RPG: Towards the Crystal Structure of Replication Protein A

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9708028 Borgstahl This is a Research Planning Grant for Women Scientists and Engineers. Dr. Borgstahl will study the structure of replication protein A (RPA) by X-ray crystallography. RPA is essential to the maintenance and replication of DNA. The recombinant protein will be produced in E.coli and very pure protein samples will be obtained using the latest techniques and established protocols. A wide range of crystallization conditions will be explored to grow large well-ordered crystals. Three strategies will be used to obtain diffraction quality crystals: crystallization of intact yeast and human RPA; crystallization of RPA domains and subcomplexes; crystallization of a trimmed RPA heterotrimer. Once crystals are obtained, X-ray diffraction data will be collected in order to obtain a high resolution structure of the protein. Replication protein A is a fundamental molecule in metabolism and is essential for eukaryotic life. RPA is absolutely required for many DNA metabolic processes, including replication, nucleotide excision repair and homologous recombination. The elucidation of the three-dimensional structure of RPA should provide insight into how this protein interacts with DNA and how it functions in its essential multidisciplinary role in DNA metabolism. ***