Loss of c-Cbl RING finger function results in high-intensity TCR signaling and thymic deletion

EMBO J. 2005 Nov 2;24(21):3807-19. doi: 10.1038/sj.emboj.7600841. Epub 2005 Oct 6.

Abstract

Signaling from the T-cell receptor (TCR) in thymocytes is negatively regulated by the RING finger-type ubiquitin ligase c-Cbl. To further investigate this regulation, we generated mice with a loss-of-function mutation in the c-Cbl RING finger domain. These mice exhibit complete thymic deletion by young adulthood, which is not caused by a developmental block, lack of progenitors or peripheral T-cell activation. Rather, this phenotype correlates with greatly increased expression of the CD5 and CD69 activation markers and increased sensitivity to anti-CD3-induced cell death. Thymic loss contrasts the normal fate of the c-Cbl-/- thymus, even though thymocytes from both mutant mice show equivalent enhancement in proximal TCR signaling, Erk activation and calcium mobilization. Remarkably, only the RING finger mutant thymocytes show prominent TCR-directed activation of Akt. We show that the mutant c-Cbl protein itself is essential for activating this pathway by recruiting the p85 regulatory subunit of PI 3-kinase. This study provides a unique model for analyzing high-intensity TCR signals that cause thymocyte deletion and highlights multiple roles of c-Cbl in regulating this process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Antigens, CD / analysis
  • Antigens, Differentiation, T-Lymphocyte / analysis
  • Apoptosis
  • CD3 Complex / analysis
  • CD5 Antigens / analysis
  • Extracellular Signal-Regulated MAP Kinases / analysis
  • Lectins, C-Type
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / analysis
  • Mice
  • Mice, Transgenic
  • Mutation
  • Phosphatidylinositol 3-Kinases / metabolism
  • Protein Structure, Tertiary
  • Proto-Oncogene Proteins c-cbl / chemistry
  • Proto-Oncogene Proteins c-cbl / genetics
  • Proto-Oncogene Proteins c-cbl / physiology*
  • Receptors, Antigen, T-Cell / immunology
  • Receptors, Antigen, T-Cell / physiology*
  • Signal Transduction*
  • Thymus Gland / immunology*

Substances

  • Antigens, CD
  • Antigens, Differentiation, T-Lymphocyte
  • CD3 Complex
  • CD5 Antigens
  • CD69 antigen
  • Lectins, C-Type
  • Receptors, Antigen, T-Cell
  • Proto-Oncogene Proteins c-cbl
  • Phosphatidylinositol 3-Kinases
  • Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
  • Extracellular Signal-Regulated MAP Kinases
  • Cbl protein, mouse