Rational design of an estrogen receptor mutant with altered DNA-binding specificity
Open Access
- 22 April 2007
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 35 (10), 3465-3477
- https://doi.org/10.1093/nar/gkm241
Abstract
Although artificial C2-H2 zinc fingers can be designed to recognize specific DNA sequences, it remains unclear to which extent nuclear receptor C4 zinc fingers can be tailored to bind novel DNA elements. Steroid receptors bind as dimers to palindromic response elements differing in the two central base pairs of repeated motifs. Predictions based on one amino acid—one base-pair relationships may not apply to estrogen receptors (ERs), which recognize the two central base pairs of estrogen response elements (EREs) via two charged amino acids, each contacting two bases on opposite DNA strands. Mutagenesis of these residues, E203 and K210 in ERα, indicated that both contribute to ERE binding. Removal of the electric charge and steric constraints associated with K210 was required for full loss of parental DNA-binding specificity and recognition of novel sequences by E203 mutants. Although some of the new binding profiles did not match predictions, the double mutation E203R-K210A generated as predicted a mutant ER that was transcriptionally active on palindromes of PuGCTCA motifs, but not on consensus EREs. This study demonstrates the feasibility of designing C4 zinc finger mutants with novel DNA-binding specificity, but also uncovers limitations of this approach.Keywords
This publication has 39 references indexed in Scilit:
- Structural determinants of nuclear receptor assembly on DNA direct repeatsNature, 1995
- DNA recognition code of transcription factors in the helix-turn-helix, probe helix, hormone receptor, and zinc finger families.Proceedings of the National Academy of Sciences of the United States of America, 1994
- Toward a code for the interactions of zinc fingers with DNA: selection of randomized fingers displayed on phage.Proceedings of the National Academy of Sciences of the United States of America, 1994
- Differential Recognition of Target Genes by Nuclear Receptor Monomers, Dimers, and Heterodimers*Endocrine Reviews, 1994
- Refined solution structure of the glucocorticoid receptor DNA-binding domainBiochemistry, 1993
- The crystal structure of the estrogen receptor DNA-binding domain bound to DNA: How receptors discriminate between their response elementsCell, 1993
- Defining a minimal estrogen receptor DNA binding domainNucleic Acids Research, 1993
- Multiple parameters control the selectivity of nuclear receptors for their response elements. Selectivity and promiscuity in response element recognition by retinoic acid receptors and retinoid X receptors.Journal of Biological Chemistry, 1993
- Structural Determinants of a Glucocorticoid Receptor Recognition ElementMolecular Endocrinology, 1990
- The Uteroglobin Promoter Contains a Noncanonical Estrogen Responsive ElementMolecular Endocrinology, 1990