RUI: Investigating the Molecular Mechanisms of Non-muscle Myosin II Contractility

细胞改变形状的能力，即所谓的形态发生，是生命的基本原理。形态发生在发育过程中特别重要，是产生最终组成多细胞生物体的不同组织层所必需的。细胞通过收缩系统改变形状，收缩系统由称为细胞骨架的细丝网络和拉动这些细丝的分子马达组成，称为非肌肉肌球蛋白II。这项研究旨在研究这种收缩系统何时何地被激活，以及哪些其他潜在的蛋白质可能调节这种活动。为了回答这些问题，研究小组将使用基于细胞的系统来研究发育过程中发生的形态发生，结合计算方法，将允许发现新的蛋白质，这些蛋白质可能在调节细胞骨架、非肌肉肌球蛋白II或两者中发挥作用。这项研究将揭示这一关键过程--形态发生--是如何受到调控的，同时还将产生可以广泛应用于社区其他研究问题的计算工具。更广泛的影响活动包括组建一个主要由本科生组成的研究小组，他们将在高度协作的环境中接受指导，以培养批判性思维和体验式学习。这些学生将接受广泛的、多学科的培训，这将有助于他们为各种与科学相关的职业道路做好准备。还将为西北大学的研究人员举办数量生物学研讨会，以确定非肌肉肌球蛋白II(NMII)产生的收缩是细胞迁移和分裂过程中的一个基本细胞过程。它对形态发生或发育过程中发生的细胞形状变化特别重要。虽然NMII的许多动力学和生物物理性质是众所周知的，但决定其何时何地被激活的分子线索还远未被很好地理解。同样缺乏的是参与调节细丝动态和收缩的分子的完整清单。本项目的目的是通过分析一种新的NMII调控分子RN-tre来剖析NMII的招募和激活。以RN-TrE为例，该项目将确定NMII在根尖收缩过程中的其他调节因子，这是一个关键的形态形成过程。利用果蝇组织培养细胞对RNAi缺失的敏感性和有限的几何形状，将其用于建立体外根尖收缩实验，使其成为全内反射显微镜(TIRF)等高分辨率成像技术的理想材料。该项目还将开发一个计算框架，用于确定候选NMII调节因子的优先顺序，从而能够使用心尖收缩试验测试预测的蛋白质。除了进一步了解关键的发育过程是如何调控的，该项目还将产生有价值的计算工具，有助于回答其他复杂的生物学问题。这些研究的结果将扩大我们对NMII收缩调节机制的理解，从而促进细胞和发育生物学领域的知识。

The ability of cells to change shape, a process known as morphogenesis, is a fundamental principle of life. Morphogenesis is particularly important during development, and is needed to generate the various layers of tissues that eventually comprise multi-cellular organisms. Cells change shape using a contractile system composed of a network of filaments known as the cytoskeleton, and a molecular motor that pulls on these filaments known as non-muscle myosin II. This research addresses when and where this contractile system is activated and what other potential proteins may regulate this activity. To answer these questions, the research team will use a cell-based system to study the morphogenesis that occurs during development, coupled with computational approaches that will allow for the discovery of new proteins that may play a role in regulating the cytoskeleton, non-muscle myosin II, or both. This research will uncover how this crucial process, morphogenesis, is regulated while also producing computational tools that can be broadly applied to other research questions in the community. The Broader Impact activities includes the formation of a research team composed primarily of undergraduate students who will be mentored in a highly collaborative environment that will foster critical thinking and experiential learning. The students will receive a broad, multi-disciplinary training which will help to prepare them for a variety of science-affiliated career paths. A workshop on Quantitative Biology will also be held for researchers in the NorthwestContractility generated by Non-muscle Myosin II (NMII) is a fundamental cellular process that occurs during cell migration and division. It is particularly important to morphogenesis, or the cell shape change that occurs during development. While many of the kinetic and biophysical properties of NMII are well known, the molecular cues dictating when and where it is activated are far less well understood. What is also lacking is a complete list of the molecules involved in the regulation filament dynamics and contractility. The objective of this project is to dissect the recruitment and activation of NMII through the analysis of a novel NMII regulatory molecule RN-tre. With RN-tre as an example, the project will identify additional regulators of NMII contractility during apical constriction, a critical morphogenic process. Drosophila tissue culture cells will be used to establish an in vitro apical constriction assay by taking advantage their sensitivity to RNAi depletion and confined geometry, making them ideal for high-resolution imaging techniques such as total internal reflection microscopy (TIRF). This project will also develop a computational framework for prioritizing candidate NMII regulators, enabling the testing of the predicted proteins using the apical constriction assay. Along with furthering the understanding of how the critical developmental process is regulated, this project will also produce valuable computational tools that can help answer other complex biological questions. The results of these studies will advance knowledge in the fields of cell and developmental biology by extending our understanding of the mechanisms that regulate NMII contractility.