The role of microtubule plus-end capture at cortical sites in the assembly of apico-basal microtubules in polarised epithelial cells.

皮质位点微管正端捕获在极化上皮细胞中顶端-基底微管组装中的作用。

• 批准号: BB/D012201/1
• 负责人: Mette Mogensen
• 金额: $ 32.42万
• 依托单位: University of East Anglia
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD012201%2F1
• 关键词: role; microtubule; plus; end; capture

Microtubules are tubular structures, which are important for many cellular functions including the transport of vesicles and molecules within cells. It is therefore very important that the microtubules are assembled and positioned correctly within cells. The specific pattern that microtubules form within cells vary depending on cell type and function. Most animal cells have a radial array of microtubules anchored at a centrally located structure called the centrosome. In these cells the centrosome's job is to start off the assembly of the microtubules and to keep them tightly bound to it. However, many specialised cells like polarised epithelial cells found in the gut and kidneys arrange their microtubules in a different way. Here most of the microtubules are anchored at the cell apex away from the centrosome and run towards the base forming a so called apico-basal array. Our recent findings suggest that the apico-basal microtubules are assembled at the centrosome, released and moved to apical sites where they are captured and anchored. The aim of this project is to find out whether the microtubules first grow out from the centrosome, make contact with the cell cortex, are released from the centrosome and then move downward by the pulling action of dynein motor proteins located at the cortex. The end result would be that one end (plus-end) of the microtubules is pulled down to the cell base while the other end (minus-end) becomes anchored at the cell apex. We want to find out whether proteins such as EB1 and CLIP-170, which stick to the growing end of microtubules, are important for the capture by dynein or other proteins at the cortex. It is very important to know if for example CLIP-170 is vital for the normal assembly of the microtubule in real life. We will therefore analyse tissue from the inner ear (cochlea) isolated from mice, which do not produce CLIP-170. Finally, we would also like to know whether thin filaments known as actin help the microtubules to get to their destination. We will use special fluorescent (glowing) molecules called GFPs linked to proteins so that the microtubules or their plus-ends glow and we will make movies of their movements over time and look at the localisation of these important proteins using fluorescent dyes and specialised microscopes. Microtubules are clearly vital for the normal function of a cell and we need to establish how they are organised and controlled before we can unravel the causes and consequences of many diseases.

微管是一种管状结构，它对许多细胞功能都很重要，包括细胞内小泡和分子的运输。因此，微管在细胞内正确组装和定位非常重要。微管在细胞内形成的特定模式取决于细胞类型和功能。大多数动物细胞都有一个放射状排列的微管，它们锚定在一个位于中心的结构上，称为中心体。在这些细胞中，中心体的工作是启动微管的组装，并使它们紧密结合在中心体上。然而，许多特化细胞，如在肠道和肾脏中发现的极化上皮细胞，以不同的方式排列微管。在这里，大多数微管锚定在远离中心体的细胞顶端，并向基部延伸，形成所谓的顶端-基部阵列。我们最近的研究结果表明，顶端-基底微管组装在中心体，释放和移动到顶端的网站，他们被捕获和锚定。该项目的目的是发现微管是否首先从中心体生长出来，与细胞皮层接触，从中心体释放，然后通过位于皮层的动力蛋白马达蛋白的拉动作用向下移动。最终的结果是微管的一端（正端）被拉到细胞基部，而另一端（负端）锚定在细胞顶端。我们想知道，粘附在微管生长端的EB 1和CLIP-170等蛋白质，是否对皮层的动力蛋白或其他蛋白质的捕获很重要。例如，了解CLIP-170是否对真实的生活中微管的正常组装至关重要是非常重要的。因此，我们将分析从小鼠分离的内耳（耳蜗）组织，这些组织不产生CLIP-170。最后，我们还想知道，被称为肌动蛋白的细丝是否有助于微管到达目的地。我们将使用特殊的荧光（发光）分子称为GFP连接到蛋白质，使微管或其加端发光，我们将制作它们随时间推移的运动电影，并使用荧光染料和专业显微镜观察这些重要蛋白质的定位。微管对于细胞的正常功能显然是至关重要的，我们需要确定它们是如何组织和控制的，然后才能解开许多疾病的原因和后果。

项目成果

Reorganization of centrosomal marker proteins coincides with epithelial cell differentiation in the vertebrate lens.

中心体标记蛋白的重组与脊椎动物晶状体中上皮细胞的分化同时发生。

• DOI: 10.1016/j.exer.2007.07.022
• 发表时间: 2007
• 期刊: Experimental eye research
• 影响因子: 3.4
• 作者: Dahm R

Overly long centrioles and defective cell division upon excess of the SAS-4-related protein CPAP.

SAS-4 相关蛋白 CPAP 过量时，中心粒过长，细胞分裂有缺陷。

• DOI: 10.1016/j.cub.2009.05.018
• 发表时间: 2009-06-23
• 期刊: CURRENT BIOLOGY
• 影响因子: 9.2
• 作者: Kohlmaier, Gregor;Loncarek, Jadranka;Meng, Xing;McEwen, Bruce F.;Mogensen, Mette M.;Spektor, Alexander;Dynlacht, Brian D.;Khodjakov, Alexey;Goenczy, Pierre
• 通讯作者: Goenczy, Pierre