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

细胞改变形状的能力，这一过程被称为形态发生，是生命的基本原理。形态发生在发育过程中尤为重要，它是产生最终构成多细胞生物的各种组织层所必需的。细胞通过一种可收缩的系统来改变形状，该系统由一种被称为细胞骨架的纤维网络和一种拉动这些纤维的分子马达（被称为非肌肉肌球蛋白II）组成。这项研究解决了这个收缩系统何时何地被激活，以及其他哪些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，再加上计算方法，将允许发现可能在调节细胞骨架，非肌肉肌球蛋白II或两者中发挥作用的新蛋白质。这项研究将揭示这个关键的过程，形态发生，是如何调节的，同时也产生了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团队，他们将在一个高度协作的环境中接受指导，这将促进批判性思维和体验式学习。学生们将接受广泛的、多学科的培训，这将有助于他们为各种科学相关的职业道路做好准备。由非肌肉肌球蛋白II （NMII）产生的收缩性是细胞迁移和分裂过程中发生的基本细胞过程。它对发育过程中发生的形态发生或细胞形状变化尤为重要。虽然NMII的许多动力学和生物物理特性是众所周知的，但决定它何时何地被激活的分子线索却知之甚少。同样缺乏的是参与调控丝动力学和收缩性的分子的完整列表。该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的募集和激活。以RN-tre为例，该项目将确定顶端收缩过程中NMII收缩性的其他调节因子，这是一个关键的形态形成过程。果蝇组织培养细胞将利用其对RNAi耗尽的敏感性和受限的几何形状，使其成为高分辨率成像技术（如全内反射显微镜（TIRF））的理想选择，用于建立体外顶端收缩试验。该项目还将开发一个计算框架，用于确定候选NMII调节因子的优先级，从而能够使用根尖收缩试验测试预测的蛋白质。随着对关键发育过程如何调控的进一步理解，这个项目也将产生有价值的计算工具，可以帮助回答其他复杂的生物学问题。这些研究的结果将通过扩展我们对调节NMII收缩性的机制的理解，推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.