Selective binding of different p53 response elements by p53 containing complexes.

Abstract

The p53 tumor suppressor protein binds two copies of a ten base pair motif that is degenerate in eight out of ten bases and conforms to the sequence, 5'PuPuPuC(A/T)(T/A)GPyPyPy-3'. As a consequence of this high degree of degeneracy, p53 response elements show a great deal of variation and it has been speculated that the variation aids in the selective activation of p53 responsive genes by specific stimuli. Here, we examined the DNA binding characteristics of several different p53 protein complexes present in nuclear extracts prepared from a cell line expressing the murine temperature sensitive p53 protein, p53val135. Interestingly, the complexes exhibited a distinct preference for binding to some p53 response elements and not others. A critical determinant of this specificity was the sequence at the center of the ten base pair motif and alteration of a single base within this region was sufficient to alter the set of complexes that associated with the oligonucleotide. In addition, thermal denaturation experiments demonstrated that some complexes could bind DNA even though the p53val135 protein had a mutant conformation. Our results are consistent with the hypothesis that p53 can distinguish between various response elements and suggest that this selectivity is manifested, in part, by the sequence of the motif and conformation of the p53 protein.