Negative Regulation of T Cell Antigen Receptor-mediated Crk-L-C3G Signaling and Cell Adhesion by Cbl-b

Zhang, Wenying; Shao, Yinlin; Fang, Deyu; Huang, Jianyong; Jeon, Myung Shin; Liu, Yun‐Cai

doi:10.1074/jbc.m212671200

Cited by 69 publications

(63 citation statements)

References 29 publications

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“…Previous work has shown that CRK proteins signal through the RAP1 GEF C3G to regulate RAP1 activation (4,13,14,28). We now show that this pathway is important for chemokineinduced T cell adhesion and migration.…”

Section: Discussionsupporting

confidence: 65%

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CRK proteins selectively regulate T cell migration into inflamed tissues

Huang¹,

Clarke²,

Karimi³

et al. 2015

J. Clin. Invest.

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R e s e a R c h a R t i c l e1 0 2 0 jci.org Volume 125 Number 3 March 2015 therapeutic implications, since they can carry out graftversus-leukemia (GVL) responses with minimal GVHD. Results Generation and characterization of T cell-specific CRK T cells (data not shown).Western blotting of purified CD4 + T cells from Dko and WT mice revealed that levels of CRKI, CRKII, and CRKL in the mutant T cells were reduced by at least 95% ( Figure 1A and data not shown), and flow cytometric analysis confirmed loss of CRK protein expression (Figure 1, B and C).CRK/CRKL Dko mice were born at Mendelian ratios, and spleen, thymus, and lymph nodes exhibited normal cellularity (data not shown). Thymocyte populations were similar in WT and Dko mice, indicating that T cell development proceeded normally ( Figure 1D). Peripheral lymphoid organs showed no differences in proportions of CD4 + and CD8 + T cells, naive (CD62L hi CD44 lo ), memory (CD62L lo CD44 hi ), or activated (CD69 hi ) T cell subsets (Figure 1, D and E, and data not shown). Thus, these mice represented a suitable source of mature CRK/CRKL Dko T cells for functional studies. CRK/CRKL-deficient T cells show defects in adhesion and polarization.Since CRK proteins control adhesion in non-hematopoietic cells (4), we first asked whether integrin-dependent adhesion is defective in CRK/CRKL Dko T cells. On plates coated with ICAM-1, the ligand for the β 2 integrin LFA-1, WT preactivated CD4 + T cells showed about 10% basal binding, which was increased 2-to 5-fold after stimulation with the chemokines CXCL12 or CCL21, or anti-CD3 (Figure 2A). CRK/CRKL Dko T cells showed a significant reduction in adhesion to ICAM-1 under both basal and stimulated conditions. Indeed, chemokine stimulation induced almost no increased adhesion in these cells. Treatment with PMA bypassed the defect. This is most likely because the defect in CRK/CRKL Dko cells lies upstream of PKC signaling in the pathway leading to integrin activation, though PKC activation could also drive a parallel

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Section: Discussionsupporting

confidence: 65%

“…This was somewhat surprising, since CRK proteins have previously been implicated in TCR signaling (13,28), and since we observed decreased adhesion in response to anti-TCR stimulation in CRK/CRKL-deficient T cells ( Figure 1 0 3 0 jci.org Volume 125 Number 3 March 2015…”

Section: Methodsmentioning

confidence: 84%

CRK proteins selectively regulate T cell migration into inflamed tissues

Huang¹,

Clarke²,

Karimi³

et al. 2015

J. Clin. Invest.

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“…Cbl-b-dependent ubiquitylation of CrK-L prevents its association with C3G, therefore, inhibiting CrkL/C3G-driven clustering of cell adhesion molecules, in particular LFA-1 [39]. In accordance with early studies showing Cbl-b upregulation in anergized T cells, Cbl-b À/À T cells are resistant to ionomycin-induced anergy [34,40].…”

Section: Physiological and Molecular Roles Of E3 Ligases In T-cell Tosupporting

confidence: 73%

“…This results in the inhibition of Akt and PKCy-mediated NFkB activation [37], and additionally, prevents Vav1-mediated cytoskeleton rearrangements required for receptor clustering and synapse formation [38]. Furthermore, Cbl-b has been reported to destabilize the immune synapse by interfering with the CrkL/C3G signaling pathway [39].…”

