title: Temperature-sensitive mutants of p53 associated with human carcinoma of the lung.
abstract: We have compared the effects of specific point mutations on the tertiary and quaternary structure of the human p53 protein. Eight mutants, each derived from primary resected tissues of lung carcinomas, were expressed in vitro under strictly defined conditions, such that the only known variant was the point mutation present in each p53 mRNA. All the mutations were located in highly conserved domains. The tertiary structure of each mutant protein was investigated by reactivity with anti-p53 monoclonal antibodies directed against conformation-dependent epitopes. Quaternary structure was examined by gel filtration. Although all the mutant proteins exhibited abnormal tertiary structures, their quaternary structures appeared similar to wild type, the one exception being p53-tyr135, which contains tyrosine in place of cysteine at residue 135. The conformational phenotype of mutant human p53 was found to be dependent upon (i) the locus of the mutation and (ii) the nature of the amino acid substitution: two different substitutions at residue 273 yielded two mutants with differing structural properties. We have discovered three mutants of human p53 that are temperature sensitive for conformation; one is mutated at codon 273, a 'hotspot' for p53 mutation in human cancer.
authors.affiliation: Department of Pathology, University of Cambridge, UK.
authors.forename: E A
authors.initials: EA
authors.lastname: Medcalf
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
authors.forename: I
authors.initials: I
authors.lastname: Chiba
authors.forename: J
authors.initials: J
authors.lastname: Minna
authors.forename: J
authors.initials: J
authors.lastname: Milner
mesh_terms: D002277:Carcinoma / Q000737:chemistry*; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000737:chemistry*; D009154:Mutation / Q000235:genetics*; D010641:Phenotype; D014176:Protein Biosynthesis; D011487:Protein Conformation; D013696:Temperature; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000737:chemistry*
chemical_list: D016159:Tumor Suppressor Protein p53