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

RUI：研究非肌肉肌球蛋白 II 收缩性的分子机制

• 批准号: 1716964
• 负责人: Derek Applewhite
• 金额: $ 58.94万
• 依托单位: Reed College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1716964&HistoricalAwards=false
• 关键词: RUI; Investigating; Molecular; Mechanisms; Non

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.

细胞改变形状的能力，这一过程称为形态发生，是生命的基本原理。 形态发生在发育过程中特别重要，是生成最终组成多细胞生物体的各个组织层所必需的。细胞利用收缩系统改变形状，该收缩系统由称为细胞骨架的细丝网络和拉动这些细丝（称为非肌肉肌球蛋白 II）的分子马达组成。 这项研究解决了这种收缩系统何时何地被激活以及哪些其他潜在蛋白质可以调节这种活动。 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，并结合计算方法，以发现可能在调节细胞骨架、非肌肉细胞中发挥作用的新蛋白质。肌球蛋白 II，或两者兼而有之。 这项研究将揭示形态发生这一关键过程是如何受到调控的，同时还将产生可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，他们将在高度协作的环境中接受指导，从而培养批判性思维和体验式学习。 学生将接受广泛的多学科培训，这将有助于他们为各种与科学相关的职业道路做好准备。还将为西北地区的研究人员举办定量生物学研讨会。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。它对于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特性众所周知，但决定其激活时间和地点的分子线索却鲜为人知。还缺乏参与调节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新型 NMII 调节分子 RN-tre 来剖析 NMII 的招募和激活。以 RN-tre 为例，该项目将在心尖收缩（一个关键的形态发生过程）期间确定 NMII 收缩性的其他调节因子。果蝇组织培养细胞将利用其对 RNAi 消耗的敏感性和有限的几何形状，用于建立体外顶端收缩测定，使其成为全内反射显微镜 (TIRF) 等高分辨率成像技术的理想选择。该项目还将开发一个计算框架，用于优先考虑候选 NMII 调节剂，从而能够使用顶端收缩测定来测试预测的蛋白质。 除了进一步了解关键发育过程的调控方式之外，该项目还将产生有价值的计算工具，帮助回答其他复杂的生物学问题。 这些研究的结果将扩展我们对调节 NMII 收缩性机制的理解，从而增进细胞和发育生物学领域的知识。

项目成果

The Drosophila melanogaster Rab GAP RN-tre cross-talks with the Rho1 signaling pathway to regulate nonmuscle myosin II localization and function

果蝇 Rab GAP RN-tre 与 Rho1 信号通路相互作用调节非肌肉肌球蛋白 II 定位和功能

• DOI: 10.1091/mbc.e20-03-0181
• 发表时间: 2020-10-01
• 期刊: Molecular Biology of the Cell
• 影响因子: 3.3
• 作者: Platenkamp A;Detmar E;Sepulveda L;Ritz A;Rogers SL;Applewhite DA
• 通讯作者: Applewhite DA

Network-based prediction of polygenic disease genes involved in cell motility

基于网络的细胞运动相关多基因疾病基因预测

• DOI: 10.1186/s12859-019-2834-1
• 发表时间: 2019-06
• 期刊: BMC Bioinformatics
• 影响因子: 3
• 作者: Bern, Miriam;King, Alexander;Applewhite, Derek A.;Ritz, Anna
• 通讯作者: Ritz, Anna

Graphery : interactive tutorials for biological network algorithms

Graphery：生物网络算法的交互式教程

• DOI: 10.1093/nar/gkab420
• 发表时间: 2021-05
• 期刊: Nucleic Acids Research
• 影响因子: 14.9
• 作者: Zeng, Heyuan;Zhang, Jinbiao;Preising, Gabriel A;Rubel, Tobias;Singh, Pramesh;Ritz, Anna
• 通讯作者: Ritz, Anna

The Drosophila protein, Nausicaa, regulates lamellipodial actin dynamics in a Cortactin-dependent manner

果蝇蛋白 Nausicaa 以 Cortactin 依赖性方式调节板状肌动蛋白动力学

• DOI: 10.1242/bio.038232
• 发表时间: 2019-01
• 期刊: Biology Open
• 影响因子: 2.4
• 作者: O'Connell, Meghan E.;Sridharan, Divya;Driscoll, Tristan;Krishnamurthy, Ipsita;Perry, Wick G.;Applewhite, Derek A.
• 通讯作者: Applewhite, Derek A.

From network analysis to experimental validation: identification of regulators of non-muscle myosin II contractility using the folded-gastrulation signaling pathway

从网络分析到实验验证：使用折叠原肠胚形成信号通路识别非肌肉肌球蛋白 II 收缩力的调节因子

• DOI: 10.1186/s12860-023-00492-3
• 发表时间: 2023-10-11
• 期刊: BMC Molecular and Cell Biology
• 影响因子: 2.8
• 作者: Zhao, Andy;Varady, Sophia;O’Kelley;Deng, Vicki;Platenkamp, Amy;Wijngaard, Petra;Bern, Miriam;Gormley, Wyatt;Kushkowski, Elaine;Thompson, Kat;et al
• 通讯作者: et al