Biochemical characterization of a nuclear factor that binds to NF1-like elements in the rat p53 promoter.
We previously reported that two nuclear factor 1-like elements mediated the transcription of the rat p53 gene. A 40-kDa protein was shown to bind to these elements, which was different from common NF1 family proteins. In this study, the biochemical properties of the 40-kDa binding protein were investigated. The metal ion dependency of the protein was examined with various chelators; the protein was proved to require Mg(2+) for maximum DNA-binding activity. The binding protein was highly resistant to ionic strength and denaturant. The protein-DNA complex was reduced at high NaCl concentration, but residual DNA-binding activity remained. Even 2 M urea did not completely eliminate the formation of protein-DNA complex. DNA-binding activity of the protein was also stable at high temperature. Treatment of the protein-DNA complex with increasing concentrations of proteinase K or trypsin demonstrated the existence of a protease-resistant DNA-bound core. These biochemical properties provide new insight into the 40-kDa NF1-like nuclear factor.