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

该项目旨在了解细胞如何使用合成生物学工具控制其形状和运动。 细胞运动对于单细胞狩猎和交配以及多细胞生物体的正确发育和功能至关重要。 片状突起称为板状伪足是动力和引导这种运动的引擎。 然而，它们的形成规律并不清楚。 类似于没有个体“负责”的蚁群，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由蛋白质之间相互作用的局部规则主导。 该项目旨在通过从合成的设计蛋白质构建片状伪足来定义蛋白质-蛋白质相互作用的规则，这些蛋白质的相互作用模式可以按顺序构建。 这项工作是细胞形状/运动专家（加利福尼亚大学弗朗西斯科分校的Orion Weiner）和蛋白质设计专家（布里斯托大学的Dek Woolfson）之间的合作。STEM员工的多样性将通过多种协同活动得到加强。 (1)改革格拉德入学程序，使其更具包容性;（2）为来自代表性不足群体的高中生提供研究机会，并为教师和小学/初中/高中学生提供实践科学演示;（3）通过探索馆、科学节探索日和科学咖啡馆向公众进行演示，吸引更广泛的受众;（4）让决策者和地方及国家工业界参与，倡导在生物技术中使用设计好的蛋白质和系统;（5）对三名博士后研究员和五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肌动蛋白成核剂的纳米级组织，这表明片状伪足形成的自组织模板。这些模板没有被充分理解以直接操纵它们的生物物理参数（例如线性与曲率、刚性、动力学等）。深入探讨肌动蛋白成核剂寡聚化与细胞形态和功能的关系。除了典型的遗传学和生物化学方法之外，一个强有力的替代方法是从头开始构建肌动蛋白调节因子，这样就可以系统地探索细胞形状和运动的分子逻辑。为此，蛋白质工程和从头蛋白质设计（Woolfson实验室）将用于生成合成蛋白质，这些蛋白质在我们对天然系统的了解的指导下，在膜上以确定的几何形状组装成核肌动蛋白。这些合成系统将被测试其能力，以支持板状伪足形成在体内使用细胞缺陷的板状伪足形成，并最终在完全重建。这些研究将推进蛋白质设计向细胞内应用，反过来，他们将帮助定义细胞形状和运动的分子逻辑。英国项目由美国国家科学基金会和英国生物技术和生物科学研究理事会支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的评估被认为值得支持。影响审查标准。

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.