The biological activity of the human epidermal growth factor receptor is positively regulated by its C-terminal tyrosines

Oncogene. 1991 May;6(5):825-32.


The epidermal growth factor receptor (EGF-R) C-terminus contains three conserved tyrosines (Y-1068, Y-1148, Y-1173) which are phosphorylated upon EGF activation. To clarify the functional role of these tyrosines, each has been mutated to phenylalanine and studied as single, double and triple mutants in the full length receptor. EGF-dependent transforming ability of the single point mutants is similar to that of the wild type, while that of double mutants is decreased and an even lower activity is present in the triple mutant. In each bioassay, including EGF-dependent focal transformation, growth in agar and growth in low serum, mutant receptors display a similar hierarchy of activity. The lower activity is intrinsic in the mutants since they are expressed at similar level as the wild type and bind EGF with similar affinity. Deletion mutants lacking the last 19 or 63 amino acids (Velu et al., 1989a) show a similar decline in biological activity when compared to the corresponding point mutants, although the reduction is more pronounced than with the point mutants. Deletion of the last 123 aa, which removes all three tyrosines (Dc123), results in a receptor that is almost inactive biologically. The EGF-R kinase activity is affected by tyrosine substitution since in vitro phosphorylation of exogenous substrates is reduced in the double and triple mutants. Autophosphorylation, in vivo and in vitro, is also reduced, but not totally abolished in the triple point mutant and Dc123 indicating the existence of other autophosphorylation sites. A new site of autophosphorylation is found in the Dc123 mutant. We conclude, therefore, that the tyrosines at the extreme C-terminus positively regulate the biological and transforming activity of the EGF-R, probably via autophosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cloning, Molecular
  • Epidermal Growth Factor / metabolism
  • ErbB Receptors / genetics*
  • ErbB Receptors / metabolism
  • Kinetics
  • Mice
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed*
  • Oligonucleotide Probes
  • Phosphopeptides / isolation & purification
  • Phosphorylation
  • Protein-Tyrosine Kinases / genetics*
  • Protein-Tyrosine Kinases / metabolism
  • Proto-Oncogenes*
  • Transfection*
  • Trypsin


  • Oligonucleotide Probes
  • Phosphopeptides
  • Epidermal Growth Factor
  • ErbB Receptors
  • Protein-Tyrosine Kinases
  • Trypsin