Anti-oncogenic activity of signalling-defective epidermal growth factor receptor mutants.
Overexpression and autocrine activation of the epidermal growth factor receptor (EGF-R) cause transformation of cultured cells and correlate with tumor progression in cancer patients. Dimerization and transphosphorylation are crucial events in the process by which receptors with tyrosine kinase activity generate normal and transforming cellular signals. Interruption of this process by inactive receptor mutants offers the potential to inhibit ligand-induced cellular responses. Using recombinant retroviruses, we have examined the effects of signalling-incompetent EGF-R mutants on the growth-promoting and transforming potential of ligand-activated, overexpressed wild-type EGF-R and the v-erbB oncogene product. Expression of a soluble extracellular EGF-R domain had little if any effect on the growth and transformation of NIH 3T3 cells by either tyrosine kinase. However, both a kinase-negative EGF-R point mutant (HERK721A) and an EGF-R lacking 533 C-terminal amino acids efficiently inhibited wild-type EGF-R-mediated, de novo DNA synthesis and cell transformation in a dose-dependent manner. Furthermore, coexpression with the v-erbBES4 oncogene product in NIH 3T3 cells resulted in transphosphorylation of the HERK721A mutant receptor and reduced soft-agar colony growth but had no effect in a focus formation assay. These results demonstrate that signalling-defective receptor tyrosine kinase mutants differentially interfere with oncogenic signals generated by either overexpressed EGF-R or the retroviral v-erbBES4 oncogene product.
Department of Molecular Biology, Max-Planck-Institut für Biochemie, Martinsried, Germany.
N
N
Redemann
B
B
Holzmann
T
T
von Rüden
E F
EF
Wagner
J
J
Schlessinger
A
A
Ullrich
D016475:3T3 Cells; D000818:Animals; D004252:DNA Mutational Analysis; D004274:DNA, Recombinant / Q000235:genetics; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D009154:Mutation / Q000235:genetics; D018739:Oncogene Proteins v-erbB; D010766:Phosphorylation; D012190:Retroviridae / Q000235:genetics; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
D004274:DNA, Recombinant; D018739:Oncogene Proteins v-erbB; D015682:Retroviridae Proteins, Oncogenic; D004815:Epidermal Growth Factor; D066246:ErbB Receptors