title: pRb, Myc and p53 are critically involved in SV40 large T antigen repression of PDGF beta-receptor transcription.
abstract: The expression of the PDGF beta-receptor is tightly regulated during a normal cell cycle. c-Myc and p73alpha repress transcription of the receptor through interaction with NF-Y. In ST15A cells which stably express the temperature-sensitive SV40 large T antigen (LT) the receptor expression and ligand binding decreased under the permissive condition. Transient expression of the LT, but not small t, decreased the endogenous receptor expression at mRNA and protein levels in NIH3T3 cells but not in the myc-null HO15.19 cells. The wild-type LT, but not the various pRb or p53 binding defective LT mutants, represses the PDGF beta-receptor promoter activity. Moreover, the inability of the LT-mediated repression in the myc-null cells, the Rb-null 3T3 cells, and the Saos-2 cells lacking pRb and p53, indicates that Myc, pRb and p53 are all necessary elements. PDGF beta-receptor promoter-luciferase assays revealed that the CCAAT motif is important for the repression. Furthermore, p53 was found to increase the promoter activity mainly via the upstream Sp1 binding sites together with the CCAAT motif in the NIH 3T3 cells. This was confirmed by Schneider's Drosophila line (SL2) cells deficient in both endogenous NF-Y and Sp1. Chromatin immunoprecipitation using ST15A cells revealed that both LT and p53 bound the PDGF beta-receptor promoter and the binding of p53 diminished when LT was expressed in the permissive condition. However, LT binds the promoter in the absence of pRb and p53 in Saos-2 cells stably expressing LT. These results suggest that LT binds the promoter and interferes with NF-Y and Sp1 to repress it in the presence of Myc, pRb and p53.
authors.affiliation: Department of Cell Biology, Institute of Anatomy and Cell Biology, Göteborg University, Box 420, SE-405 30 Gothenburg, Sweden.
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Uramoto
authors.forename: Anders
authors.initials: A
authors.lastname: Hackzell
authors.forename: Daniel
authors.initials: D
authors.lastname: Wetterskog
authors.forename: Andrea
authors.initials: A
authors.lastname: Ballági
authors.forename: Hiroto
authors.initials: H
authors.lastname: Izumi
authors.forename: Keiko
authors.initials: K
authors.lastname: Funa
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry* / Q000378:metabolism; D023081:CCAAT-Binding Factor / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D015536:Down-Regulation; D004330:Drosophila; D017930:Genes, Reporter; D015151:Immunoblotting; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D009154:Mutation; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism*; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism*; D016160:Retinoblastoma Protein / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016329:Sp1 Transcription Factor / Q000378:metabolism; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D000952:Antigens, Polyomavirus Transforming; D023081:CCAAT-Binding Factor; D008024:Ligands; D016271:Proto-Oncogene Proteins c-myc; D016160:Retinoblastoma Protein; D016329:Sp1 Transcription Factor; D016159:Tumor Suppressor Protein p53; D008156:Luciferases; D020797:Receptor, Platelet-Derived Growth Factor beta