Structure-Function of Nucleo-Cytoplasmic Communication

核-细胞质通讯的结构-功能

• 批准号: 10793672
• 负责人: Thomas Schwartz
• 金额: $ 9.28万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10793672
• 关键词: Address; Cardiac; Cardiomyopathies; Cell Nucleus; Cell physiology; Cells; Communication; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Cytoskeleton; Drug Design; Elements; Emery-Dreifuss Muscular Dystrophy; Eukaryotic Cell; Functional disorder; Genetic Materials; Genetic Transcription; Goals; Human; Individual; Life; Malignant Neoplasms; Maps; Membrane; Methods; Molecular; Muscular Dystrophies; Myopathy; Nuclear Envelope; Nuclear Pore Complex; Organelles; Positioning Attribute; Premature aging syndrome; Primary Dystonias; Process; Productivity; Proteins; Resolution; SWP29; Structure; Technology; Translations; X-Ray Crystallography; grasp; human disease; innovation; mechanotransduction; protein complex; skeletal; success

PROJECT SUMMARY / ABSTRACT Eukaryotic cells are defined by their organelles, membrane-enclosed compartments in which specific cellular processes are carried out. The nucleus is the largest organelle, contains all genetic material, and enables separation of gene transcription from protein translation. As the nuclear envelope (NE) serves as a tight barrier enclosing the nucleus, the cell requires machinery to establish and control nucleo-cytoplasmic communication. There are two principally different components to this machinery. Nuclear pore complexes (NPCs) serve as the main conduit for molecular exchange across the NE, while universally conserved linker of nucleo- and cytoskeleton (LINC) complexes serve as physical tethers across the NE. LINCs are necessary for positioning the nucleus and for mechano-sensing in a diverse set of circumstances. Dysfunction of the machinery is at the core of important human diseases, including skeletal and cardiac myopathies, premature aging, and cancer. Our goal is to understand the structure of the protein complexes involved in nucleo-cytoplasmic communication at high (atomic) resolution. Such information helps to identify and separate the myriad functions this machinery carries out and that we are still only beginning to fully grasp. High resolution information further provides the basis for structure-guided drug design to interfere with the salient human diseases, such as Emery-Dreifuss Muscular Dystrophy (EDMD) and Primary Dystonia, which are still not cured. The structural characterization of the NPC and the LINC complex are challenging, because of the size and complexity of these multi-MDa assemblies. Over the past 15 years, we have made significant advances on both problems. For the NPC, we have chosen a highly productive bottom-up approach, in which we characterized multi-subunit complexes predominantly by X-ray crystallography, the building blocks of the massive, 40-100 MDa NPC. Those structures have now been used in combination with cryo-electron tomographic (cryo-ET) maps of assembled NPCs to generate composite structures that can tentatively position the roughly 500 individual proteins within one NPC. For the LINC complex, we solved the universally conserved core component and have started to untangle the diverse network of its components, the Sad1/UNC-84 (SUN) and Klarsicht/ANC1/Syne-Homology (KASH) proteins. Going forward, the challenge is the structural characterization of large and dynamic assemblies, which is true for both, the NPC and the LINC complex, for the latter particularly when including the connection to the nucleo- and cytoskeletal components. The dramatic advances in cryo-electron microscopy (cryo-EM) over the recent past make this technology particularly important for our studies. We anticipate combining X-ray crystallography and cryo-EM for studying the most relevant structures going forward. The success of this will depend upon innovative methods to address the particular challenges that come with each project. We have repeatedly shown how to successfully approach such challenges and have devised methods to meet them.

项目摘要 /摘要 真核细胞由它们的细胞器，膜封闭的室定义，其中特定的细胞在其中 进行过程。核是最大的细胞器，包含所有遗传物质，并可以 基因转录与蛋白质翻译的分离。由于核信封（NE）充当紧密的障碍 封闭细胞核，该细胞需要机械才能建立和控制核总质质通信。 该机械有两个主要不同的组件。核孔复合物（NPC）作为 NE跨NE的分子交换的主要管道，而核和核和核的连接器 细胞骨架（LINC）复合物是整个NE的物理系数。肾脏对于定位是必要的 在各种情况下，核和机械感应。机械功能障碍在 重要的人类疾病的核心，包括骨骼和心脏肌病，过早衰老和癌症。我们的 目的是了解涉及核总质通信的蛋白质复合物的结构 高（原子）分辨率。此类信息有助于识别和分离这种机械的无数功能 执行，我们仍然刚刚开始完全掌握。高分辨率信息进一步提供了 结构引导的药物设计的基础，以干扰显着的人类疾病，例如Emery-Dreifuss 肌肉营养不良（EDMD）和原发性肌张力障碍仍未治愈。结构表征 由于这些多MDA的大小和复杂性 集会。在过去的15年中，我们在这两个问题上都取得了重大进步。对于NPC，我们 已经选择了一种高效的自下而上的方法，在这种方法中，我们表征了多生产配合物 主要由X射线晶体学，巨大的40-100 MDA NPC的构件。那些结构 现在已与组装NPC的冷冻电子层析成像（Cryo-ET）结合使用 生成可以在一个NPC内暂时将大约500个单个蛋白质定位的复合结构。 对于LINC综合体，我们解决了普遍保守的核心组件，并开始解开 其组件的各种网络，SAD1/UNC-84（SUN）和Klarsicht/anc1/syne-andomology（KASH） 蛋白质。展望未来，挑战是大型和动态组件的结构表征， NPC和LINC综合体都是正确的，对于后者，尤其是在包括与 核和细胞骨架成分。在 最近的过去使这项技术对我们的研究特别重要。我们预计结合X射线 晶体学和冷冻EM用于研究未来最相关的结构。成功的成功 依靠创新方法来应对每个项目带来的特定挑战。我们有 反复展示了如何成功应对此类挑战并设计了满足它们的方法。

项目成果

Solving the nuclear pore puzzle.

解决核孔之谜。

• DOI: 10.1126/science.abq4792
• 发表时间: 2022
• 期刊: Science (New York, N.Y.)
• 作者: Schwartz,ThomasU

The cellular environment shapes the nuclear pore complex architecture.

• DOI: 10.1038/s41586-021-03985-3
• 发表时间: 2021-10
• 期刊: Nature
• 影响因子: 64.8
• 作者: Schuller AP;Wojtynek M;Mankus D;Tatli M;Kronenberg-Tenga R;Regmi SG;Dip PV;Lytton-Jean AKR;Brignole EJ;Dasso M;Weis K;Medalia O;Schwartz TU
• 通讯作者: Schwartz TU