Spectrin as a mechanosensitive hub integrating polarity, growth and trafficking

血影蛋白作为整合极性、生长和运输的机械敏感中心

• 批准号: 1952922
• 负责人: Claire Thomas
• 金额: $ 86.27万
• 依托单位: Pennsylvania State Univ University Park
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1952922&HistoricalAwards=false
• 关键词: Spectrin; mechanosensitive; hub; integrating; polarity

This project will increase our understanding of how cells coordinate their structure and their growth. Cells, which are the fundamental unit of life, have a defined shape built by a set of internal structural proteins known as the cytoskeleton. Cells also grow, divide, and sense their mechanical environment. Some of these functions are also dependent on cytoskeletal proteins. In this project, the researches will examine the mechanism by which spectrin, a giant cytoskeletal protein, performs its functions in cell growth, division and mechano-sensing. Associated with this research, the principle investigator and her team will engage in outreach activities that promote STEM careers including leading an effort in the College of Science to bring a mobile science classroom to central/eastern Pennsylvania and working with a local science museum. These activities will provide high-impact STEM experiences to K-12 students, as well as bring teacher professional development to rural and inner-city areas. The principle investigator’s lab will also participate in programs that promote women, minority and LGBTQ+ undergraduate participation in research, and bring research projects into undergraduate teaching laboratories, enhancing undergraduate education. Cellular growth must be integrated with the mechanical properties of cellular structures. Spectrins are giant cytoskeletal proteins with two contrasting activities. They are best known for their ability to form fishing net-like structural networks with F actin at the cell cortex that can undergo mechanical stretching. However, spectrins have recently also been shown to modulate protein trafficking in the endosome system. The molecular regulation that balances these two activities is unknown. Through the study of Beta-H-spectrin in epithelial cells of Drosophila melanogaster, this project will test two competing hypotheses to elucidate the molecular mechanisms by which spectrins coordinate mechanosensing and growth control in the cell. One approach will test the hypothesis that tension in the spectrin network plays a role in the relocation of the membrane protein Crumbs and in its control of the growth-inhibiting Hippo pathway. The second approach tests the hypothesis that spectrin networking contributes to the regulation of Crumbs/Hippo control of cell growth in endosomes. The project will use advanced STORM imaging and protein biochemistry to determine the geometry of the spectrin network and the extent to which it is stretched by changes in Myosin II-generated tension. Protein biochemistry, genetics and cell biology will also be used to further understand the role of spectrin and its associated proteins in the regulation of endosomal protein trafficking. Overall the results of this project will generate new paradigms for how spectrins function in the control of cellular growth.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这个项目将增加我们对细胞如何协调其结构和生长的理解。细胞是生命的基本单位，其形状是由一组被称为细胞骨架的内部结构蛋白构成的。细胞也可以生长、分裂和感知机械环境。其中一些功能也依赖于细胞骨架蛋白。在这个项目中，研究人员将研究巨型细胞骨架蛋白血影蛋白在细胞生长、分裂和机械传感中的作用机制。与这项研究相关联，首席调查员和她的团队将参与促进STEM职业生涯的外联活动，包括在科学学院领导一项努力，将流动科学教室带到宾夕法尼亚州中部/东部，并与当地的一家科学博物馆合作。这些活动将为K-12学生提供高影响力的STEM体验，并将教师专业发展带到农村和市中心地区。首席调查员实验室还将参与促进女性、少数民族和LGBTQ+本科生参与研究的项目，并将研究项目带入本科教学实验室，加强本科教育。细胞生长必须与细胞结构的机械特性相结合。幽灵蛋白是一种巨大的细胞骨架蛋白，具有两种截然不同的活性。它们最为人所知的是它们有能力与细胞皮质上的F肌动蛋白形成渔网状的结构网络结构，可以进行机械拉伸。然而，最近也有研究表明，幽灵蛋白可以调节内体系统中的蛋白质运输。平衡这两种活性的分子调控尚不清楚。通过对果蝇上皮细胞中β-H-光谱蛋白的研究，本项目将检验两种相互竞争的假说，以阐明其在细胞中协调机械传感和生长控制的分子机制。一种方法将检验这一假设，即血影蛋白网络中的张力在膜蛋白Crumbs的重新定位及其对生长抑制河马途径的控制中发挥作用。第二种方法验证了这样一种假设，即血影蛋白网络有助于调节Crumbs/Hippo对内体细胞生长的控制。该项目将使用先进的风暴成像和蛋白质生物化学来确定血影蛋白网络的几何形状，以及由于肌球蛋白II产生的张力的变化而拉伸的程度。还将利用蛋白质生物化学、遗传学和细胞生物学来进一步了解血影蛋白及其相关蛋白在调节内体蛋白运输中的作用。总体而言，该项目的成果将为幽灵蛋白如何在细胞生长控制中发挥作用产生新的范例。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。