T Cell Development and T Cell Responses in Mice with Mutations Affecting Tyrosines 292 or 315 of the Zap-70 Protein Tyrosine Kinase

Magnan, A.; Bartolo, Vincenzo Di; Mura, Anne-Marie; Boyer, Catherine; Richelme, Mireille; Lin, Yongshun; Roure, Agnès; Gillet, Anne; Arrieumerlou, Cécile; Acuto, Oreste; Malissen, Bernard; Malissen, Marie

doi:10.1084/jem.194.4.491

Cited by 51 publications

(78 citation statements)

References 71 publications

(91 reference statements)

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“…Both ZAP70 Y292 and ZAP70 Y315/Syk Y352 phosphorylation patterns showed similar trends in the genotypic analysis, which was of interest because Y292 has been reported to play a role in attenuating the TCR signal and Y315 in the enhancement of ZAP70 function (26)(27)(28). The downstream signaling consequences of these two opposing regulatory effects of Y292 and Y315 is unclear, made more difficult because the phosphorylation signal obtained at Y315 also is capturing that of Syk Y352 because this anti-ZAP70 Ab cross-reacts with the Syk Y352 residue.…”

Section: Discussionmentioning

confidence: 84%

Allelic variant inCTLA4alters T cell phosphorylation patterns

Maier

Anderson

Jager

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Little is known regarding the functional effects of common autoimmune susceptibility variants on human immune cells. The SNP CT60 (rs3087243; A/G) located in the 3 UTR of the CTLA4 gene has been associated with autoimmune diseases. We examined a cohort of healthy individuals stratified by genotypes at CTLA4 to gain insight into the functional effects of allelic variation on T cell signaling. Using phospho-site-specific mAbs, we tested the hypothesis that the CT60 genotype at CTLA4 is associated with altered T cell antigen receptor (TCR) signaling in naive and/or memory T cells. By normalizing for the extent of the initial TCR signaling event at CD3 , we observed that the relative responsiveness to TCR stimulation as assessed by phosphorylation levels of downstream signaling molecules was altered in naive (CD4 ؉ CD45RA high ) and memory (CD4 ؉ CD45RA low ) T cells obtained from individuals with the disease-susceptibility allele at CTLA4. Thus, allelic variation associated with autoimmune disease can alter the signaling threshold of CD4 ؉ T cells. These experiments provide a rational approach for the dissection of T cell-susceptibility genes in autoimmune diseases.genotype ͉ human ͉ T cell antigen receptor signaling ͉ CTLA4 ͉ autoimmunity T he recent successful completion of several genome-wide association scans, together with the rapid advancement in highthroughput genotyping technologies, have resulted in the discovery of an ever-increasing number of genetic variants associated with susceptibility to human autoimmune diseases (1-5). However, because of the small genetic effects of these variants, compared with the strong genetic effects seen in Mendelian diseases, the study of genotypic variation in relation to phenotypes has been a substantial challenge. One well validated region associated with susceptibility to autoimmune disease harbors the CTLA4 gene on chromosome 2q33. An associated allele in this susceptibility region is the G allele of the SNP CT60 (rs3087243; A/G) in the CTLA4 gene region, which has been associated with risk to type 1 diabetes, Graves disease, autoimmune hypothyroidism, systemic lupus erythematosus, and Addison's disease (6-10).The expression of mRNA isoforms of CTLA4 has been investigated in relation to CTLA4 genotype in healthy controls. Moreover, the genotype-dependent difference in the expression of soluble CTLA4 transcripts appears to be T cell-specific (6, 10). In the nonobese diabetic (NOD) mouse, the orthologous CTLA4 region also is associated with autoimmune diabetes, and, similar to humans, an allelic variant causes the increased expression of a major splice isoform. In the NOD model, the isoform is ligandindependent CTLA-4 (11) and results in strongly inhibited T cell responses, as well as increased susceptibility to disease (12). Together these investigations in both the experimental model and in humans suggest that genetic variation in the CTLA4 gene region may have an important effect on T cell function because of altered T cell signaling.Although CTLA4 is important in aut...

