NPC ASSEMBLY · The Nuclear Pore Complex: Assembly Required
6РП — Действия „Мария Кюри“
- Период
- 2006-09-01 → 2008-08-31
- Финансиране от ЕС
- 80 000 €
- Участници
- 1
- Схема
- IRG
Линиите свързват координатора с партньорите.
Накратко на български
Ядрените пори контролират движението на вещества между ядрото и останалата част на клетката, като в случая се изучава как се сглобяват тези структури. Разбирането на този процес помага при биомедицинските изследвания и бъдещото създаване на лекарства.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - NPC ASSEMBLY (The Nuclear Pore Complex: Assembly Required)
The genome of human cells, plant, animals, and even some unicellular organisms is segregated from the rest of the cell and kept inside the nucleus. The nuclear pore complex (NPC) mediates all traffic between the nucleus and the rest of the cell. The regulation of this traffic affects every aspect of gene expression and a multitude of other cellular processes. The assembly and function of the NPC are therefore a major topic for biomedical research as well as future drug design. We are using a cell-free system derived from frog egg extracts to assemble functional nuclei in the test-tube and to study the mechanism of NPC assembly. To gain a better understanding of this process, we set out to molecularly define early steps in nuclear pore assembly, beginning with chromatin binding. The order of recruitment of the first few building blocks, including two key integral pore-membrane proteins, was established. We find that most of the soluble nuclear pore subunits are added only after the recruitment of these key membrane components. These findings strongly support one of the two conflicting mechanistic models that have been suggested thus far to explain NPC assembly. In addition, we have isolated native soluble nuclear pore subunits from egg extracts and we continued to investigate the regulatory role of the prototypic import receptor, importin, in NPC assembly. Overall, our results provide a detailed view of the early stages of NPC assembly, as well as the basis for mapping the exact molecular interfaces that determine this process.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The nuclear pore complex (NPC) mediates all traffic between the nucleus and cytoplasm and serves as a central node of cellular communication. Despite the recent identification of the full complement of nuclear pore proteins, much of the molecular details of NPC structure and its mechanism of action, remain unknown. Moreover, the precise molecular pathway for NPC biogenesis is largely undefined. Here, I present a comprehensive approach for dissecting the mechanism of NPC assembly, using in vitro nuclear reconstitution in Xenopus egg extracts. My working hypothesis is that NPC assembly begins with the formation of fused double nuclear membranes. In this model, an initial fusion event between the inner and outer nuclear membranes triggers the assembly process, and soluble nucleoporins are then added in a sequential manner, to form the complete structure.A prediction of this model is that pore-membrane protein(s) should play a vital role at the early stages of assembly. To test this prediction we will use morphological analysis and immunolocalization of the pore-membrane proteins by transmission electron microscopy. Aborted assembly intermediates will be isolated and used in combination with fractionated cytosol to identify subsequent steps in the process. Newly identified assembly intermediates will be characterize by fluorescence and electron microscopy. The overall goal is to reach a detailed step-by-step model of NPC assembly. This will promote our understanding of the biogenesis and function of one of the largest supramolecular assemblies in eukaryotic cells, and the gateway to the genome.
Оригинален текст от CORDIS (на английски).
Участници
- TECHNION - ISRAEL INSTITUTE OF TECHNOLOGY · HAIFAКоординаторИзраел
Връзки
Данни: CORDIS, © Европейски съюз