HS1 Functions as an Essential Actin-Regulatory Adaptor Protein at the Immune Synapse

Gomez, Timothy S.; McCarney, Sean; Carrizosa, Esteban; Labno, Christine M.; Comiskey, Erin O.; Nolz, Jeffrey C.; Zhu, Peimin; Freedman, Bruce D.; Clark, Marcus R.; Rawlings, David J.; Billadeau, Daniel D.; Burkhardt, Janis K.

doi:10.1016/j.immuni.2006.03.022

Cited by 187 publications

(252 citation statements)

References 46 publications

(74 reference statements)

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“…In detail, the HS1 sites Y 378 and Y 397 are phosphorylated by Syk in vitro [36] and presumably by ZAP70 upon TCR ligation [27]. Besides these tyrosine motifs, in vitro phosphorylation of Y 222 (YKKT) [36][37][38] and Y 198 (YGIQ) [39] was documented.…”

Section: Discussionmentioning

confidence: 94%

“…In a first series of experiments we verified this interaction and show that both Nck1 and Nck2 associate with HS1. [24,27]. Of note, Nck also plays an important role in linking TCR signaling to the actin cytoskeleton within the immunological synapse.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, HS1 was shown to be phosphorylated (most likely by ZAP70) at Y 378 and Y 397 upon TCR ligation. Then Itk recruits phosphorylated HS1 to the IS, where HS1 stabilizes F-actin and modulates Vav1 and PLCγ1 localization [24,27]. Of note, Nck also plays an important role in linking TCR signaling to the actin cytoskeleton within the immunological synapse.…”

Section: Discussionmentioning

confidence: 99%

“…An SH3 domain forms the C-terminal part of the molecule [3,22]. Moreover, HS1 contains several tyrosine motifs and is a substrate of Src and Syk kinases in several hematopoietic cell types [23][24][25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, a role for HS1 in T-cell activation has been established where HS1 is required for sustained actin responses at the immunological synapse, Ca 2+ signaling and IL2 gene expression [27]. In this scenario, HS1 is phosphorylated upon TCR ligation, presumably by ZAP70.…”

Section: Introductionmentioning

confidence: 99%

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SDF1α‐induced interaction of the adapter proteins Nck and HS1 facilitates actin polymerization and migration in T cells

2014

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Noncatalytic region of tyrosine kinase (Nck) is an adapter protein that comprises one SH2 (Src homology) domain and three SH3 domains. Nck links receptors and receptorassociated tyrosine kinases or adapter proteins to proteins that regulate the actin cytoskeleton. Whereas the SH2 domain binds to phosphorylated receptors or associated phosphoproteins, individual interactions of the SH3 domains with proline-based recognition motifs result in the formation of larger protein complexes. In T cells, changes in cell polarity and morphology during T-cell activation and effector function require the T-cell receptor-mediated recruitment and activation of actin-regulatory proteins to initiate cytoskeletal reorganization at the immunological synapse. We previously identified the adapter protein HS1 as a putative Nck-interacting protein. We now demonstrate that the SH2 domain of Nck specifically interacts with HS1 upon phosphorylation of its tyrosine residue 378. We report that in human T cells, ligation of the chemokine receptor CXCR4 by stromal cell-derived factor 1α (SDF1α) induces a rapid and transient phosphorylation of tyrosine 378 of HS1 resulting in an increased association with Nck. Consequently, siRNA-mediated downregulation of HS1 and/or Nck impairs SDF1α-induced actin polymerization and T-cell migration.Keywords: CXCR4 r HS1 r Nck r SDF1α r T cells IntroductionT cells govern adaptive immunity by cytokine secretion or cytotoxic effector function. After development in the thymus, naive T lymphocytes constantly recirculate between the blood stream and lymphoid tissues. At the molecular level, this homing and migration processes rely on complex interactions of selected chemokines with their receptors on respective T cells and of T-cell adhesion molecules binding to their ligands on endothelial cells or the extracellular matrix. Individual interactions elicit specific signaling pathways that induce dynamic cytoskeletal rearrangements, which enable lymphocytes to adhere to or pass a certain substrate or to transmigrate through a given endothelium. Morphologically, circular floating T cells adopt a migratory phenotype characterized Correspondence: Dr. Marcus Lettau e-mail: lettau@immunologie.uni-kiel.de by a leading edge at the front and an uropod at the rear. When T cells encounter their antigen on an antigen-presenting cell in the lymph nodes or spleen, migration is halted and the cells rapidly polarize toward the intercellular contact area. The subsequent formation of the so-called immunological synapse (IS) is again accompanied by complex cytoskeletal changes and comprises the structural basis for T-cell activation. This primary antigenic stimulation ultimately results in cell-cycle progression, clonal expansion, and differentiation to T-effector cells. The activated T cells regain migratory potential, leave the lymphoid tissue, and search the periphery for infected or transformed cells displaying their cognate antigen. Upon antigen encounter, the T cells adhere to their target cell and polarize toward the intercell...

