ZAP-70 in Signaling, Biology, and Disease

Au-Yeung, Byron B; Shah, Neel H.; Shen, Lin; Weiss, Arthur

doi:10.1146/annurev-immunol-042617-053335

Cited by 117 publications

(110 citation statements)

References 184 publications

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“…Using a model of ZAP-70-deficient mice, exhibiting an arrest in thymocyte differentiation at the DP stage, 58 we assessed whether intrathymic administration of an AAV8 vector could be used to efficiently achieve gene transfer in defective thymocytes. Importantly, 3 days after intrathymic transfer of the AAV2/AAV8-GFP vector used above (Fig 1, A), transduction of both DN and DP thymocytes was detected (Fig 1, D).…”

Section: Resultsmentioning

confidence: 99%

“…Intrathymic administration of all 3 serotypes led to transgene expression in all thymocyte subsets, but AAV8 exhibited the greatest gene transfer efficiency, resulting in transduction of up to 5% of all thymocytes. Using mice that are immunodeficient because of mutations in the zeta-associated protein of 70 kDa (ZAP-70) protein tyrosine kinase 58 as a paradigm, we found that intrathymic injection of an AAV2/ AAV8-ZAP-70 vector resulted in the development of gene-corrected mature thymocytes within 10 days of gene transfer. Concurrently, AAV8-ZAP-70 gene transfer promoted development of the thymic medulla, containing autoimmune regulator (AIRE)-expressing medullary thymic epithelial cells (mTECs), which mediate T-cell tolerance.…”

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confidence: 99%

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Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells

Pouzolles

Machado

Guilbaud

et al. 2020

Journal of Allergy and Clinical Immunology

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Background: Patients with T-cell immunodeficiencies are generally treated with allogeneic hematopoietic stem cell transplantation, but alternatives are needed for patients without matched donors. An innovative intrathymic gene therapy approach that directly targets the thymus might improve outcomes. Objective: We sought to determine the efficacy of intrathymic adeno-associated virus (AAV) serotypes to transduce thymocyte subsets and correct the T-cell immunodeficiency in a zetaassociated protein of 70 kDa (ZAP-70)-deficient murine model. Methods: AAV serotypes were injected intrathymically into wild-type mice, and gene transfer efficiency was monitored. ZAP-70 2/2 mice were intrathymically injected with an AAV8 vector harboring the ZAP70 gene. Thymus structure, immunophenotyping, T-cell receptor clonotypes, T-cell function, immune responses to transgenes and autoantibodies, vector copy number, and integration were evaluated. Results: AAV8, AAV9, and AAV10 serotypes all transduced thymocyte subsets after in situ gene transfer, with transduction of up to 5% of cells. Intrathymic injection of an AAV8-ZAP-70 vector into ZAP-70 2/2 mice resulted in a rapid thymocyte differentiation associated with the development of a thymic medulla. Strikingly, medullary thymic epithelial cells expressing the autoimmune regulator were detected within 10 days of gene transfer, correlating with the presence of functional effector and regulatory T-cell subsets with diverse T-cell receptor clonotypes in the periphery. Although thymocyte reconstitution was transient, gene-corrected peripheral T cells harboring approximately 1 AAV genome per cell persisted for more than 40 weeks, and AAV vector integration was detected. Conclusions: Intrathymic AAV-transduced progenitors promote a rapid restoration of the thymic architecture, with a single wave of thymopoiesis generating long-term peripheral T-cell function.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells

Pouzolles

Machado

Guilbaud

et al. 2020

Journal of Allergy and Clinical Immunology

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“…Binding to ITAMs triggers conformational changes that make ZAP-70 more accessible to phosphorylation by Lck and subsequently leads to ZAP-70 autophosphorylation and full activation (3)(4)(5). Activated ZAP-70 is released from ITAMs and proceeds to target its effectors, primarily membrane-bound scaffolding proteins LAT and SLP-76 (6).…”

Section: Introductionmentioning

confidence: 99%

T cell receptor-dependent S-acylation of ZAP-70 controls activation of T cells

Tewari

Bieerkehazhi

Fan

et al. 2020

Preprint

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AbstractZAP-70 is a cytoplasmic tyrosine kinase essential for T cell-mediated immune responses. Upon engagement of the T cell receptor, ZAP-70 is quickly recruited to the specialized plasma membrane domains, becomes activated and released to phosphorylate its laterally segregated downstream targets. A shift in ZAP-70 distribution at the plasma membrane is recognized as a critical step in T cell receptor signal transduction and amplification. However, the molecular mechanism supporting stimulation-dependent plasma membrane compartmentalization of ZAP-70 remains poorly understood. In this study, we identified previously uncharacterized reversible lipidation (S-acylation) of ZAP-70. We found that this post-translational modification of ZAP-70 is dispensable for its enzymatic activity. However, the lipidation-deficient mutant of ZAP-70 failed to propagate the T cell receptor signaling cascade suggesting that S-acylation is essential for ZAP-70 interaction with its protein substrates. The kinetics of ZAP-70 S-acylation were consistent with early T cell signaling events indicating that agonist-induced S-acylation is a part of the signaling mechanism controlling T cell activation and function.Significance StatementActivation of T cells is a critical part of the adaptive immune response to pathogen exposure. We found that ZAP-70, a regulatory protein essential for T cell activation, can undergo a post-translational modification with long chain fatty acids, known as S-acylation. In this report, we show that S-acylation of ZAP-70 is T cell receptor-dependent and required for its signaling function. We found that loss of ZAP-70 S-acylation resulted in T cell unresponsiveness to T cell receptor stimulation indicating that abnormalities in protein S-acylation can potentially contribute to the T cell immunodeficiency disorders.

