Core 2: Structural Cell Biology (SCB) Core

摘要-核心2：结构细胞生物学(SCB) SBDR4中癌症结构生物学的一个主要挑战是从结构上定义DNA修复(DR)蛋白 以及维持基因组完整性的组件。然而。DR蛋白和复合体具有编码灵活性 并在空间和时间协调的复合体中发挥作用，这使得它们的结构特征如此 很有挑战性。凭借SBDR调查人员，结构细胞生物学(SCB)核心克服了这些问题 通过提供对尖端结构生物学技术的获取和专业知识来应对极端挑战 需要探测DR蛋白质和复合体，个别研究机构没有提供。 目的1.确定结构、构象和组装状态。渣打银行核心，以国家 同步加速器设施，将提供大分子结晶学(MX)和小角X射线散射(SAXS) 为SBDR成员提供数据收集和光束线科学家级别的专业知识。MX提供高分辨率原子 SAXS可以快速评估灵活性、溶液构象(S)和组件。SCB SAXS 包括通过GOLD-SAXS对配体或缓冲液进行筛选和评估DNA弯曲。 目的2.检测整合结构生物学中的褶皱和构象。大连湾大型综合体的分析 由兆道顿结构质谱仪(MS)和SAXS验证的结构预测的项目将 他们自己提供可测试的假说，或者将加速个别研究所的低温电子显微镜研究。 目的3.通过同步加速器软X射线确定重组系统和细胞中的分子结构 重组相变和细胞的层析成像(SXT)和超分辨荧光 DR组件的定量测量，否则将是有问题的 传统的结构方法。 SCB核心位于高级光源(ALS)同步加速器的中心，其中三条光束线(12.3.1， 8.3.1和国家X射线层析成像中心/NCXT)将提供SAXS、MX和SXT数据收集 SBDR成员。为了实现对其他结构技术和功能的访问，SCB设有分支机构 在卡尔加里(MS)、纽约(超分辨率显微镜)、牛津(MX)和斯克里普斯(MX)。 所有四个项目都将使用SCB核心，无论成员实验室的结构专业知识如何，以最佳方式 并以典型的实验室和光束线有效地满足项目的结构和机械目标 访问不能也不能做。总而言之，这些SCB模式允许SBDR-5询问动态DR 从原子到细胞级别的机器。SCB核心的广泛、长期目标是支持SBDR 确定序列变化的影响的项目，剖析多种功能的方法，以及 机制，从而灵活和动态的DR机械功能。

SUMMARY - CORE 2: STRUCTURAL CELL BIOLOGY (SCB) A major challenge for cancer structural biology in SBDR4 is to structurally define the DNA repair (DR) proteins and assemblies that maintain genomic integrity. However. DR proteins and complexes have encoded flexibility and function in spatially and temporally coordinated complexes that make their structural characterizations so challenging. With SBDR investigators, Core 2, the Structural Cell Biology (SCB) Core overcomes these extreme challenges by providing access to and expertise on cutting-edge structural biology technologies needed to probe DR proteins and complexes and not available in individual research institutions. Aim 1. Determine structures, conformations and assembly states. The SCB Core, centered at a national synchrotron facility, will provide macromolecular crystallography (MX) and small angle X-ray scattering (SAXS) data collection and beamline scientist-level expertise for SBDR members. MX provides high resolution atomic structures, while SAXS can rapidly assess flexibility, solution conformation(s), and assemblies. SCB SAXS includes ligand or buffer screening and assessing DNA bending, via GOLD-SAXS. Aim 2. Detect folds and conformations for integrative structural biology. Analysis of large complexes in the Projects, by megadalton structural mass spectrometry (MS) and with SAXS-validated structure predictions, will provide testable hypotheses on their own, or will accelerate CryoEM studies at individual institutes. Aim 3. Define molecular architectures in reconstituted systems and in the cell through synchrotron soft X-ray tomography (SXT) of reconstituted phase transitions and cells and super-resolution fluorescence measurements for quantitative measurements of DR assemblies, that would otherwise be problematic for traditional structural methods. The SCB Core is centered at the Advanced Light Source (ALS) synchrotron, where three beamlines (12.3.1, 8.3.1, and National Center for X-ray Tomography/NCXT) will provide SAXS, MX, and SXT data collection to SBDR members. For enabling access to additional structural technologies and capabilities, SCB has branches at Calgary (MS), New York (super-resolution microscopy), Oxford (MX), and Scripps (MX). All four Projects will use the SCB Core, regardless of structural expertise in member laboratories, to optimally and efficiently address Project structural and mechanistic Aims in ways that typical laboratories and beamline visits cannot or do not do. Together, these SCB modalities allow SBDR-5 interrogation of dynamic DR machines from atomic to cellular scales. The broad, long range objective of the SCB Core is to support SBDR Projects to determine the impact of sequence variations, the means to dissect multiple functions, and the mechanisms whereby flexible and dynamic DR machinery functions.