Original Article

The Tyrosine Kinase PYK-2/RAFTK Regulates Natural Killer (NK) Cell Cytotoxic Response, and Is Translocated and Activated upon Specific Target Cell Recognition and Killing

Correspondence to: Francisco Sánchez-Madrid, C/Diego de León, 62, 28006 Madrid, Spain. Tel:34-91-5202370 Fax:34-91-5202374 E-mail:fsanchez{at}hlpr.insalud.es.

The compartmentalization of plasma membrane proteins has a key role in regulation of lymphocyte activation and development of immunity. We found that the proline-rich tyrosine kinase-2 (PYK-2/RAFTK) colocalized with the microtubule-organizing center (MTOC) at the trailing edge of migrating natural killer (NK) cells. When polyclonal NK cells bound to K562 targets, PYK-2 translocated to the area of NK–target cell interaction. The specificity of this process was assessed with NK cell clones bearing activatory or inhibitory forms of CD94/NKG2. The translocation of PYK-2, MTOC, and paxillin to the area of NK–target cell contact was regulated upon specific recognition of target cells through NK cell receptors, controlling target cell killing. Furthermore, parallel in vitro kinase assays showed that PYK-2 was activated in response to signals that specifically triggered its translocation and NK cell mediated cytotoxicity. The overexpression of both the wt and a dominant-negative mutant of PYK-2, but not ZAP-70 wt, prevented the specific translocation of the MTOC and paxillin, and blocked the cytotoxic response of NK cells. Our data indicate that subcellular compartmentalization of PYK-2 correlates with effective signal transduction. Furthermore, they also suggest an important role for PYK-2 on the assembly of the signaling complexes that regulate the cytotoxic response.

Key Words: CD94/NKG2, cytotoxicity, microtubule-organizing center, cytoskeletal proteins, HLA-E

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