Inhibition of the kinase Csk in thymocytes reveals a requirement for actin remodeling in the initiation of full TCR signaling

Tan, Ying Xim; Manz, Boryana N.; Freedman, Tanya S.; Zhang, Chao; Shokat, Kevan M.; Weiss, Arthur

doi:10.1038/ni.2772

Cited by 86 publications

(111 citation statements)

References 54 publications

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“…With the exception of PKA and CSK, the expression of these kinases is predominantly restricted to immune cells, suggesting selective roles in immune modulation. Accordingly, genetically engineered mouse models enabling the abrogation of PKA, CSK, MAP4K1, or DGKξ functions exhibit hallmarks of excessive immune activation and/or TCR hypersensitivity (128,(130)(131)(132) -/-mice, the role of these kinases in tumor immune surveillance has already been established, confirming the viability of these approaches for restoring antitumor immunity (131,133). In contrast, ubiquitous expression and pleiotropic functions of CSK and PKA may limit the viability of these targets in this context.…”

Section: Future Directionsmentioning

confidence: 87%

Targeting cancer with kinase inhibitors

Größ¹,

Rahal²,

Stransky³

et al. 2015

J. Clin. Invest.

385

289

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Section: Future Directionsmentioning

confidence: 87%

Targeting cancer with kinase inhibitors

Größ¹,

Rahal²,

Stransky³

et al. 2015

J. Clin. Invest.

385

289

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“…Prominent among these pathways are those that involve remodeling the actin cytoskeleton and the PI3K pathway. As a preliminary effort to explore this dataset, we focused on the actin cytoskeleton because our recent studies (32) and those of others (29) suggested an important role for CD28 in regulating the actin cytoskeleton. The genetic study of Liang et al (29), which identified an important role of Rlptr, led us to focus on CapZIP.…”

Section: Discussionmentioning

confidence: 99%

Combinatorial proteomic analysis of intercellular signaling applied to the CD28 T-cell costimulatory receptor

Tian

Wang²,

Gish

et al. 2015

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

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Systematic characterization of intercellular signaling approximating the physiological conditions of stimulation that involve direct cell-cell contact is challenging. We describe a proteomic strategy to analyze physiological signaling mediated by the T-cell costimulatory receptor CD28. We identified signaling pathways activated by CD28 during direct cell-cell contact by global analysis of protein phosphorylation. To define immediate CD28 targets, we used phosphorylated forms of the CD28 cytoplasmic region to obtain the CD28 interactome. The interaction profiles of selected CD28-interacting proteins were further characterized in vivo for amplifying the CD28 interactome. The combination of the global phosphorylation and interactome analyses revealed broad regulation of CD28 and its interactome by phosphorylation. Among the cellular phosphoproteins influenced by CD28 signaling, CapZ-interacting protein (CapZIP), a regulator of the actin cytoskeleton, was implicated by functional studies. The combinatorial approach applied herein is widely applicable for characterizing signaling networks associated with membrane receptors with short cytoplasmic tails.uring some forms of intercellular communication, soluble growth factors are secreted by one cell type and bind specific transmembrane receptor tyrosine kinases (RTKs) present on neighboring cells. Activated RTKs then undergo autophosphorylation and also phosphorylate associated scaffold proteins, thereby creating docking sites for effector proteins with phosphotyrosine (pTyr)-recognition modules such as Src homology 2 (SH2) and pTyr-binding (PTB) domains (1). These immediate targets of growth-factor receptors control a variety of intracellular responses, typified by the activation of serine/ threonine protein kinases such as those involved in the ERK MAP kinase pathway. Growth-factor signaling therefore leads to extensive changes in both tyrosine and serine/threonine phosphorylation (2).The alterations in protein-protein interactions and posttranslational modifications elicited by transmembrane receptors can in principle be characterized by mass spectrometry (MS)-based proteomics (3-6). However, under circumstances that involve complex cell-cell interactions, identifying physiologically relevant signals using MS-based proteomics can be challenging. For example, obtaining a sufficiently robust response for MS analysis has often required the use of cells that overexpress a receptor of interest and are subjected to a nonphysiological stimulus. For this reason, experiments have often focused on specific, often stereotypic, aspects of the cellular response, rather than taking a more global approach. Moreover, cell-cell interactions are often typified by the interaction of multiple transmembrane receptor-ligand pairs, frequently involving membrane proteins that are not receptor kinases. Short-peptide motifs from such receptors that serve as potential ligands for downstream targets can be used as affinity probes for binding partners, but these experiments suffer from high fals...

