Patterning acentrosomal microtubule arrays

摘要 微管细胞骨架支持细胞分裂、细胞形态和细胞内货物运输。而 中心体是分裂细胞中微管成核的主要场所，许多分化的细胞具有 无中心体微管阵列突出的实例包括生殖系细胞、植物表皮、上皮细胞和细胞外基质。 神经元为了理解细胞分化，了解无中心体阵列结构是如何设置的至关重要 以获得特定的聚合物数量、长度和动力学模式， 功能，通常贯穿于有机体的整个生命周期。 本实验室主要研究C.优雅 我们开发了成像工具和算法，可以对微管进行前所未有的分析。 组织在体内，并与货物运输的实时成像兼容。我们进行了公正的筛选， 发现新的微管调节器，并正在使用遗传学，成像和生物化学方法来了解 他们的机制。与此同时，我们正在研究微管阵列模式的生物学意义， 检查这些调节剂对长距离细胞内转运的影响。该提案详细说明了 建立了我们的实验系统，设计并实现了筛选，并初步 选择监管机构的特征。然后它概述了我们未来五年的主要目标：完成屏幕 并阐明建立无中心体阵列架构的机制。这些研究将决定 稳态阵列结构如何从单个聚合物成核和动力学的控制中出现， 它是如何适应特殊细胞的功能的。 微管支持细胞迁移、极化和货物运输等基本生物过程。 因此，我们的工作将产生重大影响：它将确定新的监管机构， acentrosomal阵列，它将确定其模式的细胞骨架和调节机制， 运输在许多疾病中涉及细胞骨架缺陷表明，从长远来看， 研究将有助于阐明疾病期间发生的细胞功能障碍的机制。

Abstract The microtubule cytoskeleton supports cell-division, cellular morphology and intracellular cargo transport. While the centrosome is a major site of microtubule nucleation in dividing cells, many differentiated cells harbor acentrosomal microtubule arrays. Prominent examples include germline cells, plant epidermis, epithelia and neurons. To understand cellular differentiation, it is crucial to learn how acentrosomal array architecture is set up to achieve a specific pattern of polymer numbers, length and dynamics that would support specialized cellular functions, often throughout the life of an organism. My laboratory studies the patterning of acentrosomal microtubules and its effect on cargo transport in C. elegans. We developed imaging tools and algorithms that allow an unprecedented level of analysis of microtubule organization in vivo and are compatible with live-imaging of cargo transport. We conducted unbiased screens to uncover novel microtubule regulators and are using genetics, imaging, and biochemical methods to understand their mechanisms. In parallel, we are investigating the biological significance of microtubule array patterns by examining the effects of these regulators on long-range intracellular transport. This proposal details the establishment of our experimental system, design and implementation of the screen, and preliminary characterization of select regulators. It then outlines our main goals for the next five years: completing the screen and elucidating the mechanisms that establish acentrosomal array architecture. These studies will determine how steady-state array architecture emerges from the control of single polymer nucleation and dynamics and how it is adapted to the function of specialized cells. Microtubules support fundamental biological processes such as cell migration, polarization and cargo transport. Hence, our work will have a significant impact: It will identify novel regulators that arrange the building blocks of acentrosomal arrays and it will determine the mechanisms by which they pattern the cytoskeleton and regulate transport. The involvement of cytoskeletal defects in numerous disorders suggests that in the long-range, our studies will help to shed light on mechanisms of cellular dysfunction that occurs during disease.