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Supporting data for “Structural and Functional Characterization of the Eukaryotic Replisome”
The dataset provided is associated with the doctoral thesis titled "Structural and Functional Characterization of the Eukaryotic Replisome". This comprehensive dataset encompasses various experimental techniques and methodologies employed to investigate the eukaryotic replisome. The research primarily focuses on understanding the coordination between the CMG helicase and polymerase epsilon during the helicase translocation.
The dataset encompasses diverse aspects of the research, including protein purification, biochemical reconstitution, structure determination, and functional assays. Protein purification techniques were employed to isolate and obtain highly pure samples of the replisome components. This process involved various purification steps, such as over-expression, affinity based purification and chromatography, to separate and enrich the replisome proteins from complex biological samples. Biochemical reconstitution was another crucial aspect of the research. It involved the assembly of the purified replisome components in a controlled in vitro environment to recapitulate their functional interactions and activities. By reconstituting the replisome, we aimed to solve the structure and understand the underlying molecular mechanisms governing DNA replication. The dataset also includes structure determination experiments. These experiments utilized techniques cryo-EM to determine the three-dimensional structures of the replisome. Functional assays evaluated the enzymatic activities and functional properties of the replisome, such as DNA unwinding, DNA synthesis, and interaction between CMG and polymerase epsilon. Overall, this dataset represents a comprehensive collection of experimental data generated during the investigation of the eukaryotic replisome. It encompasses protein purification, biochemical reconstitution, structure determination, and functional assays, providing a valuable resource for understanding the mechanism.
