Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90.

• DOI: 10.1073/pnas.93.16.8379
• Publication Date: 2002-07-26T14:34:16Z
• Authors: M V Blagosklonny; J Toretsky; S Bohen; L Neckers
• Keywords: Geldanamycin; Reticulocyte; Wild type; Hsp90 inhibitor; Chaperone (clinical)

The p53 mutant, 143ala, was translated in vitro either rabbit reticulocyte lysate (RRL) or wheat germ extract (WGE). In RRL, p53-143ala protein of both mutant and wild-type conformation, as detected immunologically with conformation-specific antibodies, translated. chaperone HSP90, present found to coprecipitate only the mutated conformation p53. Geldanamycin, shown previously bind HSP90 destabilize its association other proteins, decreased amount detectable increased protein, without affecting total translation When WGE, known contain functionally deficient produced which not recognized by a antibody. contrast, synthesis conformationally this system compromised. Reconstitution function WGE permitted p53, abrogated geldanamycin. Finally, when stably tansfected into yeast engineered be defective for function, conformational recognition impaired. stable transfectants were prepared expressing antagonized macbecin I, geldanamycin analog also HSP90. Taken together, these data demonstrate role achievement and/or stabilization p53-143ala. Furthermore, we show that can pharmacologically drugs targeting