Functional reconstitutional of the human epidermal growth factor receptor system in Xenopus oocytes.
We have expressed the human EGF receptor (hEGF-R) in Xenopus oocytes by injecting mRNA synthesized in vitro using SP6 vectors containing receptor cDNAs. Each oocyte could express over 1 x 10(10) receptors of a single affinity class and these were able to bind and rapidly internalize EGF. Occupancy resulted in receptor tyrosine autophosphorylation, downregulation, and release of intracellular calcium. Occupied receptors also rapidly induced meiotic maturation in stage VI oocytes. Receptors lacking tyrosine kinase activity bound EGF normally, but did not downregulate or induce any biological responses. The rate of oocyte maturation was proportional to hEGF-R occupancy and was significantly faster than progesterone-induced maturation at nanomolar EGF concentrations. Mutant hEGF-R truncated at residue 973 displayed identical phenotypes in both mammalian cells and oocytes in that they were defective in their ability to release intracellular calcium, undergo ligand induced internalization and receptor downregulation. However, these receptors were fully capable of inducing oocyte maturation. The remarkable retention of specific biological activities of different hEGF-R in the context of oocytes suggests that this receptor system interacts with generally available cellular components that have been conserved during evolution. In addition, it suggests that cell surface tyrosine kinase activity may play an important role in regulating resumption of the cell cycle.
Opresko
L K
LK
Department of Pathology, University of Utah Medical Center, Salt Lake City 84132.
Wiley
H S
HS
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics / Q000502:physiology*; D005822:Genetic Vectors; D006801:Humans; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008540:Meiosis; D008845:Microinjections; D016297:Mutagenesis, Site-Directed; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D011061:Poly A / Q000096:biosynthesis; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins; D014982:Xenopus laevis
D012333:RNA, Messenger; D011994:Recombinant Proteins; D011061:Poly A; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D016203:CDC2 Protein Kinase; D016200:Maturation-Promoting Factor