Actin Assembly and Cell Motility: Mechanisms and Regulation

 描述(由申请人提供)：我们的实验室研究肌动蛋白组装和基于肌动蛋白的细胞运动的分子和细胞机制。我们想要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组件和膜相互作用，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)合并现有的赠款，2)促进新研究方向的灵活性，3)促进稳定性和指导。1)现有拨款。“跨内皮细胞迁移的机制”是目前一项长期关注肌动蛋白组装和膜的拨款的标题。对我们来说，跨内皮细胞迁移是一个新的领域，它是在对NK细胞如何迁移、发现和杀死其靶细胞的研究中出现的。我们感兴趣的是免疫细胞和癌细胞如何在生理和病理过程中穿过血管内皮细胞。迁移细胞和内皮细胞都积极参与，改变形状和施加力，这是肌动蛋白细丝与质膜和细胞内膜细胞器相互作用的结果。“肌动蛋白封顶蛋白的调控”是目前一项较新的基金的标题，该基金专注于肌动蛋白封顶蛋白调控的新机制。封端蛋白是肌动蛋白-细丝带刺末端可获得性和活性的关键调节因子。新近发现的新型调节剂 Capping Protein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，调节器将封顶蛋白定位于细胞需要组装肌动蛋白细丝的位置，并将其微丝封顶活性调整到与反应物浓度和反应动力学有关的生理水平。2) 灵活性和新的研究方向。正如我的生物素描中所描述的那样，我已经成功地寻求新的方向，使用新的实验系统，并适应新的技术。在这项提议中，我们的实验室的新领域包括细胞骨架细丝系统之间的新连接，使用斑马鱼作为脊椎动物模型系统，光诱导的蛋白质活性控制，以及超分辨率相关光学和电子显微镜。3)辅导与稳定。Mira的常见问题解答提到“有更多的时间进行研究，并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。我喜欢担任导师，我为我们的实验室实习生和我的教职员工在我们系内外取得的成功感到非常自豪。在稳定性方面，虽然我们感谢我们成功地为我们的研究争取到持续的资金，但通过Mira机制，我们进行过渡和探索新方向的能力将更加便捷和高效。

 DESCRIPTION (provided by applicant): 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.