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“…Although Syk and ZAP-70 share high sequence similarity, similar domain architecture and activated by the conceptually same mechanism(11, 19–21, 37, 39, 47, 48), yet these two kinases recruit to the membrane by a fundamentally different mechanism. In contrast to Syk, the tSH2 domain of ZAP-70 undergoes a biphasic structural transition while binding to the doubly-phosphorylated ITAM peptide.…”

Section: Resultsmentioning

confidence: 99%

An allosteric hot spot in the tandem-SH2 domain of ZAP-70 regulates T-cell signaling

Gangopadhyay

Manna

Roy

et al. 2019

Preprint

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22T-cell receptor (TCR) signaling is initiated by recruiting ZAP-70 to the cytosolic part of TCR. ZAP-70, a 23 non-receptor tyrosine kinase, is composed of an N-terminal tandem SH2 (tSH2) domain connected to 24 the C-terminal kinase domain. The ZAP-70 is recruited to the membrane through binding of tSH2 25 domain and the doubly-phosphorylated ITAM motifs of CD3 chains in the TCR complex. Our results 26show that the tSH2 domain undergoes a biphasic structural transition while binding to the doubly-27 phosphorylated ITAM-ζ1 peptide. The C-terminal SH2 domain binds first to the phosphotyrosine 28 residue of ITAM peptide to form an encounter complex leading to subsequent binding of second 29 phosphotyrosine residue to the N-SH2 domain. We decipher a network of non-covalent interactions that 30 allosterically couple the two SH2 domains during binding to doubly-phosphorylated ITAMs. Mutation in 31 the allosteric network residues, for example, W165C, uncouples the formation of encounter complex to 32 the subsequent ITAM binding thus explaining the altered recruitment of ZAP-70 to the plasma 33 membrane causing autoimmune arthritis in mice. The proposed mechanism of allosteric coupling is 34 unique to ZAP-70, which is fundamentally different from Syk, a close homolog of ZAP-70 expressed in 35 B-cells. 36Significance 37 38 T-cell and B-cell signaling is initiated by the same family of non-receptor tyrosine kinases, ZAP-70 and 39 Syk, respectively. ZAP-70 and Syk share homologous sequence and similar structural architecture, yet 40 the two kinases differ in their mode of ligand recognition. ZAP-70 binds cooperatively to its ligand, 41whereas Syk binds uncooperatively. Spontaneous mutation (W165C) in the regulatory module of ZAP-42 70 impairs T-cell signaling causes autoimmune arthritis in SKG mice, the mechanism of which is 43 unknown. We showed that ZAP-70 regulatory module undergoes a biphasic structural transition while 44 binding to its ligand, which is fundamentally different from Syk. We presented a molecular mechanism 45 of cooperativity in ZAP-70 regulatory module that explains altered ligand binding by ZAP-70 mutant 46 found in SKG mice. 47 48 49 50 51The zeta-chain-associated protein tyrosine kinase, ZAP-70, is a non-receptor tyrosine kinase 52 crucial for T-cell signaling, development, activation, and proliferation(1-4). T-cell signaling is 53 commenced by the recruitment of two protein tyrosine kinase, Src family kinase Lck and ZAP-70, to the 54 activated molecular complex of T-cell antigen receptor (TCR)(5, 6). Lck, phosphorylate several tyrosine 55 residues of the immuno-receptor tyrosine-based activation motifs (ITAM) on the intracellular segment 56 of CD3 heterodimer (made up of d, g, and e) and ζ homodimer associated with the TCR(5, 7-10). ZAP-57 70 is spontaneously recruited to the membrane by binding to the doubly-phosphorylated ITAM (ITAM-58 Y2P) motifs (11)(12)(13)(14). Recruitment of ZAP-70 allows phosphorylation of scaffold proteins that initiates a 59 cascade of downstream biological events (15, 16)...

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ZAP-70 in Signaling, Biology, and Disease

Cited by 117 publications

References 184 publications

Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells

Intrathymic adeno-associated virus gene transfer rapidly restores thymic function and long-term persistence of gene-corrected T cells

T cell receptor-dependent S-acylation of ZAP-70 controls activation of T cells

An allosteric hot spot in the tandem-SH2 domain of ZAP-70 regulates T-cell signaling

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