Ethylation interference and X-ray crystallography identify similar interactions between 434 repressor and operator.
In the crystal structure of a repressor-operator complex (the 434 repressor DNA-binding domain and its 14-base pair (bp) operator), Anderson et al. elsewhere in this issue identify six positions of likely contact between repressor protein and phosphates of the DNA backbone. At each of these positions, electron densities of protein and DNA merge. Experiments presented here indicate that intact 434 repressor approaches these phosphates very closely when it is bound to DNA in solution. Specifically, when any one of these phosphates is ethylated, repressor cannot bind to the modified operator. We also identify another position where ethylation has a significant but less dramatic effect on repressor binding, and note that in the structure, repressor closely approaches this phosphate. Our results strongly support the idea that the interactions between protein and the DNA phosphate backbone in the crystallized complex are the same as those made by intact repressor to operator DNA in solution. In addition, our results suggest that DNA is slightly bent by repressor binding.
D000478:Alkylation; D004247:DNA; D009690:Nucleic Acid Conformation; D009876:Operon; D012097:Repressor Proteins; D014157:Transcription Factors; D014961:X-Ray Diffraction
D012097:Repressor Proteins; D014157:Transcription Factors; D004247:DNA