The Soluble Heparin-Binding EGF-Like Growth Factor Stimulates EGF Receptor Trafficking to the Nucleus

• Published in: PloS one 2015-05, Vol.10 (5), p.e0127887-e0127887
• Main Authors: Korotkevych, Nataliia V, Labyntsev, Andrii Ju, Kolybo, Denis V, Komisarenko, Serhiy V
• Format: Article
• Language: English
• Subjects: Anticoagulants; Binding; Biochemistry; Cancer; Cell Line; Cell Nucleus - drug effects; Cell Nucleus - metabolism; Chemical properties; Chromatin; Cloning; Confocal microscopy; Coordination compounds; Cyclin D1; Cyclin D1 - metabolism; Data processing; Deoxyribonucleic acid; DNA; Enzyme inhibitors; Epidermal growth factor; Epidermal growth factor receptors; Epidermal growth factors; Genes; Heparin; Heparin-binding EGF-like Growth Factor - metabolism; Humans; Immunology; Immunoprecipitation; Importation; Kinases; Ligands; Localization; Microscopy; Nuclear transport; Nuclei (cytology); Phosphorylation; Plasmids; Promoter Regions, Genetic - drug effects; Protein Kinase Inhibitors - pharmacology; Protein transport; Protein Transport - drug effects; Proteins; Quinazolines - pharmacology; Receptor, Epidermal Growth Factor - metabolism; Transcription; Translocation; Tumors; Tyrosine; Tyrphostins - pharmacology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0127887

Most ligands of epidermal growth factor receptor (EGFR) have the ability to induce EGFR translocation into the nucleus, where EGFR acts as an important transcriptional regulator. Soluble form of heparin-binding EGF-like growth factor (sHB-EGF) is one of the ligands for EGFR in many cell types; however, there is no evidence for the ability of sHB-EGF to induce EGFR nuclear importation. Here, we demonstrated that treatment of A431 cells with sHB-EGF resulted in nuclear localization of EGFR and such translocation occurs via retrograde pathway. It was shown by confocal microscopy and co-immunoprecipitation assay that the translocation complex consisted of both ligand and receptor. The chromatin immunoprecipitation assay showed the association of sHB-EGF-EGFR complex with promoter region of cyclin D1 in the cell nucleus and this association was prevented by application of EGFR kinase inhibitor AG-1478. The obtained data suggest that sHB-EGF acts similarly to other EGFR ligands and is capable to induce EGFR nuclear translocation as a part of ligand-receptor complex in a tyrosine phosphorylation-dependent manner.