title: The physical association of multiple molecular chaperone proteins with mutant p53 is altered by geldanamycin, an hsp90-binding agent.
abstract: Wild-type p53 is a short-lived protein which turns over very rapidly via selective proteolysis in the ubiquitin-proteasome pathway. Most p53 mutations, however, encode for protein products which display markedly increased intracellular levels and are associated with positive tumor-promoting activity. The mechanism by which mutation leads to impairment of ubiquitination and proteasome-mediated degradation is unknown, but it has been noted that many transforming p53 mutants are found in stable physical association with molecular chaperones of the hsp70 class. To explore a possible role for aberrant chaperone interactions in mediating the altered function of mutant p53 and its intracellular accumulation, we examined the chaperone proteins which physically associate with a temperature-sensitive murine p53 mutant. In lysate prepared from A1-5 cells grown under mutant temperature conditions, hsp70 coprecipitated with p53Val135 as previously reported by others, but in addition, other well-recognized elements of the cellular chaperone machinery, including hsp90, cyclophilin 40, and p23, were detected. Under temperature conditions favoring wild-type p53 conformation, the coprecipitation of chaperone proteins with p53 was lost in conjunction with the restoration of its transcriptional activating activity. Chaperone interactions similar to those demonstrated in A1-5 cells under mutant conditions were also detected in human breast cancer cells expressing two different hot-spot mutations. To examine the effect of directly disrupting chaperone interactions with mutant p53, we made use of geldanamycin (GA), a selective hsp90-binding agent which has been shown to alter the chaperone associations regulating the function of unliganded steroid receptors. GA treatment of cells altered heteroprotein complex formation with several different mutant p53 species. It increased p53 turnover and resulted in nuclear translocation of the protein in A1-5 cells. GA did not, however, appear to restore wild-type transcriptional activating activity to mutant p53 proteins in either A1-5 cells or human breast cancer cell lines.
authors.affiliation: Department of Pediatrics and Steele Memorial Children's Research Center, University of Arizona, Tucson 85724, USA. whitelj@peds.arizona.edu
authors.forename: L
authors.initials: L
authors.lastname: Whitesell
authors.forename: P D
authors.initials: PD
authors.lastname: Sutphin
authors.forename: E J
authors.initials: EJ
authors.lastname: Pulcini
authors.forename: J D
authors.initials: JD
authors.lastname: Martinez
authors.forename: P H
authors.initials: PH
authors.lastname: Cook
mesh_terms: D000818:Animals; D016227:Benzoquinones; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins; D006801:Humans; D019746:Intramolecular Oxidoreductases; D047029:Lactams, Macrocyclic; D051379:Mice; D018832:Molecular Chaperones / Q000378:metabolism; D009154:Mutation; D019696:Peptidylprolyl Isomerase / Q000187:drug effects / Q000378:metabolism*; D010750:Phosphoproteins / Q000187:drug effects / Q000378:metabolism*; D000071876:Prostaglandin-E Synthases; D011809:Quinones / Q000494:pharmacology*; D051381:Rats; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism
chemical_list: D016227:Benzoquinones; D002352:Carrier Proteins; D000081406:Peptidyl-Prolyl Isomerase F; D004791:Enzyme Inhibitors; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D018832:Molecular Chaperones; C000631446:PPIF protein, mouse; D010750:Phosphoproteins; D011809:Quinones; D016159:Tumor Suppressor Protein p53; D021983:Cyclophilins; D019696:Peptidylprolyl Isomerase; D019746:Intramolecular Oxidoreductases; C490849:PTGES3 protein, human; D000071876:Prostaglandin-E Synthases; C001277:geldanamycin