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“…In the ground state, the levels of active LCK are set to a point ensuring that maximal CD3 ITAM and ZAP-70 phosphorylation only occurs upon engagement of the TCR with agonist pMHC ligands. Consistent with the existence of such basal PTK-PTPase balance, its alteration in favor of PTK via genetic or pharmacological interventions can increase TCR sensitivity (9) and even trigger T cell activation, irrespective of TCR-pMHC engagement (5,10).…”

Section: T He Recognition Of Peptide-mhc (Pmhc) Ligands By T Cells Anmentioning

confidence: 82%

“…For instance, in response to very weak interactions with self-pMHC, the TCR signaling pathway found in naive T cells, rather than being silent, delivers low-intensity signals that imprint on naive T cells a heightened reactivity to foreign pMHC (2). A network of PTKs, protein tyrosine phosphatases (PTPases), and transmembrane adaptors dynamically control the activity of LCK in the ground state (3)(4)(5)(6)(7)(8). In the ground state, the levels of active LCK are set to a point ensuring that maximal CD3 ITAM and ZAP-70 phosphorylation only occurs upon engagement of the TCR with agonist pMHC ligands.…”

Section: T He Recognition Of Peptide-mhc (Pmhc) Ligands By T Cells Anmentioning

confidence: 99%

Revisiting the Timing of Action of the PAG Adaptor Using Quantitative Proteomics Analysis of Primary T Cells

Reginald

Chaoui

Roncagalli

et al. 2015

The Journal of Immunology

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The protein tyrosine kinase LCK plays a key role in TCR signaling, and its activity is dynamically controlled by the tyrosine kinase C-terminal Src kinase (CSK) and the tyrosine phosphatase CD45. CSK is brought in contiguity to LCK via binding to a transmembrane adaptor known as phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG). The lack of a blatant phenotype in PAG-deficient mice has impeded our understanding of the mechanisms through which PAG exerts its negative-regulatory role in TCR signaling. We used quantitative mass spectrometry and both thymocytes and CD4+ T cells from mice in which a tag for affinity purification was knocked in the gene coding for PAG to determine the composition and dynamics of the multiprotein complexes that are found around PAG over 5 min of activation. Most of the high-confidence interactions that we observed were previously unknown. Using phosphoproteomic analysis, PAG showed low levels of tyrosine phosphorylation in resting primary mouse CD4+ T cells; the levels of tyrosine phosphorylation increased and reached a maximum 2 min after stimulation. Analysis of the dynamics of association of the protein tyrosine phosphatase PTPN22 and lipid phosphatase SHIP-1 with PAG following T cell activation suggests that both cooperate with CSK to terminate T cell activation. Our findings provide a model of the role for PAG in mouse primary CD4+ T cells that is consistent with recent phosphoproteomic studies of the Jurkat T cell line but difficult to reconcile with former biochemical studies indicating that PAG is constitutively phosphorylated in resting T cells and rapidly dephosphorylated once the TCR is engaged.

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Inhibition of the kinase Csk in thymocytes reveals a requirement for actin remodeling in the initiation of full TCR signaling

Cited by 86 publications

References 54 publications

Targeting cancer with kinase inhibitors

Targeting cancer with kinase inhibitors

Combinatorial proteomic analysis of intercellular signaling applied to the CD28 T-cell costimulatory receptor

Revisiting the Timing of Action of the PAG Adaptor Using Quantitative Proteomics Analysis of Primary T Cells

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