Tumor suppressor p53: analysis of wild-type and mutant p53 complexes.
It has been suggested that the dominant effect of mutant p53 on tumor progression may reflect the mutant protein binding to wild-type p53, with inactivation of suppressor function. To date, evidence for wild-type/mutant p53 complexes involves p53 from different species. To investigate wild-type/mutant p53 complexes in relation to natural tumor progression, we sought to identify intraspecific complexes, using murine p53. The mutant phenotype p53-246(0) was used because this phenotype is immunologically distinct from wild-type p53-246+ and thus permits immunological analysis for wild-type/mutant p53 complexes. The p53 proteins were derived from genetically defined p53 cDNAs expressed in vitro and also from phenotypic variants of p53 expressed in vivo. We found that the mutant p53 phenotype was able to form a complex with the wild type when the two p53 variants were cotranslated. When mixed in their native states (after translation), the wild-type and mutant p53 proteins did not exhibit any binding affinity for each other in vitro. Under identical conditions, complexes of wild-type human and murine p53 proteins were formed. For murine p53, both the wild-type and mutant p53 proteins formed high-molecular-weight complexes when translated in vitro. This oligomerization appeared to involve the carboxyl terminus, since truncated p53 (amino acids 1 to 343) did not form complexes. We suggest that the ability of the mutant p53 phenotype to complex with wild type during cotranslation may contribute to the transforming function of activated mutants of p53 in vivo.
Milner
J
J
Department of Pathology, University of Cambridge, United Kingdom.
Medcalf
E A
EA
Cook
A C
AC
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D003001:Cloning, Molecular; D016147:Genes, Tumor Suppressor; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D009154:Mutation; D011233:Precipitin Tests; D011485:Protein Binding; D013045:Species Specificity; D013329:Structure-Activity Relationship; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000276:immunology
D000911:Antibodies, Monoclonal; D046911:Macromolecular Substances; D016159:Tumor Suppressor Protein p53