Sequence-specific interaction of a conformational domain of p53 with DNA.
Mutations within a conserved "conformational" domain of the p53 protein 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-binding motif. Hybrid proteins containing mutations in p53 bound to DNA less efficiently than wild-type hybrid protein. In addition, competition experiments showed that mutated p53 DNA-binding motif failed to interact with p53 hybrid proteins. The DNA-binding activity of wild-type p53 hybrid protein was inhibited by the metal chelator 1,10-phenanthroline. These results demonstrate that DNA-binding activity resides in the conformational domain of p53, providing a structural model for disruption of DNA binding by mutation. Furthermore, metal ions may regulate binding of p53 to DNA by modulating its conformation.
Srinivasan
R
R
Department of Thoracic and Cardiovascular Surgery, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Roth
J A
JA
Maxwell
S A
SA
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D002417:Cattle; D017124:Conserved Sequence; D004247:DNA / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D013539:Simian virus 40; D013205:Staphylococcal Protein A / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000952:Antigens, Polyomavirus Transforming; D013205:Staphylococcal Protein A; D016159:Tumor Suppressor Protein p53; D004247:DNA