Section: Physiological and Molecular Roles Of E3 Ligases In T-cell Tomentioning

confidence: 99%

E3 ubiquitin ligases in T‐cell tolerance

Paolino

Penninger

2009

Eur J Immunol

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The immune system uses several mechanisms of central and peripheral tolerance in order to prevent the activation of T lymphocytes toward self-antigens. Although the importance of immune self-tolerance has been established for a long time, some essential cellular and molecular mechanisms of T-cell tolerance have only been recently revealed. Once thought to be a recycling system, protein ubiquitylation by E3 ligases has now emerged as a regulated and crucial modulator of immune responses, and more importantly as a key signaling pathway involved in T-cell tolerance. In this review, we highlight our current understanding of the transcriptional and molecular signaling mechanisms involved in ubiquitylation-mediated T-cell tolerance.Key words: Autoimmunity . Cbl-b . E3 ligase . T-cell tolerance . Ubiquitylation IntroductionUpon activation, the immune system orchestrates robust and potentially destructive responses intended to eliminate the immunogenic antigen. Thus, immune system activation is under stringent control to prevent a detrimental immune response against self-antigens. The processes that ensure unresponsiveness toward self-antigens are known as immunological tolerance and, in their absence or failure, autoimmunity occurs [1]. Central tolerance mechanisms operate in the thymus to eliminate potentially self-reactive thymocytes and generate (T reg ) suppressor cells [2]. In addition, several peripheral tolerance mechanisms act to prevent self-reactivity once mature T cells have reached the periphery. These peripheral mechanisms of tolerance include the following: (i) immunological ignorance to selfantigens expressed at low levels or in immunoprivileged sites; (ii) active immunosuppression by Foxp31 T reg cells; (iii) activation-induced cell death of autoreactive T cells and (iv) induction of a state of unresponsiveness known as T-cell anergy [3].During the past few years, extensive research has revealed new insights into the cellular and molecular mechanisms necessary to induce and maintain T-cell tolerance. In particular, ubiquitylation of proteins by E3 ligases has emerged as a novel and indispensable signaling pathway that regulates T-cell tolerance toward self-antigens [4][5][6]. The ubiquitylation processUbiquitylation is an important mechanism of post-translational modification of proteins by which the 76-aa polypeptide ubiquitin, Ub, is covalently attached to a substrate protein. This process occurs in a stepwise manner, requiring the participation of three enzymes: a ubiquitin-activating enzyme E1, responsible for the first binding and activation of the Ub; a second ubiquitinconjugating enzyme E2, which receives the activated Ub from the E1; and a third ubiquitin-ligase E3 that catalyzes the final covalent transfer of the Ub from the E2 enzyme to the protein substrate [7]. Once thought to only mediate proteosomal degradation, the ubiquitylation of a protein can have multiple effects, including altered subcellular localization, regulation of protein-protein interactions, and functional modulation. The fa...

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“…Conversely, we found that in BMMC, the absence of Cbl-b did not inhibit Ag-induced Fc⑀RI internalization, nor did it have an effect on receptor degradation after 6-h Ag stimulation (data not shown). Furthermore, in WT mast cells, 6-h Ag incubation resulted in even a slight increase in Cbl-b protein expression (data not shown) similar to that observed after TCR stimulation of primary T cells (42). Fc⑀RI stimulation induces the ubiquitination of Syk (43).…”

Section: Discussionmentioning

confidence: 54%

Inactivation of c-Cbl or Cbl-b Differentially Affects Signaling from the High Affinity IgE Receptor

Zhang

Chiang

Hodes

et al. 2004

The Journal of Immunology

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The Cbl family of proteins negatively regulate signaling from tyrosine kinase-coupled receptors. Among the three members of this family, only c-Cbl and Cbl-b are expressed in hemopoietic cells. To examine the role of c-Cbl and Cbl-b in FcεRI signaling, mast cell cultures from wild-type, c-Cbl−/−, and Cbl-b−/− mice were generated. Cell growth rates and cell surface expression of FcεRI were similar in the different cell populations. Compared with control cells, Cbl-b inactivation resulted in increases in FcεRI-induced Ca2+ response and histamine release. FcεRI-induced tyrosine phosphorylation of total cellular proteins, Syk, and phospholipase C-γ was also enhanced by Cbl-b deficiency, whereas receptor-initiated phosphorylation of Vav, JNK, and p38 kinases was not changed in these cells. In contrast to Cbl-b, c-Cbl deficiency had no detectable effect on FcεRI-induced histamine release or on the phosphorylation of total cellular proteins or Syk. The absence of c-Cbl increased the phosphorylation of ERK after receptor stimulation, but resulted in slightly reduced p38 phosphorylation and Ca2+ response. These results suggest that Cbl-b and c-Cbl have divergent effects on FcεRI signal transduction and that Cbl-b, but not c-Cbl, functions as a negative regulator of FcεRI-induced degranulation.

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Negative Regulation of T Cell Antigen Receptor-mediated Crk-L-C3G Signaling and Cell Adhesion by Cbl-b

Cited by 69 publications

References 29 publications

CRK proteins selectively regulate T cell migration into inflamed tissues

CRK proteins selectively regulate T cell migration into inflamed tissues

E3 ubiquitin ligases in T‐cell tolerance

Inactivation of c-Cbl or Cbl-b Differentially Affects Signaling from the High Affinity IgE Receptor

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