Cellular function and regulation of RCC1 isoforms

RCC1 亚型的细胞功能和调节

基本信息

批准号：
BB/G001480/1
负责人：
Paul Clarke
金额：
$ 40.99万
依托单位：
University of Dundee
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2008
资助国家：
英国
起止时间：
2008 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FG001480%2F1
关键词：
Cellular function regulation RCC1 isoforms

项目摘要

Living organisms are made of cells. The genes that control all the functions of each cell are made of DNA and are collected in several chromosomes that are contained in the cell nucleus. Communication between the nucleus and the rest of the cell (the cytoplasm) is important for the production of proteins encoded by the genes. The interaction between the nucleus and the rest of the cell may be important for the spatial organisation of the cell, particularly during cell division, when the nucleus is dramatically reorganised. In animal and plant cells, the nuclear envelope, a membrane structure that surrounds the chromosomes, breaks down during cell division, and pairs of chromosomes become arranged on a structure called the spindle. One of each pair of chromosomes is separated to each end of the spindle, then this structure breaks down and the cell is pinched in two, leaving one set of chromosomes to form a new nucleus in each new cell. During cell division, the spatial orientation of the chromosomes and the spindle relative to the rest of the cell determines the plane of cell division. This is important in tissues, where cells are arranged relative to each other and their intracellular structures are polarised, that is arranged in a particular direction. A protein called Ran is important in the communication between the nucleus and the rest of the cell. The active form of Ran is made at chromosomes by another protein, RCC1. Active Ran directs the movement of other proteins and large molecules between the nucleus and cytoplasm. It also directs spindle assembly during mitosis and nuclear envelope formation. We have found that RCC1 exists in cells in at least three different forms (isoforms) that we think may have different functions and may be important when cells actively proliferate during development and in cancer. We will study how these different RCC1 isoforms work, using extracts of human cells and frog eggs which can reproduce complicated processes like spindle assembly and nuclear formation in a test tube. We will investigate the protein complexes made by RCC1 isoforms by a technique called mass spectrometry. We will also use advanced techniques for microscopy to study how RCC1 isoforms work in living cells that are grown on a Petri dish. To do this, we will make RCC1 isoforms in the cells coupled to a naturally fluorescent protein from jellyfish. This work will enable us understand how RCC1 isoforms work and how complicated processes in cells are coordinated by communication between the nucleus and cytoplasm. This work may help us to understand how to control cell division and tissue growth, and how they can go wrong in diseases.

活生物体是由细胞制成的。控制每个细胞的所有功能的基因都是由DNA制成的，并收集在包含在细胞核中的几个染色体中。细胞核与细胞其余部分（细胞质）之间的通信对于由基因编码的蛋白质的产生很重要。细胞核与细胞其余部分之间的相互作用对于细胞的空间组织，尤其是在细胞分裂期间，当细胞核被大大重组时。在动物和植物细胞中，核包膜是围绕染色体的膜结构，在细胞分裂过程中分解，染色体对成对被排列在称为纺锤体的结构上。每对染色体中的一个分开与主轴的每一端分开，然后将细胞分解为两部分，并将一组染色体夹在每个新细胞中。在细胞分裂期间，染色体的空间取向和纺锤体相对于细胞的其余部分决定了细胞分裂的平面。这在组织中很重要，在组织中，细胞相对于彼此排列，其细胞内结构是极化的，该结构是在特定方向上排列的。一种称为RAN的蛋白质在细胞核与细胞其余部分之间的通信中很重要。 RAN的活性形式是由另一种蛋白RCC1在染色体上制成的。活性引导其他蛋白质和细胞质之间的大分子的运动。它还指导有丝分裂和核包膜形成期间主轴组件。我们发现，RCC1在至少三种不同形式的细胞中存在，我们认为它们可能具有不同的功能，并且当细胞在发育和癌症中积极增殖时可能很重要。我们将使用人类细胞和青蛙卵的提取物研究这些不同的RCC1同工型如何起作用，这些提取物可以在试管中繁殖复杂的过程，例如主轴组装和核形成。我们将通过一种称为质谱法的技术研究RCC1同工型制成的蛋白质复合物。我们还将使用高级技术进行显微镜研究RCC1同工型如何在培养皿上生长的活细胞中起作用。为此，我们将在细胞中制作RCC1同工型，并从水母中耦合到天然荧光蛋白。这项工作将使我们能够理解RCC1同工型如何工作，以及细胞中细胞核与细胞质之间的通信协调的细胞中的复杂过程。这项工作可能有助于我们了解如何控制细胞分裂和组织生长，以及它们如何在疾病中出错。