ACTIN ASSEMBLY AND CELL MOTILITY: MECHANISMS AND REGULATION

研究计划 家长赠款摘要/摘要(R35 GM118171) 我们的实验室研究肌动蛋白组装和基于肌动蛋白的细胞的分子和细胞机制。 能动性。我们想了解肌动蛋白细丝是如何组装的，它们如何与其他细胞骨架相互作用。 成分和膜，以及这些相互作用如何决定细胞的形状和运动。 MIRA的目标是：1)结合现有的拨款，2)促进新研究的灵活性 方向，以及3)促进稳定和指导。 1)现有拨款。“跨内皮细胞迁移的机制”是目前一项长期的研究课题。 常备基金专注于肌动蛋白组装和膜。跨内皮细胞迁移是一个新的领域 美国是在研究NK细胞如何迁移、发现和杀死它们的目标细胞后出现的。我们是 感兴趣的是免疫细胞和癌细胞如何在生理过程中穿过血管内皮细胞 和病理过程。迁移细胞和内皮细胞都积极参与，改变 肌动蛋白细丝与质膜相互作用的形状和作用力 胞内膜性细胞器。 “肌动蛋白封顶蛋白的调控”是目前一项新的、专注于小说的拨款的标题。 肌动蛋白封端蛋白的调控机制。封顶蛋白是影响可利用性的关键因素 肌动蛋白-细丝带刺末端的活性。最近发现的新型封端蛋白调节剂已经 揭示了意想不到的见解，极大地改变了我们对细胞内肌动蛋白组装的看法。我们现在知道 调节器的目标是将蛋白质封顶到细胞需要组装肌动蛋白细丝的位置，而它们 将其灯丝封顶活性调整到与 反应物和反应动力学。 2)灵活性和新的研究方向。我已经成功地追求了新的方向， 使用新的实验系统，适应新技术，就像我的生物素描中所描述的那样。新的 在这项建议中，我们实验室的领域包括细胞骨架细丝系统之间的新连接， 利用斑马鱼作为脊椎动物模型系统，光诱导控制蛋白质活性，以及超 分辨相关光学显微镜和电子显微镜。 3)辅导与稳定。Mira常见问题解答提到，“有更多的时间进行研究和 在更稳定的环境中指导初级科学家“，这与我产生了强烈的共鸣。我喜欢 作为一名导师，我为我们实验室实习生和我的教职员工的成功感到非常自豪， 在我们部门内外。在稳定方面，虽然我们对我们在 为我们的研究获得持续的资金，我们进行转型和探索新方向的能力 将通过Mira机制变得更加便捷和高效。 1

Research Plan Summary / Abstract from Parent Grant (R35 GM118171) Our lab studies molecular and cellular mechanisms for actin assembly and actin-based cell motility. We want to understand how actin filaments assemble, how they interact with other cytoskeletal components and membranes, and how these interactions dictate cell shape and movement. The MIRA goals are: 1) combining existing grants, 2) promoting flexibility in new research directions, and 3) promoting stability and mentoring. 1) Existing Grants. “Mechanisms for Transendothelial Migration” is the current title of a long- standing grant focused on actin assembly and membranes. Transendothelial migration is a new area for us, one that emerged from studies on how NK cells migrate, find and kill their target cells. We are interested in how immune and cancer cells cross the endothelium of the vasculature during physiological and pathological processes. The migrating cell and the endothelial cell both actively participate, changing shape and exerting force as a result of actin filaments interacting with the plasma membrane and intracellular membranous organelles. “Regulation of Actin Capping Protein” is the current title of a newer grant focused on novel mechanisms of regulation of actin capping protein. Capping protein is a key regulator of the availability and activity of actin-filament barbed ends. Recent discoveries of novel regulators of capping protein have revealed unexpected insights that dramatically change our view of actin assembly in cells. We now know that regulators target capping protein to sites where cells need to assemble actin filaments and that they tune its filament-capping activity to a level that is physiologically relevant for the concentrations of the reactants and kinetics of the reactions. 2) Flexibility and New Research Directions. I have been successful in pursuing new directions, using new experimental systems, and adapting to new technologies, as described in my Biosketch. New areas for our lab in this proposal include novel connections between cytoskeleton filament systems, the use of zebrafish as a vertebrate model system, light-induced control of protein activity, and super- resolution correlative light and electron microscopy. 3) Mentoring and Stability. The MIRA FAQ mentions “more time for conduct of research and mentoring junior scientists in a more stable environment,” which resonates strongly with me. I enjoy serving as a mentor, and I am very proud of the success of our lab trainees and my faculty colleagues, within and outside of our department. In terms of stability, while we are grateful for our success in securing continued funding for our research, our ability to make transitions and explore new directions will be made more facile and efficient by the MIRA mechanism. 1