BBSRC-NSF/BIO Leveraging synthetic biology to probe the rules of cell morphogenesis

这个项目试图了解细胞如何使用合成生物学工具控制它们的形状和运动。细胞运动对于单个细胞的捕猎和交配以及多细胞生物体的正确发育和功能都是必不可少的。片状突起称为片状脂膜，是驱动和引导这种运动的引擎。然而，它们的形成规则并不是很清楚。就像蚂蚁群体中，没有个人负责，一个群体的整体行为取决于蚂蚁之间局部相互作用的简单规则，细胞生物学的许多方面都受到蛋白质之间相互作用的局部规则的支配。这个项目试图通过从合成的、设计的蛋白质构建板脂来定义这些蛋白质-蛋白质相互作用的规则，这些蛋白质的相互作用模式可以被定制。这项工作是细胞形状/运动专家(加州大学旧金山分校的Orion Weiner)和蛋白质设计专家(布里斯托尔大学的Dek Woolfson)合作完成的。将通过多项协同活动加强科技人员队伍的多样性。(1)改革研究生院招生程序，使其更具包容性；(2)为代表性不足群体的高中生提供研究机会，并为教师和小学/中学/高中生进行实践科学演示；(3)通过探索馆、科学节探索日和科学咖啡馆向公众介绍情况，让更多的受众参与；(4)让决策者、地方和国家行业倡导使用生物技术中设计的蛋白质和系统；(5)对3名博士后研究员和5名高中生进行跨学科培训。这项工作将加强美国和英国实验室之间以及NSF和BBSRC之间的联系。Weiner的实验室最近发现了一种关键的肌动蛋白核仁的纳米级组织，该组织为片状脂体的形成提供了一个自组织模板。这些模板没有被充分理解，无法直接操纵它们的生物物理参数(如直线度与曲率、刚性、动力学等)。充分探讨肌动蛋白核糖核酸寡聚与细胞形态和功能的关系。典型的遗传和生化方法的一个强有力的替代方法是从头开始建立肌动蛋白调节器，这样就可以系统地探索细胞形状和运动的分子逻辑。为了做到这一点，蛋白质工程和从头开始的蛋白质设计(Woolfson实验室)将被用来产生合成蛋白质，这些蛋白质在我们对自然系统的知识的指导下，在细胞膜上以定义的几何形状组装以形成肌动蛋白。这些合成系统将在体内使用有缺陷的片状脂体形成和最终完全重组的细胞来测试它们支持片状脂膜形成的能力。这些研究将推动蛋白质设计在细胞内的应用，反过来，它们将有助于定义细胞形状和运动的分子逻辑。这项美英合作项目得到了美国国家科学基金会和英国生物技术和生物科学研究委员会的支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project seeks to understand how cells control their shape and movement using synthetic biology tools. Cell movement is essential for single cells to hunt and mate and for the correct development and function of multicellular organisms. Sheet-like protrusions called lamellipodia are the engines that power and guide this motility. However, the rules of their formation are not understood. Similar to ant colonies where no individual is “in charge”, and the overall behaviors of a colony depends on simple rules of local interaction between ants, many aspects of cell biology are dominated by local rules of interaction between proteins. This project seeks to define these rules of protein-protein interactions by constructing lamellipodia from synthetic, designer proteins whose patterns of interaction can be built to order. This work is a collaboration between an expert in cell shape/movement (Orion Weiner at the University of California-San Francisco) and an expert in protein design (Dek Woolfson at the University of Bristol). STEM workforce diversity will be enhanced through multiple synergistic activities. (1) reforming the grad school admissions process to make it more inclusive; (2) research opportunities for high school students from underrepresented groups and hands-on science demonstrations for teachers and elementary/middle/high-school students; (3) engagement of a wider audience by presentations to the general public through the Exploratorium, Science Festival Discovery Days, and Science Cafés; (4) Engagement of policy makers and local and national industries to advocate for the use of designed proteins and systems in biotechnology; (5) interdisciplinary training for three postdoctoral fellows and five high school students. This work will strengthen links between US and UK labs and between the NSF and the BBSRC.Weiner’s lab recently discerned the nanoscale organization of a key actin nucleator that suggests a self-organizing template for lamellipodia formation. These templates are not sufficiently understood to manipulate directly their biophysical parameters (such as linearity vs. curvature, rigidity, dynamics, etc.) and to probe fully the relation between actin nucleator oligomerization and cell shape and function. A powerful alternative to the typical genetic and biochemical approaches would be to build actin regulators from scratch, so that the molecular logic of cell shape and movement can be probed systematically. To do this, protein engineering and de novo protein design (Woolfson lab) will be used to generate synthetic proteins that assemble with defined geometries at membranes to nucleate actin, guided by our knowledge of the native system. These synthetic systems will be tested for their ability to support lamellipodial formation in vivo using cells defective in lamellipodia formation and ultimately in complete reconstitutions. These studies will advance protein design towards in-cell application, and, in turn, they will help define the molecular logic of cell shape and movement.This collaborative US/UK project is supported by the US National Science Foundation and the UK Biotechnology and Biological Sciences Research Council.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.