The goal of this project is an atomic-level description of the mechanism by which a yeast zinc finger transcription factor, ADR1, controls transcription of the ADH2 gene in Saccharomyces cerevisae. Structural, dynamic, and energetic aspects of two processes central to transcriptional activation will be investigated using high resolution NMR spectroscopy, complemented by thermodynamic measurements using microcalorimetry: (1) sequence-specific binding by the DNA binding domain of ADR1 to its cognate DNA, the upstream activator sequence, UAS1, and (2) structural properties of an identified activation domain in ADR1. The initial aims include a complete characterization of the minimal DNA-binding domain of ADR1, in both its free form and when bound to a UAS1 DNA site. The domain structure of the protein will be characterized by limited proteolysis and MALDI-mass spectrometry.
| Schaufler, Lawrence E; Klevit, Rachel E (2003) Mechanism of DNA binding by the ADR1 zinc finger transcription factor as determined by SPR. J Mol Biol 329:931-9 |
| Hyre, D E; Klevit, R E (1998) A disorder-to-order transition coupled to DNA binding in the essential zinc-finger DNA-binding domain of yeast ADR1. J Mol Biol 279:929-43 |
| Schmiedeskamp, M; Rajagopal, P; Klevit, R E (1997) NMR chemical shift perturbation mapping of DNA binding by a zinc-finger domain from the yeast transcription factor ADR1. Protein Sci 6:1835-48 |
| Schmiedeskamp, M; Klevit, R E (1997) Paramagnetic cobalt as a probe of the orientation of an accessory DNA-binding region of the yeast ADR1 zinc-finger protein. Biochemistry 36:14003-11 |
