Genomics research is providing unprecedented access to expression vectors for thousands of proteins and it is critical to develop faster structure-determination technology to discover their cellular roles. Solution-state nuclear magnetic resonance (NMR) spectroscopy is a powerful and widely used method to determine accurate protein structures, but is also time consuming. Progress in understanding eukaryotic mechanisms of replication, transcription and translation depends critically on gaining faster access to structures of domains and proteins identified in human and other genome projects. We propose new technology for fast protein structure determination by solution state NMR. We will develop and demonstrate this technology on the 127 residue bromodomain from yeast Sth1p. Bromodomain has been chosen for its relevance to the regulation of acetylation states of histones in eukaryotic DNA. The BrD stucture will yield insights into its roles in transcription activation and repression protein complexes and should help to clarify the role of acetylation regulation in carcinogenesis. We will transfer this technology to demonstrate high throughput determination of multiple structures associated with breast cancer gene sequences in collaboration with the newly established Harvard Structural Genomics of Cancer Initiative. We expect that discovery of new eukaryotic protein structures will lead to a reevaluation of targets identified for drug design and to new directions for cancer studies.
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