Core 2: Structural Cell Biology (SCB) Core

核心 2：结构细胞生物学 (SCB) 核心

• 批准号: 10271098
• 负责人: Gregory L Hura
• 金额: $ 74.51万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-27 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271098
• 关键词: Active Sites; Address; Architecture; Binding; Buffers; Cells; Cellular Structures; Cellular biology; Complement; Complex; Coupled; Cryoelectron Microscopy; Crystallization; Crystallography; DNA; DNA Probes; DNA Repair; DNA Repair Gene; DNA Repair Pathway; DNA-dependent protein kinase; Data Analyses; Data Collection; Environment; Eukaryotic Cell; Exclusion; Fluorescence; Goals; Gold; Individual; Institutes; Institution; Label; Laboratories; Ligands; Light; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Methods; Microscopy; Modality; Modeling; Molecular; Molecular Conformation; Mutation; New York; Nuclear; Optics; Patients; Phase Transition; Process; Proteins; Publications; RNA; Repair Complex; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Scientist; Signal Transduction; Source; Structure; Synchrotrons; System; Technology; Testing; Variant; Visit; Work; X-Ray Crystallography; X-Ray Tomography; activation product; beamline; crosslink; design; empowered; flexibility; genome integrity; imaging modality; member; photonics; programs; protein complex; protein purification; reconstitution; response; screening; sensor; single-molecule FRET; small molecule; structural biology

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.

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 和保持基因组完整性的组件。然而。 DR 蛋白和复合物具有编码灵活性 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 具有挑战性的。与 SBDR 研究人员合作，结构细胞生物学 (SCB) 核心 Core 2 克服了这些问题 通过提供尖端结构生物学技术的机会和专业知识来应对极端挑战 需要探测 DR 蛋白和复合物，但个别研究机构不具备。 目标 1. 确定结构、构象和组装状态。 SCB 核心，以国家为中心 同步加速器设施，将提供大分子晶体学 (MX) 和小角度 X 射线散射 (SAXS) 为 SBDR 成员提供数据收集和光束线科学家级专业知识。 MX 提供高分辨率原子 结构，而 SAXS 可以快速评估灵活性、解决方案构象和组件。 SCB萨克斯 包括通过 GOLD-SAXS 筛选配体或缓冲液以及评估 DNA 弯曲。 目标 2. 检测综合结构生物学的折叠和构象。大型复合物的分析 通过兆道尔顿结构质谱 (MS) 和经过 SAXS 验证的结构预测的项目将 自己提供可检验的假设，或者将加速各个研究所的 CryoEM 研究。 目标 3. 通过同步加速器软 X 射线定义重构系统和细胞中的分子结构 重建相变和细胞的断层扫描 (SXT) 以及超分辨率荧光 DR 组件的定量测量，否则会出现问题 传统的结构方法。 SCB 核心以先进光源 (ALS) 同步加速器为中心，其中三个光束线（12.3.1、 8.3.1 和国家 X 射线断层扫描中心/NCXT）将提供 SAXS、MX 和 SXT 数据收集 SBDR 成员。为了获得额外的结构技术和能力，SCB 设有分支机构 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 并以典型实验室和光束线的方式有效地解决项目结构和机械目标 访问不能或不做。这些 SCB 模式共同允许 SBDR-5 询问动态 DR 从原子到细胞尺度的机器。 SCB 核心的广泛、长期目标是支持 SBDR 确定序列变异影响的项目、剖析多个功能的方法以及 灵活、动态的灾难恢复机制发挥作用的机制。