Regulation of the ErbB3 binding protein Ebp1 by protein kinase C.
Ebp1, a 47 kDa ubiquituously expressed protein, binds the ErbB3 receptor in human serum starved breast cancer cell lines and dissociates from ErbB3 on treatment with the ErbB3 ligand, Heregulin (HRG). However, the mechanism of Ebp1-ErbB3 association/dissociation is not understood. Since Ebp1 contains six putative Protein Kinase C serine/threonine phosphorylation sites, we examined the ability of PKC to phosphorylate Ebp1 and to regulate Ebp1-ErbB3 binding. We found that Ebp1 was basally phosphorylated in AU565 breast cancer cells on serine/threonine residues and that this phosphorylation was enhanced by heregulin treatment. Both serine and threonine residues of a GST-Ebp1 fusion protein were phosphorylated by PKC in vitro. In vivo, we demonstrated that basal Ebp1 phosphorylation was dependent upon PKC. However, HRG-induced phosphorylation of Ebp1 occurred predominantly in a PKC-independent manner. The ability of Ebp1 to associate with ErbB3 in serum-starved NIH3T3 cells overexpresssing ErbB3 was abrogated by treating cells with a PKC inhibitor. These findings suggest that PKC plays a role in regulating phosphorylation and function of Ebp1 in vivo.
Lessor
T J
TJ
Molecular and Cellular Biology Program, University of Maryland, Baltimore, MD 21201, USA.
Hamburger
A W
AW
D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002352:Carrier Proteins / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D051379:Mice; D020890:Neuregulin-1 / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism / Q000494:pharmacology*; D016601:RNA-Binding Proteins; D020893:Receptor, ErbB-3 / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D020890:Neuregulin-1; C101633:PA2G4 protein, human; D016601:RNA-Binding Proteins; D020893:Receptor, ErbB-3; D011493:Protein Kinase C