The molecular roles of polycystin in cytokinesis

多囊蛋白在胞质分裂中的分子作用

• 批准号: 9813243
• 负责人: Qian Chen
• 金额: $ 44.41万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9813243
• 关键词: Affect; American; Animal Model; Attenuated; Autosomal Dominant Polycystic Kidney; C-terminal; Calcineurin; Calcium; Calcium Signaling; Cartoons; Cations; Cell Nucleus; Cell Proliferation; Cell Proliferation Regulation; Cell Separation; Cell division; Cells; Cellular biology; Coiled-Coil Domain; Confocal Microscopy; Cyst; Cytokinesis; Data; Daughter; Defect; Development; Environment; Failure; First Generation College Students; Fission Yeast; Fluorescence; Fluorescence Microscopy; Genetic Diseases; Homeostasis; Human; Human Genetics; Image Analysis; Individual; Integral Membrane Protein; Ion Channel; Ions; Isotonic Exercise; Kidney; Kidney Failure; Lead; Left; Light; Link; Lipid Binding; Liquid substance; Mediating; Methods; Microscopy; Molecular; Mutation; N-terminal; Osmolalities; PKD1 gene; Pathogenesis; Patients; Play; Proteins; Quantitative Microscopy; Regulation; Reporter; Research; Role; Signal Pathway; Site; Stress; Training; Universities; Work; Yeasts; constriction; daughter cell; genetic analysis; innovation; insight; loss of function mutation; mutant; novel; pressure; public health relevance; quantitative imaging; receptor; recruit; student training; tool; undergraduate research

SUMMARY Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common human genetic disorders, affecting more than 600,00 Americans. It is characterized by progressive development of liquid-filled cysts and increased cell proliferation in the kidney, often resulting in terminal renal failure. Despite of the discovery that mutations of two polycystin genes PKD1 and PKD2 account for all cases of ADPKD, it remains unclear why. We have uncovered a novel function of polycystin in cell division in fission yeast, that will shed new light on altered cell proliferation observed in ADPKD. We found that fission yeast polycystin Pkd2p localizes to the cell division site to regulate cytokinesis, the last stage of cell division. Here we propose to use this model organism to determine the molecular roles of polycystin in cytokinesis. Our central hypothesis is that Pkd2p regulates both the contractile ring and the calcium signaling during cytokinesis through its activity as an ion channel. Aim 1 To determine how Pkd2p regulates the contractile ring in cytokinesis. We found that the pkd2 mutation surprisingly leads to abnormal constriction of the ring and a large number of the mutant cells are under osmotic stress. We will first determine whether Pkd2p regulates the turgor pressure. We will then determine how Pkd2p regulates the assembly and the constriction of the contractile ring. We expect to confirm that Pkd2 modulates the assembly of the contractile ring by regulating the cellular osmolality. Aim 2 To determine how Pkd2p regulates the daughter cell separation in cytokinesis. Our preliminary data showed that Pkd2p plays an essential role in the separation of daughter cells. We propose to determine how Pkd2p regulates the calcium concentration in dividing cells. We will also examine how Pkd2p attenuates an essential signaling pathway that regulates the cell separation. We expect to confirm that Pkd2p regulates the calcium concentration in dividing cells and antagonizes the essential signaling pathway. Aim 3 To determine how different domains of Pkd2p protein work in cytokinesis. Pkd2p possesses three distinct domains with unknown functions. We propose to determine both localization and function of Pkd2p when these domains are removed individually. We expect to confirm that only two domains of Pkd2p are essential with each playing a separate role in cytokinesis. Despite years of research on polycystins, little is known about its function in cell division. Our surprising observation would provide novel insight into the regulation of cell proliferation by polycystins in the pathogenesis of ADPKD. We will use innovative microscopy method in our study. We propose to strongly promote undergraduate research to benefit the large number of first-generation college students at the University of Toledo and to train them to become immersed in quantitative cell biology research.

概括 常染色体显性多囊肾病（ADPKD）是最常见的人类遗传性疾病之一， 影响超过60万美国人。其特征是逐渐发展充满液体的囊肿和 肾脏细胞增殖增加，常常导致终末期肾衰竭。尽管发现 所有 ADPKD 病例均由两个多囊蛋白基因 PKD1 和 PKD2 的突变引起，但其原因尚不清楚。我们 发现了多囊蛋白在裂殖酵母细胞分裂中的一种新功能，这将为改变 ADPKD 中观察到的细胞增殖。我们发现裂殖酵母多囊蛋白 Pkd2p 定位于细胞分裂 调节胞质分裂（细胞分裂的最后阶段）的位点。在这里，我们建议使用这种模式生物 确定多囊蛋白在胞质分裂中的分子作用。我们的中心假设是 Pkd2p 调节 胞质分裂过程中收缩环和钙信号传导通过其作为离子通道的活性。 目标 1 确定 Pkd2p 如何调节胞质分裂中的收缩环。我们发现pkd2突变 令人惊讶的是，导致环异常收缩，并且大量突变细胞处于渗透状态 压力。我们首先确定 Pkd2p 是否调节膨胀压力。然后我们将确定 Pkd2p 如何 调节收缩环的组装和收缩。我们期望确认 Pkd2 的调节 通过调节细胞渗透压来组装收缩环。 目标 2 确定 Pkd2p 如何调节胞质分裂中的子细胞分离。我们的初步数据 表明 Pkd2p 在子细胞的分离中发挥重要作用。我们建议确定如何 Pkd2p 调节分裂细胞中的钙浓度。我们还将研究 Pkd2p 如何减弱 调节细胞分离的重要信号通路。我们期望确认 Pkd2p 监管 分裂细胞中的钙浓度并拮抗重要的信号传导途径。 目标 3 确定 Pkd2p 蛋白的不同结构域如何在胞质分裂中发挥作用。 Pkd2p 具有三个不同的 具有未知功能的域。我们建议在以下情况下确定 Pkd2p 的定位和功能： 域被单独删除。我们希望确认 Pkd2p 中只有两个域对于每个域都是必需的 在胞质分裂中发挥独立的作用。 尽管对多囊蛋白进行了多年的研究，但对其在细胞分裂中的功能知之甚少。我们的惊喜 观察将为多囊蛋白在发病机制中调节细胞增殖提供新的见解 ADPKD 的。我们将在我们的研究中使用创新的显微镜方法。我们建议大力推动 本科生研究造福广大大学第一代大学生 托莱多并培训他们沉浸在定量细胞生物学研究中。