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Atomic Structure of the Nuclear Pore Complex

核孔复合体的原子结构

基本信息

批准号：
10242216
负责人：
Andre Hoelz
金额：
$ 37.73万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-05 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10242216
关键词：
Accounting Address Anatomy Architecture Binding Biochemical Biogenesis Biological C-terminal Caliber Cell Cycle Cell Nucleus Cell physiology Cells Characteristics Complex Cryo-electron tomography Crystallization Cytoplasmic Filaments Disease Eukaryotic Cell Event Evolution Functional disorder Gene Expression Regulation Genetic Transcription Grant Health Human Inherited Integral Membrane Protein Knowledge Life Link Maps Membrane Methodology Mitosis Molecular N-terminal Nature Nuclear Nuclear Envelope Nuclear Import Nuclear Pore Complex Nuclear Pore Complex Proteins Nuclear Structure Nucleic Acids Organelles Outcome Physiological Post-Translational Protein Processing Proteins Recombinants Research Role Structure Validation Work Yeasts flexibility genetic information human disease in vivo insight leukemia mRNA Export macromolecular assembly macromolecule molecular mass nucleocytoplasmic transport protein folding public health relevance reconstitution reconstruction scaffold success

项目摘要

MODIFIED ABSTRACT The enclosure of genetic information in the nucleus is one of the great hallmarks of evolution, but creates the necessity for dedicated portals through which folded proteins and protein/nucleic acid complexes can cross the nuclear envelope (NE). The nuclear pore complex (NPC), a cylindrical supramolecular structure embedded in circular pores permeating the NE, is the sole gateway for passage through the NE and can accomplish the selective bidirectional transport of macromolecules of up to ~40 nm in diameter at a rate of several hundred events per second. Beyond its primary role in nucleocytoplasmic transport, the NPC also contributes to additional modes of gene regulation for example through direct interaction with the transcription and mRNA export machineries. The NPC thus represents an essential organelle for all eukaryotic life and, accordingly, NPC dysfunction has been associated with various forms of human disease. Architecturally, the NPC consists of a central symmetric core to which asymmetric components called cytoplasmic filaments and nuclear basket are attached. The NPC is built from ~34 different proteins termed nucleoporins that are each present in multiple copies such that the entire assembly reaches the extraordinary mass of ~110 MDa in humans. Nucleoporins are organized into distinct subcomplexes which constitute physiological building blocks of the intact NPC in vivo. To determine the atomic architecture of the NPC, my group has been pursuing a divide-and-conquer approach, in which we have mapped nucleoporin interactions, reconstituted recombinant nucleoporin complexes and determined their crystal structures to be fit into cryo-electron tomographic reconstructions of the intact NPC. In this way, we achieved a near-atomic composite structure of the ~60MDa human NPC symmetric core in the previous grant period. Building on this progress, we now propose to expand our structural characterization to still unresolved parts of the NPC and to use our already gained knowledge to address fundamental NPC-associated cell biological questions. Specifically, we plan to elucidate the molecular interactions in the NPC’s inner ring that are essential for the formation of its central transport channel, and between the symmetric core and transmembrane NPC components that are essential for NPC anchoring in the NE pores. The outcome of the proposed research is expected to greatly increase our understanding of the molecular mechanisms by which the NPC regulates nucleocytoplasmic transport and associated cellular processes, while simultaneously creating a mechanistic basis for currently untreatable “nup diseases.” Furthermore, the methodologies developed herein will serve as a paradigm for the characterization of other essential cellular mega-assemblies as large, flexible and complex as the NPC whose functional mechanisms have remained elusive due to lack of structural insight.

修改后的摘要