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The Journal of Immunology, 2002, 169: 292-300.
Copyright © 2002 by The American Association of Immunologists

TCR Engagement Induces Proline-Rich Tyrosine Kinase-2 (Pyk2) Translocation to the T Cell-APC Interface Independently of Pyk2 Activity and in an Immunoreceptor Tyrosine-Based Activation Motif-Mediated Fashion1

David Sancho*, María C. Montoya*, Alicia Monjas{dagger}, Mónica Gordón-Alonso*, Takuya Katagiri{ddagger}, Diana Gil{dagger}, Reyes Tejedor*, Balbino Alarcón{dagger} and Francisco Sánchez-Madrid2,*

* Servicio de Inmunología, Hospital de la Princesa, and {dagger} Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid, Madrid, Spain; and {ddagger} Division of Biochemistry and Cellular Biology, National Institute of Neuroscience, Tokyo, Japan

The relocation of kinases in T lymphocytes during their cognate interaction with APCs is essential for lymphocyte activation. We found that the proline-rich tyrosine kinase-2 (Pyk2) is rapidly translocated to the T cell-APC contact area upon T cell-specific recognition of superantigen-pulsed APCs. Stimulation with anti-CD3-coated latex microspheres was sufficient for Pyk2 reorientation, and the coengagement of CD28 boosted Pyk2 redistribution. Nevertheless, Pyk2 translocation did not result in its recruitment to lipid rafts. Two results support that Pyk2 translocation was independent of its kinase activity. First, Lck activity was required for TCR-induced Pyk2 translocation, but not for TCR-induced Pyk2 activation. Second, a kinase-dead Pyk2 mutant was equally translocated upon TCR triggering. In addition, Lck activity alone was insufficient to induce Pyk2 reorientation and activation, requiring the presence of at least one intact immunoreceptor tyrosine-based activation motif (ITAM). Despite the dependence on functional Lck and on phosphorylated ITAM for Pyk2 translocation, the ITAM-binding tyrosine kinase {zeta}-associated protein 70 (ZAP-70) was not essential. All these data suggest that, by translocating to the vicinity of the immune synapse, Pyk2 could play an essential role in T cell activation and polarized secretion of cytokines.




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