SEQUENCE-SPECIFIC INTERACTION OF A CONFORMATIONAL DOMAIN OF P53 WITH DNA

Mutations within a conserved ''conformational'' domain of the p53 prot ein have frequently been observed in a wide variety of human cancers. A hybrid protein containing the wild-type conformational domain of p53 fused to protein A bound to calf thymus DNA and a specific p53 DNA-bi nding motif. Hybrid proteins containing mutations in p53 bound to DNA less efficiently than wild-type hybrid protein. In addition, competiti on experiments showed that mutated p53 DNA-binding motif failed to int eract with p53 hybrid proteins. The DNA-binding activity of wild-type p53 hybrid protein was inhibited by the metal chelator 1,10-phenanthro line. These results demonstrate that DNA-binding activity resides in t he conformational domain of p53, providing a structural model for disr uption of DNA binding by mutation. Furthermore, metal ions may regulat e binding of p53 to DNA by modulating its conformation.