-dependent Activation of the Interleukin-2 Gene Nuclear Factor for Activation of T Cells Element in Jurkat T Cells

Journal of Biological Chemistry ◽

10.1074/jbc.274.39.28050 ◽

1999 ◽

Vol 274 (39) ◽

pp. 28050-28057 ◽

Cited By ~ 29

Author(s):

Cecilia K. Lindholm ◽

Erik Gylfe ◽

Weiguo Zhang ◽

Lawrence E. Samelson ◽

Michael Welsh

Keyword(s):

T Cells ◽

T Cell Receptor ◽

Interleukin 2 ◽

Cell Receptor ◽

Nuclear Factor ◽

Jurkat T Cells ◽

Src Homology 2 ◽

Src Homology ◽

Domain Protein ◽

Src Homology 2 Domain

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Involvement of Lat, Gads, and Grb2 in Compartmentation of Slp-76 to the Plasma Membrane

Journal of Experimental Medicine ◽

10.1084/jem.192.6.847 ◽

2000 ◽

Vol 192 (6) ◽

pp. 847-856 ◽

Cited By ~ 57

Author(s):

Masamichi Ishiai ◽

Mari Kurosaki ◽

Kazunori Inabe ◽

Andrew C. Chan ◽

Kazuo Sugamura ◽

...

Keyword(s):

B Cell ◽

T Cell Receptor ◽

Adaptor Proteins ◽

Cell Receptor ◽

B Cell Receptor ◽

Cross Linking ◽

Src Homology 2 ◽

Src Homology ◽

The Difference ◽

Src Homology 2 Domain

B cell linker protein (BLNK) and Src homology 2 domain–containing leukocyte protein of 76 kD (SLP-76) are adaptor proteins required for B cell receptor (BCR) and T cell receptor function, respectively. Here, we show that expression of SLP-76 cannot reconstitute BCR function in Zap-70+BLNK− B cells. This could be attributable to inability of SLP-76 to be recruited into glycolipid-enriched microdomains (GEMs) after antigen receptor cross-linking. Supporting this idea, the BCR function was restored when a membrane-associated SLP-76 chimera was enforcedly localized to GEMs. Moreover, we demonstrate that addition of both linker for activation of T cells (LAT) and Grb2-related adaptor downstream of Shc (Gads) to SLP-76 allow SLP-76 to be recruited into GEMs, whereby the BCR function is reconstituted. The Gads function was able to be replaced by overexpression of Grb2. In contrast to SLP-76, BLNK did not require Grb2 families for its recruitment to GEMs. Hence, these data suggest a functional overlap between BLNK and SLP-76, while emphasizing the difference in requirement for additional adaptor molecules in their targeting to GEMs.

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Phosphorylation of multiple CD3 zeta tyrosine residues leads to formation of pp21 in vitro and in vivo. Structural changes upon T cell receptor stimulation.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)50741-4 ◽

1992 ◽

Vol 267 (5) ◽

pp. 3375-3381 ◽

Cited By ~ 2

Author(s):

S Koyasu ◽

D.J. McConkey ◽

L.K. Clayton ◽

S Abraham ◽

B Yandava ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Structural Changes ◽

Cell Receptor ◽

Receptor Stimulation ◽

Tyrosine Residues

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Usp12 stabilizes the T-cell receptor complex at the cell surface during signaling

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1521763113 ◽

2016 ◽

Vol 113 (6) ◽

pp. E705-E714 ◽

Cited By ~ 23

Author(s):

Akhee S. Jahan ◽

Maxime Lestra ◽

Lee Kim Swee ◽

Ying Fan ◽

Mart M. Lamers ◽

...

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Posttranslational Modifications ◽

Protein Function ◽

Adaptor Proteins ◽

Cell Receptor ◽

Surface Expression ◽

Jurkat Cells ◽

Tcr Signaling ◽

Primary Mouse

Posttranslational modifications are central to the spatial and temporal regulation of protein function. Among others, phosphorylation and ubiquitylation are known to regulate proximal T-cell receptor (TCR) signaling. Here we used a systematic and unbiased approach to uncover deubiquitylating enzymes (DUBs) that participate during TCR signaling in primary mouse T lymphocytes. Using a C-terminally modified vinyl methyl ester variant of ubiquitin (HA-Ub-VME), we captured DUBs that are differentially recruited to the cytosol on TCR activation. We identified ubiquitin-specific peptidase (Usp) 12 and Usp46, which had not been previously described in this pathway. Stimulation with anti-CD3 resulted in phosphorylation and time-dependent translocation of Usp12 from the nucleus to the cytosol. Usp12−/− Jurkat cells displayed defective NFκB, NFAT, and MAPK activities owing to attenuated surface expression of TCR, which were rescued on reconstitution of wild type Usp12. Proximity-based labeling with BirA-Usp12 revealed several TCR adaptor proteins acting as interactors in stimulated cells, of which LAT and Trat1 displayed reduced expression in Usp12−/− cells. We demonstrate that Usp12 deubiquitylates and prevents lysosomal degradation of LAT and Trat1 to maintain the proximal TCR complex for the duration of signaling. Our approach benefits from the use of activity-based probes in primary cells without any previous genome modification, and underscores the importance of ubiquitin-mediated regulation to refine signaling cascades.

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