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Section: Discussionmentioning

confidence: 84%

Allelic variant inCTLA4alters T cell phosphorylation patterns

Maier

Anderson

Jager

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…Studies on Zap70(Y315F) in cells and mice demonstrated that Tyr 315 is important for optimal activity of Zap70 during the development of T cells in the thymus and activation of mature T cells in the lymph nodes and peripheral blood (33,54,56). Expression of Zap70(Y315F) in Syk-deficient DT40 cells impaired the function of Zap70 in BCR signaling by reducing the BCR-induced tyrosine phosphorylation of Zap70, Vav, SH2 domain-containing leukocyte protein of 76 kDa, and Shc, and attenuating the BCRinduced NF-AT-regulated responses (33).…”

Section: Discussionmentioning

confidence: 99%

“…Some of the phosphotyrosine residues were implicated in the regulation of Zap70 catalytic activity (36, 45-47), while others, predominantly those located at the interdomain B region (see Fig. 2), were found to mediate interaction with distinct SH2-containing effector molecules (33,37,(47)(48)(49)(50)(51)(52)(53)(54)(55)(56). To test whether interdomain B-residing phosphotyrosyl residues are required for the interaction with CrkII, we used Cos-7 cells that transiently overexpress Myc-tagged WT Zap70 or Myc-tagged Zap70 ⌬265-331 proteins.…”

Section: Truncation Of the Interdomain B Region Abrogates The Abilitymentioning

confidence: 99%

T Cell Activation-Induced CrkII Binding to the Zap70 Protein Tyrosine Kinase Is Mediated by Lck-Dependent Phosphorylation of Zap70 Tyrosine 315

Gelkop

Gish

Babichev

et al. 2005

The Journal of Immunology

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The Zap70 protein tyrosine kinase controls TCR-linked signal transduction pathways and is critical for T cell development and responsiveness. Following engagement of TCR, the Zap70 undergoes phosphorylation on multiple tyrosine residues that are implicated in the regulation of its catalytic activity and interaction with signaling effector molecules downstream of the TCR. We have shown previously that the CT10 regulator of kinase II (CrkII) adapter protein interacts with tyrosine-phosphorylated Zap70 in TCR-engaged T cells, and now extend these studies to show that Tyr315 in the Zap70 interdomain B region is the site of interaction with CrkII. A point mutation of Tyr315 (Y315F) eliminated the CrkII-Zap70 interaction capacity. Phosphorylation of Tyr315 and Zap70 association with CrkII were both dependent upon the Lck protein tyrosine kinase. Previous studies demonstrated the Tyr315 is the Vav-Src homology 2 (SH2) binding site, and that replacement of Tyr315 by Phe impaired the function of Zap70 in TCR signaling. However, fluorescence polarization-based binding studies revealed that the CrkII-SH2 and the Vav-SH2 bind a phosphorylated Tyr315-Zap70-derived peptide with affinities of a similar order of magnitude (Kd of 2.5 and 1.02 μM, respectively). The results suggest therefore that the biological functions attributed to the association of Zap70 with Vav following T cell activation may equally reflect the association of Zap70 with CrkII, and further support a regulatory role for CrkII in the TCR-linked signal transduction pathway.

