Induction of gene amplification as a gain-of-function phenotype of mutant p53 proteins.
Gene amplification accompanies tumor progression and is involved in the development of drug resistance. Previously, we reported (A. Albor et al., Cancer Res. 58: 2091-2094, 1998) that mutant p53 proteins conserve the capacity to interact with and activate topoisomerase I, and that this could be a mechanism for induction of genomic instability by mutant p53 proteins. To test this hypothesis, the effect of exogenous mutant p53 protein expression on genomic instability in human p53-/- Saos-2 cells was measured by the frequency of formation of N-(phosphoacetyl)-L-aspartate (PALA)-resistant (PALA(R)) colonies, mediated by gene amplification. Interaction of exogenous mutant p53 and topoisomerase I was confirmed by immunoprecipitation. Growth under continuous expression of mutant p53 proteins for 16-17 population doublings increased the frequency of appearance of PALA(R) colonies after subsequent exposure to PALA. Subtoxic concentrations of camptothecin (which stabilizes topoisomerase I cleavage complexes, mediating nonhomologous recombination) produced a dose-dependent increase in PALA(R) colonies, and combining expression of mutant p53 with exposure to camptothecin produced a greater than additive increase in PALA(R) colony formation. These results indicate that mutant p53 proteins promote gene amplification independently of their capacity to inactivate the wild-type p53 protein, and suggest that this effect is dependent on interaction of mutant p53 with topoisomerase I. Additional studies are needed to assess the potential of targeting mutant p53 interaction with topoisomerase I for the reduction of drug resistance development during chemotherapy.
El-Hizawi
Sally
S
Department of Dermatology, Oregon Health and Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97201, USA.
Lagowski
James P
JP
Kulesz-Martin
Molly
M
Albor
Amador
A
D001224:Aspartic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D001859:Bone Neoplasms / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D002166:Camptothecin / Q000494:pharmacology; D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005784:Gene Amplification / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D009154:Mutation; D012516:Osteosarcoma / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010746:Phosphonoacetic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D059004:Topoisomerase I Inhibitors; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D004791:Enzyme Inhibitors; D059004:Topoisomerase I Inhibitors; D016159:Tumor Suppressor Protein p53; D001224:Aspartic Acid; C013195:sparfosic acid; D004264:DNA Topoisomerases, Type I; D010746:Phosphonoacetic Acid; D002166:Camptothecin