NMR chemical shift perturbation mapping of dna binding by a zinc‐finger domain from the yeast transcription factor ADR1

• DOI: 10.1002/pro.5560060904
• Publication Date: 2009-02-10T06:07:37Z
• Authors: Mia Schmiedeskamp; Ponni Rajagopal; Rachel E. Klevit
• Keywords: RING finger domain; Zinc finger nuclease; LIM domain; Chemical shift; HMG-box; Protein–DNA interaction

Abstract Mutagenesis studies have revealed that the minimal DNA‐binding domain of yeast transcription factor ADR1 consists two Cys 2 ‐His zinc fingers plus an additional 20 residues proximal and N‐terminal to fingers. We assigned NMR 1 'H, 15 N, 13 C chemical shifts for entire both free bound specific DNA. H shift values suggest little structural difference between in this construct single‐finger constructs, α index analysis indicates change finger structure upon DNA binding. perturbations binding are observed, however, these mapped define protein‐DNA interface. The appear bind with different orientations, as helix is perturbed, while only extreme N‐terminus affected. Furthermore, first undergo large changes suggesting a role at A striking correspondence observed interface by previously mutagenesis.