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“…Two of these tyrosine residues, Tyr-492 and Tyr-493 in human ZAP-70, are positioned in the activation loop of the catalytic domain, and their phosphorylation either by Lck (13,15) or by ZAP-70 itself in trans (17) contributes to the activation of ZAP-70 by stabilizing the open and extended conformation of the activation loop. Two regulatory tyrosine residues of the SH2-kinase linker, Tyr-315 and Try319, are known to become phosphorylated (16,17,19) when ZAP-70 is recruited to the TCR and have been implicated in an autoinhibitory mechanism controlling the catalytic activity of 16,17,[19][20][21][22].…”

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confidence: 99%

Stability of an autoinhibitory interface in the structure of the tyrosine kinase ZAP-70 impacts T cell receptor response

Deindl

Kadlecek

Cao

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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The delivery of signals from the activated T cell antigen receptor (TCR) inside the cell relies on the protein tyrosine kinase ZAP-70 (-associated protein of 70 kDa). A recent crystal structure of inactive full-length ZAP-70 suggests that a central interface formed by the docking of the two SH2 domains of ZAP-70 onto the kinase domain is crucial for suppressing catalytic activity. Here we validate the significance of this autoinhibitory interface for the regulation of ZAP-70 catalytic activity and the T cell response. For this purpose, we perform in vitro catalytic activity assays and binding experiments using ZAP-70 proteins purified from insect cells to examine activation of ZAP-70. Furthermore, we use cell lines stably expressing wild-type or mutant ZAP-70 to monitor proximal events in T cell signaling, including TCR-induced phosphorylation of ZAP-70 substrates, activation of the MAP kinase pathway, and intracellular Ca 2؉ levels. Taken together, our results directly correlate the stability of the autoinhibitory interface with the activation of these key events in the T cell response.activation ͉ catalytic activity ͉ conformation ͉ ITAM T he protein tyrosine kinase ZAP-70 (-associated protein of 70 kDa) (1) participates in the initial events of T cell antigen receptor (TCR) signaling and thus is of critical importance for the adaptive immune response (2-5). On activation of the TCR, 2 tyrosines in the immunoreceptor tyrosine-based activation motifs (ITAMs) of the CD3 or cytoplasmic tails of the TCR are phosphorylated by a Src family kinase member. Through the binding of 2 SH2 (Src homology 2) domains at the N terminus of ZAP-70 to these ITAM phosphotyrosines, ZAP-70 is then specifically recruited to a stimulated TCR. These 2 SH2 domains of ZAP-70 are tightly associated as a tandem unit with a precise structural organization that enables high-affinity ITAM peptide binding (6). In the very closely related Syk (spleen tyrosine) kinase, the destabilization of this structural organization by mutation results in both uncoupling of the 2 SH2 units and a dramatic reduction in ITAM binding affinity (7). Recruitment of ZAP-70 to the TCR results in activation of the catalytic domain located at the C terminus of ZAP-70 and signal propagation by phosphorylation of its downstream target molecules, including the ZAP-70 substrates LAT (linker for the activation of T cells) and SLP-76 (SH2 domain-containing leukocyte phosphoprotein of 76 kDa). Both LAT and SLP-76 are crucial adaptor proteins that, once phosphorylated, provide a platform for the assembly of signaling molecules (8, 9). Ultimately, these signaling events produce effector secretory responses, modulate transcription, and trigger T cell proliferation and differentiation.ZAP-70 plays critical roles in pre-TCR and TCR signaling, thymocyte development, and peripheral T cell function. The loss of ZAP-70 catalytic activity results in profound immunodeficiency, and reduced activity of ZAP-70 may result in autoimmunity (4, 10). Furthermore, the expression of ZAP-70 in B c...

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T Cell Development and T Cell Responses in Mice with Mutations Affecting Tyrosines 292 or 315 of the Zap-70 Protein Tyrosine Kinase

Cited by 51 publications

References 71 publications

Allelic variant inCTLA4alters T cell phosphorylation patterns

Allelic variant inCTLA4alters T cell phosphorylation patterns

T Cell Activation-Induced CrkII Binding to the Zap70 Protein Tyrosine Kinase Is Mediated by Lck-Dependent Phosphorylation of Zap70 Tyrosine 315

Stability of an autoinhibitory interface in the structure of the tyrosine kinase ZAP-70 impacts T cell receptor response

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