Functional analysis of LAT in TCR-mediated signaling pathways using a LAT-deficient Jurkat cell line

Int Immunol. 1999 Jun;11(6):943-50. doi: 10.1093/intimm/11.6.943.


The adaptor molecule LAT (linker for activation of T cells) is a palmitoylated integral membrane protein that localizes to the glycolipid-enriched microdomains in the plasma membrane. Upon TCR engagement, LAT becomes phosphorylated on multiple tyrosine residues and then binds several critical signaling molecules. Here, we describe the generation and characterization of a LAT-deficient cell line. Using this cell line, we demonstrate that LAT is required for TCR-mediated Ca2+ mobilization and optimal tyrosine phosphorylation of phospholipase C-gamma1, Vav and SLP-76. LAT is also required for Erk activation, CD69 up-regulation, and AP- and NFAT-mediated gene transcription. We also demonstrate, by reconstituting this cell line with LAT mutants, that the LAT transmembrane domain and palmitoylation at Cys26, but not Cys29, are required for LAT function and TCR signaling. These studies provide further evidence for the crucial role of the LAT molecule, and demonstrate the use of a LAT-deficient cell line for the analysis of LAT structure and function.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing*
  • Antigens, CD / biosynthesis
  • Antigens, Differentiation, T-Lymphocyte / biosynthesis
  • Binding Sites
  • Calcium-Calmodulin-Dependent Protein Kinases / metabolism
  • Carrier Proteins / biosynthesis*
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism
  • Cell Separation
  • Clone Cells
  • Enzyme Activation / immunology
  • Humans
  • Isoenzymes / metabolism
  • Jurkat Cells / enzymology
  • Jurkat Cells / immunology*
  • Jurkat Cells / metabolism*
  • Lectins, C-Type
  • Membrane Proteins*
  • Mutation
  • Oncogene Proteins / metabolism
  • Palmitic Acid / metabolism
  • Phospholipase C gamma
  • Phosphoproteins / biosynthesis*
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Proto-Oncogene Proteins c-vav
  • Receptors, Antigen, T-Cell / physiology*
  • Signal Transduction / genetics
  • Signal Transduction / immunology*
  • Transcriptional Activation / immunology
  • Transfection
  • Type C Phospholipases / metabolism
  • Tyrosine / metabolism
  • Up-Regulation / immunology
  • src Homology Domains / immunology


  • Adaptor Proteins, Signal Transducing
  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD69 antigen
  • Carrier Proteins
  • Isoenzymes
  • LAT protein, human
  • Lectins, C-Type
  • Membrane Proteins
  • Oncogene Proteins
  • Phosphoproteins
  • Proto-Oncogene Proteins c-vav
  • Receptors, Antigen, T-Cell
  • SLP-76 signal Transducing adaptor proteins
  • VAV1 protein, human
  • Palmitic Acid
  • Tyrosine
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Type C Phospholipases
  • Phospholipase C gamma