DNA binding specificity of mutant glucocorticoid receptor DNA-binding domains.

摘要: Mutation of a small number amino acids in the DNA-binding domain estrogen receptor to corresponding sequence glucocorticoid switches specificity transactivation assays (Mader, S., Kumar, V., de Verneuil, H., and Chambon, P. (1989) Nature 338, 271-274). We have made reciprocal mutations context studied binding wild type mutant purified proteins palindromic response elements as well intermediate sequence, using gel mobility shift assays. show here that protein with two altered binds low but equal affinity, whereas an additional changed residue has high affinity for still retains considerable elements. Using sites we further characterized interaction DNA. The vitro DNA results are confirmed by vivo yeast. Finally suggest testable model acid/base pair interactions involved recognition its target sequence.