Supporting data for “<i>Structural and Functional Characterization of the Eukaryotic Replisome</i>”

The dataset provided is associated with the doctoral thesis titled "<i>Structural and Functional Characterization of the Eukaryotic Replisome</i>". 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 <i>in vitro</i> 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.

本数据集关联于题为《真核复制体(Eukaryotic Replisome)的结构与功能表征》的博士学位论文。本综合性数据集涵盖了用于探究真核复制体的各类实验技术与研究方法。该项研究主要致力于解析CMG解旋酶(CMG Helicase)转位过程中其与聚合酶ε(Polymerase ε)之间的协同互作关系。本数据集覆盖了该项研究的多维度内容，涵盖蛋白质纯化、生化重构、结构解析以及功能检测四大板块。研究团队采用蛋白质纯化技术，以分离获取高纯度的复制体组分样本。该纯化流程包含多步操作：如过表达、亲和纯化以及色谱层析，用以从复杂生物样本中分离并富集复制体相关蛋白。生化重构是该项研究的另一关键环节：研究人员将纯化得到的复制体组分在可控的体外(in vitro)环境中进行组装，以重现其功能性互作与活性状态。通过重构复制体，本研究旨在解析其三维结构并阐明调控DNA复制的核心分子机制。本数据集同时涵盖结构解析类实验：此类实验借助冷冻电镜(Cryo-EM)技术，以解析复制体的三维空间结构。功能检测实验则用于评估复制体的酶促活性与功能特性，包括DNA解旋、DNA合成以及CMG解旋酶与聚合酶ε之间的互作关系。综上，本数据集是在真核复制体研究过程中产生的实验数据的综合性集合，涵盖蛋白质纯化、生化重构、结构解析与功能检测等全部环节，可为解析DNA复制的分子机制提供极具价值的研究资源。

搜集汇总

背景与挑战

背景概述

该数据集是博士论文《Structural and Functional Characterization of the Eukaryotic Replisome》的配套数据，发布于2023年9月19日，作者为Zhichun Xu。数据集涵盖了蛋白质纯化、生化重构、结构测定和功能分析等多种实验技术，重点研究真核复制体中CMG解旋酶与聚合酶ε在解旋酶转运过程中的协调机制，属于结构生物学领域，并采用CC BY-NC 4.0许可证共享。