Functional nuclear epidermal growth factor receptors in human choriocarcinoma JEG-3 cells and normal human placenta.
Immunocytochemistry with a monoclonal antiepidermal growth factor (anti-EGF) receptor antibody directed against the extracellular domain which can inhibit ligand binding to the receptors showed that nuclei of choriocarcinoma JEG-3 cells and normal placental trophoblasts were distinctly immunostained for EGF receptors. This finding led us to investigate the structure and function of nuclear EGF receptors. Western immunoblotting revealed that cell membranes, isolated intact pure nuclei, and nuclear membranes contain a 170-kilodalton EGF receptor protein. Covalent receptor cross-linking demonstrated that the 170-kilodalton receptor protein in nuclei and nuclear membranes can bind [125I]EGF just as in cell membranes, and that this binding is inhibited by excess unlabeled EGF. As in cell membranes, the addition of EGF resulted in an increased receptor autophosphorylation in the nuclei and nuclear membranes. In addition, the activated receptor kinase stimulated, and in some cases inhibited, tyrosine phosphorylation of a number of lower molecular size proteins, especially in nuclei and nuclear membranes. Although the identity of these proteins is not known, none of them could bind [125I]EGF. The addition of EGF to isolated nuclei resulted in a time-dependent specific transcriptional inhibition of hCG/LH receptor gene. In summary, our data demonstrating the presence of functional nuclear EGF receptors are novel, potentially important, and go against the traditional concepts of growth factors action. The nuclear receptors have the capacity to transduce signals from EGF and may mediate intracrine and paracrine actions of EGF in the regulation of trophoblast functions.
Cao
H
H
Department of Obstetrics and Gynecology, University of Louisville School of Medicine, Kentucky 40292, USA.
Lei
Z M
ZM
Bian
L
L
Rao
C V
CV
D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus / Q000737:chemistry; D002822:Choriocarcinoma / Q000737:chemistry* / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology; D066246:ErbB Receptors / Q000032:analysis* / Q000737:chemistry / Q000502:physiology; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007163:Immunosorbent Techniques; D016253:Microscopy, Immunoelectron; D009685:Nuclear Envelope / Q000737:chemistry; D010766:Phosphorylation; D019000:Phosphotyrosine; D010920:Placenta / Q000737:chemistry* / Q000648:ultrastructure; D011247:Pregnancy; D011974:Receptors, LH / Q000235:genetics; D015398:Signal Transduction; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D011974:Receptors, LH; D019000:Phosphotyrosine; D014443:Tyrosine; D004815:Epidermal Growth Factor; D066246:ErbB Receptors