A Phosphosite within the SH2 Domain of Lck Regulates Its Activation by CD45

Mol Cell. 2017 Aug 3;67(3):498-511.e6. doi: 10.1016/j.molcel.2017.06.024. Epub 2017 Jul 20.

Abstract

The Src Family kinase Lck sets a critical threshold for T cell activation because it phosphorylates the TCR complex and the Zap70 kinase. How a T cell controls the abundance of active Lck molecules remains poorly understood. We have identified an unappreciated role for a phosphosite, Y192, within the Lck SH2 domain that profoundly affects the amount of active Lck in cells. Notably, mutation of Y192 blocks critical TCR-proximal signaling events and impairs thymocyte development in retrogenic mice. We determined that these defects are caused by hyperphosphorylation of the inhibitory C-terminal tail of Lck. Our findings reveal that modification of Y192 inhibits the ability of CD45 to associate with Lck in cells and dephosphorylate the C-terminal tail of Lck, which prevents its adoption of an active open conformation. These results suggest a negative feedback loop that responds to signaling events that tune active Lck amounts and TCR sensitivity.

Keywords: CD45; Lck; SH2 domain; Src family kinase; T cell antigen receptor; T cell signaling; TCR; kinase; phosphatase; phosphorylation.

MeSH terms

  • Animals
  • Enzyme Activation
  • Genotype
  • HEK293 Cells
  • Humans
  • Jurkat Cells
  • Leukocyte Common Antigens / chemistry
  • Leukocyte Common Antigens / genetics
  • Leukocyte Common Antigens / metabolism*
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / chemistry
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / deficiency
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / genetics
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / metabolism*
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Models, Molecular
  • Mutation
  • Phenotype
  • Phosphorylation
  • Protein Binding
  • Proto-Oncogene Proteins c-fyn / genetics
  • Proto-Oncogene Proteins c-fyn / metabolism
  • Receptors, Antigen, T-Cell / metabolism
  • Signal Transduction
  • Thymocytes / enzymology*
  • Thymocytes / immunology
  • Time Factors
  • Transfection
  • src Homology Domains*

Substances

  • Receptors, Antigen, T-Cell
  • Fyn protein, mouse
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • Proto-Oncogene Proteins c-fyn
  • Leukocyte Common Antigens
  • PTPRC protein, human
  • Ptprc protein, mouse