The RING finger of c-Cbl mediates desensitization of the epidermal growth factor receptor.
Ligand-induced activation of surface receptors, including the epidermal growth factor receptor (EGFR), is followed by a desensitization process involving endocytosis and receptor degradation. c-Cbl, a tyrosine phosphorylation substrate shared by several signaling pathways, accelerates desensitization by recruiting EGFR and increasing receptor polyubiquitination. Here we demonstrate that the RING type zinc finger of c-Cbl is essential for ubiquitination and subsequent desensitization of EGFR. Mutagenesis of a single cysteine residue impaired the ability of c-Cbl to enhance both down-regulation and ubiquitination of EGFR in living cells, although the mutant retained binding to the activated receptor. Consequently, the mutant form of c-Cbl acquired a dominant inhibitory function and lost the ability to inhibit signaling downstream to EGFR. In vitro reconstitution of EGFR ubiquitination implies that the RING finger plays an essential direct role in ubiquitin ligation. Our results attribute to the RING finger of c-Cbl a causative role in endocytic sorting of EGFR and desensitization of signal transduction.
Waterman
H
H
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel.
Levkowitz
G
G
Alroy
I
I
Yarden
Y
Y
D015536:Down-Regulation; D004705:Endocytosis; D066246:ErbB Receptors / Q000378:metabolism*; D016296:Mutagenesis; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D050721:Proto-Oncogene Proteins c-cbl; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000378:metabolism; D016335:Zinc Fingers
D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D014452:Ubiquitins; D050721:Proto-Oncogene Proteins c-cbl; D044767:Ubiquitin-Protein Ligases; D066246:ErbB Receptors