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

SDF1α‐induced interaction of the adapter proteins Nck and HS1 facilitates actin polymerization and migration in T cells

2014

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Artificial Biosystem for Modulation of Interactions between Antigen‐Presenting Cells and T Cells

et al. 2020

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T cell activation is triggered by signal molecules on the surface of antigen‐presenting cells (APC) and subsequent exertion of cellular forces. Deciphering the biomechanical and biochemical signals in this complex process is of interest and will contribute to an improvement in immunotherapy strategies. To address underlying questions, coculture and biomimetic models are established. Mature dendritic cells (mDC) are first treated with cytochalasin B (CytoB), a cytoskeletal disruption agent known to lower apparent cellular stiffness and reduction in T cell proliferation is observed. It is attempted to mimic mDC and T cell interactions using polyacrylamide (PA) gels with defined stiffness corresponding to mDC (0.2–25 kPa). Different ratios of anti‐CD3 (aCD3) and anti‐CD28 (aCD28) antibodies are immobilized onto PA gels. The results show T cell proliferation is triggered by both aCD3 and aCD28 in a stiffness‐dependent manner. Cells cultured on aCD3 immobilized on gels has significantly enhanced proliferation and IL‐2 secretion, compared to aCD28. Furthermore, ZAP70 phosphorylation is enhanced in stiffer substrate a in a aCD3‐dependent manner. The biosystem provides an approach to study the reduction of T cell proliferation observed on CytoB‐treated mDC. Overall, the biosystem allows distinguishing the impact of biophysical and biochemical signals of APC and T cell interactions in vitro.

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Differential recognition of syk‐binding sites by each of the two phosphotyrosine‐binding pockets of the Vav SH2 domain

et al. 2013

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The association of Syk, a central tyrosine kinase in B cell signaling, with Vav SH2 domain is controlled by phosphorylation of two closely spaced tyrosines in Syk linker B: Y342 and Y346. Previous studies established both singly phosphorylated and doubly phosphorylated forms play a role in signaling. The structure of the doubly phosphorylated form identified a new recognition of phosphotyrosine whereby two phosphotyrosines bind simultaneously to the Vav SH2 domain, one in the canonical pTyr pocket and one in the specificity pocket on the opposite side of the central β-sheet. It is unknown if the specificity pocket can bind phosphotyrosine independent of phosphotyrosine binding the pTyr pocket. To address this gap in knowledge, we determined the structure of the complex between Vav1 SH2 and a peptide (SykLB-YpY) modeling the singly phosphorylated-Y346 form of Syk with unphosphorylated Y342. The NMR data conclusively establish that recognition of phosphotyrosine is swapped between the two pockets; phosphorylated pY346 binds the specificity pocket of Vav1 SH2, and unphosphorylated Y342 occupies what is normally the pTyr binding pocket. Nearly identical changes in chemical shifts occurred upon binding all three forms of singly and doubly phosphorylated peptides; however somewhat smaller shift perturbations for SykLB-YpY from residues in regions of high internal mobility suggest that internal motions are coupled to binding affinity. The differential recognition that includes this swapped binding of phosphotyrosine to the specificity pocket of Vav SH2 increases the repertoire of possible phosphotyrosine binding by SH2 domains in regulating protein-protein interactions in cellular signaling.

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HS1 Functions as an Essential Actin-Regulatory Adaptor Protein at the Immune Synapse

Cited by 187 publications

References 46 publications

SDF1α‐induced interaction of the adapter proteins Nck and HS1 facilitates actin polymerization and migration in T cells

SDF1α‐induced interaction of the adapter proteins Nck and HS1 facilitates actin polymerization and migration in T cells

Artificial Biosystem for Modulation of Interactions between Antigen‐Presenting Cells and T Cells

Differential recognition of syk‐binding sites by each of the two phosphotyrosine‐binding pockets of the Vav SH2 domain

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