RANTES induces tyrosine kinase activity of stably complexed p125FAK and ZAP-70 in human T cells.

Abstract

SummaryThe chemokine RANTES is a chemoattractant and activating factor for T lymphocytes. Investigation of the signal transduction mechanisms induced by RANTES in T cells revealed tyrosine phosphorylation of multiple protein species with prominent bands at 70-85 and 120-130 kD. Immunoprecipitation and Western analyses revealed that a protein of 125 kD was identical to the focal adhesion kinase (FAK) pp125 FAK. R.ANTES stimulated phosphorylation of FAK as early as 30 seconds, and immunoblots using antiphosphotyrosine monoclonal antibodies revealed that there was consistent phosphorylation of a 68-70 kD species in the pp125 FAK immunoprecipitates. Immunoblotting and kinase assays showed this to be two separate proteins, the tyrosine kinase zeta-associated protein (ZAP) 70, and the focal adhesion protein paxillin. These results indicate a potentially important role for RANTES in the generation of T cell focal adhesions and subsequent cell activation via a molecular complex containing FAK, ZAP-70, and paxiltin.

Chemokines: attractive mediators of the immune response

“…Notably, CCL5 induced a similar protein phosphorylation profile to that observed following direct TCR activation, indicating that CCL5 may activate proximal TCR signaling events. In a subsequent study, Bacon et al [79] provided evidence that CCL5 treatment of T cells induces the phosphorylation and association of FAK, ZAP-70 and paxillin, as well as partial phosphorylation of the TCR-chain. Additionally, CCL4 can mediate a pattern of protein phosphorylation similar to direct CD3 stimulation in human thymocytes [85].…”

“…Since chemokines mediate cell migration, cellular polarization and activation, their ability to invoke FAK activation has received attention. CCL5 will induce the phosphorylation of FAK in human T cells [79] and its association with CCR5 [40]. Furthermore, CCL5 and CCL4 can induce the phosphorylation of Pyk2 in the human T cell line, DU6 [80] and Pyk2 activation by CCL4 has been shown to be dependent on CCR5 activation [81].…”

“…The implications are that Jaks may also be important for chemokine inducible cell migration, polarization, activation and differentiation. FAK may serve as a scaffold for the assembly of signaling intermediates activated by chemokines, since CCL5 treatment leads to the association of phosphorylated FAK with a critical tyrosine kinase in TCR activation, zeta-associated protein (ZAP)-70 [79]. Interestingly, both FAK and ZAP-70 phosphorylation are increased by their coassociation, indicating that each regulates the other's activation.…”

TNF-α-Induced Secretion of C-C Chemokines Modulates C-C Chemokine Receptor 5 Expression on Peripheral Blood Lymphocytes

“…Another report shows an enhancement of tyrosine phosphorylation of CCR5 and the association of Janus kinase 1/STAT5 with CCR5 after activation by RANTES (53). Furthermore, RANTES can lead to focal adhesion kinase association with CCR5 and Zap70; Lck and Pyk2 are activated after ligand binding (53)(54)(55)(56). How these signals participate in the internalization process or other signaling events is not yet established.…”

Augmentation of RANTES-Induced Extracellular Signal-Regulated Kinase Mediated Signaling and T Cell Adhesion by Elastase-Treated Fibronectin

“…These processes are associated with phosphatidylinositol 3 kinase signaling (13), a transient increase in cytosolic Ca 2ϩ concentration, and tyrosine phosphorylation of several proteins, such as FAK, ZAP-70, and paxillin. The latter molecules form a set of cytoskeletal-based elements implicated in the formation of focal adhesion contacts, cell adhesion, and activation (14). Recently, it was shown that stromal cell-derived factor (SDF)-1␣, a potent T cell chemoattractant (8,15), induces activation via another family of signaling moieties, the mitogen-activated protein (MAP) kinases (16).…”