Molecular mechanisms of kinesin-5s in fungal mitosis

这项研究的目的是发现细胞复制是如何控制的，以及它是如何被阻止的。我们将把我们的研究重点放在真菌中的细胞复制上，这既是因为它们是研究复制的优秀和完善的模式生物，也是因为真菌疾病在医学，环境和经济上都很重要。特别是，我们想研究感染玉米并导致玉米黑穗病的真菌中的细胞复制机制。这种作物病害对全球粮食安全构成重大威胁，特别是因为对现有杀真菌剂出现了抗药性。通过了解玉米黑粉菌如何复制，我们希望首先提供对细胞复制机制的一般了解。第二，我们希望发现真菌细胞复制的独特特征，因为这些知识有望帮助开发新的杀菌剂。就像我们的身体有一个为我们提供支撑和力量的骨架一样，真菌的细胞也有一个骨架-称为细胞骨架-它也提供支撑和结构。细胞骨架参与了真菌生活的许多重要方面，包括细胞运输，建筑和复制。研究细胞骨架很重要，不仅可以让我们了解健康细胞的工作原理，还可以让我们用药物特异性地靶向致病生物的细胞骨架，杀死这些生物并预防疾病。特别是，该项目将重点关注细胞骨架的一部分，称为微管。这些是长的圆柱形结构，就像分子运输马达沿着携带细胞货物的轨道。我们将要研究的马达被称为驱动蛋白，有许多不同的类型。在这个项目中，我们将研究驱动蛋白5型，这对精确的细胞复制很重要。我们想知道真菌驱动蛋白-5在复制过程中是如何利用细胞燃料沿着沿着移动的，以及这种活动是如何被阻断的。伯克贝克研究小组的工作将涉及研究细胞骨架的三维结构，因为知道细胞骨架的样子将有助于我们了解它在真菌本身中是如何工作的。我们将使用一个非常强大的显微镜-电子显微镜-拍摄单个细胞骨架分子的照片，然后使用计算分析联合收割机将这些照片组合起来，计算出它们的三维形状。该项目的一个重要方面是，我们还将与埃克塞特大学的真菌细胞生物学专家合作，研究驱动蛋白-5在活玉米黑粉菌中的功能。与许多其他真菌，这是非常小的，玉米黑粉菌的细胞是相对较大的（10微米）。这意味着可以使用光学显微镜详细研究单个细胞的细胞复制，因此我们的合作者将能够可视化活真菌中驱动蛋白-5s的活性，并检查阻断其活性对真菌存活的影响。初步分析表明，来自真菌的驱动蛋白-5s与来自其他生物体（包括人类）的驱动蛋白-5s不同。这意味着我们可能能够找到可以阻断真菌驱动蛋白-5而不是人类驱动蛋白-5的药物，并且这些药物可能非常有希望作为新的杀真菌剂开发。研究真菌驱动蛋白-5s的结构和功能将使我们能够研究这一想法。

The purpose of this research is to discover how cell replication is controlled and how it might be blocked. We will focus our studies on cell replication in fungi, both because they are excellent and well-established model organisms for studying replication, and because fungal diseases are medically, environmentally and economically important. In particular, we want to investigate the cell replication machinery in a fungus that infects corn and causes the disease corn smut. This crop disease poses a major threat to global food security, particularly because of the emergence of resistance to currently available fungicides. By understanding how the corn smut fungus replicates, we hope to first, provide general insight into the mechanisms of cell replication. Secondly, we hope to uncover unique features of fungus-specific cell replication because this knowledge promises to help in the development of novel fungicides.In the same way as our bodies have a skeleton that provides us with support and strength, the cells of fungi have a skeleton - called the cytoskeleton - which also provides support and structure. The cytoskeleton is involved in many important aspects of the life of fungi, including cellular transport, architecture and replication. Studying the cytoskeleton is important both so we can understand how healthy cells work, but also so we can specifically target the cytoskeleton of pathogenic organisms with drugs that kill these organisms and prevent disease.In particular, this project will focus on a part of the cytoskeleton called microtubules. These are long cylindrical structures that act like tracks along which molecular transport motors carry cellular cargo. The motors that we will study are called kinesins and there are many different types. In this project, we will be studying a kinesin type-5, which is important for accurate cell replication. We want to know how fungal kinesin-5s use cellular fuel to move along microtubules during replication and how this activity might be blocked. The work by the Birkbeck research team will involve studying the three-dimensional structure of the cytoskeleton, because knowing what the cytoskeleton looks like will contribute to our understanding of how it works in the fungus itself. We will use a very powerful microscope - an electron microscope - to take pictures of individual cytoskeleton molecules and then use computational analysis to combine these pictures and calculate their three-dimensional shape. A powerful aspect of the proposed project is that we will also study the function of kinesin-5 in live corn smut fungus, in collaboration with experts in fungal cell biology at the University of Exeter. Unlike many other fungi, which are very small, the cells of the corn smut fungus are relatively large (10um). This means that cell replication of individual cells can be studied in detail using light microscopy, so that our collaborators will be able to visualise the activity of kinesin-5s in the living fungus and examine the effects of blocking its activity on fungal survival.Initial analysis suggests that the kinesin-5s from fungi are different from kinesin-5s from other organisms, including humans. This means that we might be able to find drugs that can block fungal kinesin-5 and not human kinesin-5, and such drugs could be very promising for development as new fungicides. Studying the structure and function of the fungal kinesin-5s will allow us to investigate this idea.