Patterning acentrosomal microtubule arrays

总结 微管细胞骨架支持细胞分裂、细胞形态和细胞内货物运输。 虽然中心体是分裂细胞中微管成核的主要场所，但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 以实现聚合物数量、长度和动力学的特定模式， 细胞功能，通常贯穿有机体的整个生命周期。我的实验室研究 C.无中心体微管及其对货物运输的影响优雅我们开发了成像工具， 这些算法允许对体内微管组织进行前所未有的分析， 货物运输的实时成像。我们进行了公正的筛选，以发现新的微管调节剂 并正在使用遗传学，成像和生物化学方法来了解其机制。同时，我们 研究微管阵列模式的生物学意义，通过检查这些影响， 长距离细胞内运输的调节剂。该提案详细介绍了我们的实验性 系统，屏幕的设计和实现，以及选定调节器的初步特性。然后它 概述了我们未来五年的主要目标：完成筛选并阐明 建立无中心体阵列结构。这些研究将确定稳态阵列架构 从单个聚合物成核和动力学的控制中出现，以及它如何适应 专门的细胞微管支持基本的生物过程，如细胞迁移，极化 和货物运输。因此，我们的工作将产生重大影响：它将确定新的监管机构， 无中心体阵列的构建模块，它将决定它们模式化的机制。 细胞骨架和调节运输。细胞骨架缺陷与许多疾病的关系表明 从长远来看，我们的研究将有助于阐明细胞功能障碍的机制， 在疾病期间。

Summary 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.