The adaptor protein Lad associates with the G protein beta subunit and mediates chemokine-dependent T-cell migration

Blood. 2007 Jun 15;109(12):5122-8. doi: 10.1182/blood-2005-10-061838. Epub 2007 Feb 27.

Abstract

Lck-interacting adaptor protein/Rlk/Itk-binding protein (Lad/RIBP) was previously identified as an adaptor protein involved in TCR-mediated T-cell activation. To elucidate the functions of Lad further, we here performed yeast 2-hybrid screening using Lad as bait and discovered that the G protein beta subunit (G beta) is a Lad-binding partner. Since the most well-known G protein-coupled receptor in T cells is the chemokine receptor, we investigated whether Lad is involved in chemokine signaling. We found that, upon chemokine treatment, Lad associated with G beta in Jurkat T cells. Furthermore, ectopic expression of dominant-negative Lad or the reduction of endogenous Lad expression by siRNA impaired the chemokine-induced migration of T cells, indicating that Lad is required for chemokine-induced T-cell migration. Subsequent investigation of the signaling pathways revealed that, in response to chemokine, Lad associated with the tyrosine kinases Lck and Zap-70 and that Lad was essential for the activation of Zap-70. Moreover, Lad was required for the chemokine-dependent tyrosine phosphorylation of focal adhesion molecules that included Pyk2 and paxillin. Taken together, these data show that, upon chemokine stimulation, Lad acts as an adaptor protein that links the G protein beta subunit to the tyrosine kinases Lck and Zap-70, thereby mediating T-cell migration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Adaptor Proteins, Signal Transducing / physiology*
  • Chemokines / pharmacology*
  • Chemotaxis, Leukocyte*
  • GTP-Binding Protein beta Subunits / metabolism*
  • Humans
  • Jurkat Cells
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / metabolism
  • Protein Binding / drug effects
  • Protein-Tyrosine Kinases / metabolism
  • Signal Transduction
  • T-Lymphocytes / cytology
  • ZAP-70 Protein-Tyrosine Kinase / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Chemokines
  • GTP-Binding Protein beta Subunits
  • SH2D2A protein, human
  • Protein-Tyrosine Kinases
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • ZAP-70 Protein-Tyrosine Kinase