title: E5 oncoprotein retained in the endoplasmic reticulum/cis Golgi still induces PDGF receptor autophosphorylation but does not transform cells.
abstract: The E5 oncoprotein encoded by bovine papillomavirus type 1 is a homodimeric, hydrophobic polypeptide which is localized predominantly in Golgi membranes and which transforms several cell types apparently by inducing tyrosine phosphorylation of the platelet-derived growth factor receptor (PDGF-R). While the precise mechanism of receptor activation is unknown, E5 associates with several cellular proteins, including PDGF-R and the 16K V-ATPase protein, and induces the preferential phosphorylation of immature, Endo H-sensitive forms of the receptor. To evaluate whether E5 accumulation in the Golgi was requisite for receptor phosphorylation and cell transformation, we sequestered the E5 protein in the endoplasmic reticulum (ER)/cis Golgi by appending the ER retention KDEL sequence to its C-terminus. In transient assays and in cell lines, E5/KDEL protein and E5/KDEL* protein (a defective variant of KDEL), were stable and formed homodimers normally. E5/KDEL*, similar to wt E5, localized to the Golgi and was transformation-proficient. In contrast, E5/KDEL failed to concentrate in the Golgi and was transformation-incompetent. Despite these critical defects, however, E5/KDEL formed stable complexes with immature PDGF-R and 16K and, even more unexpectedly, induced the phosphorylation of both mature and immature PDGF-R on tyrosine residues to the same level as wt E5. These data demonstrate that E5 can bind and induce PDGF-R phosphorylation in the ER/cis Golgi, but that successful mitogenic signalling (and consequent cell transformation) requires the translocation of E5/receptor complexes to distal Golgi compartments.
authors.affiliation: Department of Pathology, Georgetown University, Washington, DC 20007, USA.
authors.forename: J
authors.initials: J
authors.lastname: Sparkowski
authors.forename: J
authors.initials: J
authors.lastname: Anders
authors.forename: R
authors.initials: R
authors.lastname: Schlegel
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009842:Oligopeptides / Q000096:biosynthesis / Q000235:genetics; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D021382:Protein Sorting Signals; D006180:Proton-Translocating ATPases / Q000378:metabolism; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
chemical_list: D009842:Oligopeptides; D009856:Oncogene Proteins, Viral; D021382:Protein Sorting Signals; C054304:oncogene protein E5, Bovine papillomavirus type 1; C073762:lysyl-aspartyl-glutamyl-leucine; D014443:Tyrosine; D017479:Receptors, Platelet-Derived Growth Factor; D006180:Proton-Translocating ATPases