A versatile platform for reconstructing the spatial organization of intracellular signaling during cell-division

用于重建细胞分裂过程中细胞内信号传导空间组织的多功能平台

基本信息

批准号：
9348934
负责人：
Radhika Subramanian
金额：
$ 256.5万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2022-08-31
项目状态：
已结题

项目摘要

ABSTRACT A central question in cell biology is how spatial information on the micron-length scale is encoded within the cellular cytoplasm. This is essential for maintaining genome integrity during mitosis where the cell-cleavage plane must be precisely positioned at the center of a ~ 10-15 um long cell during cytokinesis. It is now well established that the microtubule cytoskeleton plays a critical role in this process by forming a specialized structure proximal to the site of cell cleavage. This structure, known as the spindle midzone, acts as a micron scale `mark' for the cell center. These micron-scale marks are then `read' by signaling proteins that generate self- organizing activity patterns that further precisely define the site of cell-cleavage. Subsequently this information is transmitted to the cell cortex for the organization of the contractile ring. We now have a near complete parts list of the proteins that regulate these processes. However, the molecular mechanism by which the midzone templates the formation of spatial activity patterns of regulatory proteins within the cytoplasm and the cell cortex is starkly incomplete. Here we will develop in vitro reconstitution assays using microtubules, reconstituted bilayer, singe molecule FRET sensors and DNA origami-based nanotemplates to decipher how spatial organization of signaling activity is achieved on microtubule templates and how these reactions are organized at membrane-microtubule interface. These studies will provide insights into a conserved signaling cascade that is critical for cell division and its mis-regulation in human cancer. Importantly, the innovative methodologies developed here will be widely applicable to dissecting other signaling mechanisms by which spatial activity patterns are encoded and deciphered in the cellular cytoplasm during cell division, growth and development.

抽象的细胞生物学的一个中心问题是微米长度尺度的空间信息是如何表达的编码于细胞质内。这对于维持基因组完整性至关重要有丝分裂，其中细胞分裂平面必须精确定位在 ~ 10-15 的中心胞质分裂期间的长细胞。现在已经确定微管细胞骨架通过在细胞位点附近形成特殊结构，在此过程中发挥关键作用乳沟。这种结构被称为主轴中区，充当微米级“标记” 细胞中心。然后，这些微米级标记被信号蛋白“读取”，这些信号蛋白会产生自我标记。组织活动模式，进一步精确定义细胞分裂的位点。随后该信息被传输到细胞皮层以组织收缩环。我们现在已经有了一份几乎完整的调节这些过程的蛋白质的零件清单。然而，中区模板形成空间活动模式的分子机制细胞质和细胞皮层内的调节蛋白的结构完全不完整。在这里我们将使用微管、重构双层、单分子开发体外重构测定 FRET 传感器和基于 DNA 折纸的纳米模板可破译 DNA 的空间组织方式信号活动是在微管模板上实现的以及这些反应是如何组织的在膜-微管界面。这些研究将为保守的信号级联对于细胞分裂及其在人类癌症中的错误调节至关重要。重要的是，这里开发的创新方法将广泛应用于解剖空间活动模式被编码和破译的其他信号机制细胞分裂、生长和发育过程中的细胞质。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Atomic force microscopy reveals distinct protofilament-scale structural dynamics in depolymerizing microtubule arrays.

DOI：
10.1073/pnas.2115708119
发表时间：
2022-02-01
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Wijeratne SS;Marchan MF;Tresback JS;Subramanian R
通讯作者：
Subramanian R

Active Microphase Separation in Mixtures of Microtubules and Tip-Accumulating Molecular Motors