RUI: Collaborative Research: Intracellular Patterning in the Ciliate Cell Cortex

本项目旨在确定使用单细胞原生动物嗜热四膜虫在细胞表面组织专门结构的通用机制。 研究的结构包括对摄食和排泄都很重要的结构。 将产生关于新合成的细胞表面组分如何以适当的丰度递送到适当的位置的知识。 该项目将通过独立和基于课程的研究经验相结合，吸引早期职业科学家（主要是本科生）参与遗传学，细胞生物学和先进显微镜。 这一努力试点计划招募来自代表性不足背景的学生进入指导，合作研究团队，长期承诺学徒。 沿着的方式，我们将开发“可见的细胞”，一个全面的教学和研究资源，包括荧光细胞探针在工程细胞系中表达，并提供给更广泛的研究界。 该集合将包括表达荧光标记蛋白质的细胞，突出特定的细胞器，膜系统和细胞骨架成分，可以在实验室和教室中进行研究。这项研究的重点是加宽的皮质结构域（BCD）1基因，当禁用时，会导致多个嘴和多余的排泄孔的组装。 该基因的序列分析表明，其产物类似于参与膜运输的蛋白质。 实验的目的是测试的假设，Bcd1蛋白促进内吞作用（检索多余的表面材料），通过本地化的激酶激活。 这些实验包括膜运输和细胞信号传导途径的药理学和遗传学操作，以阐明这些途径的稳态如何导致细胞表面细胞器的正确组装。 最后，我们将确定和特征的基因，导致janusA表型，一个镜像复制的整个腹细胞器模式。 总之，这些研究的BCD 1和janusA将提供的机制，确定双方的本地化和尺寸的主要细胞表面结构和全球细胞patterning.This奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响审查标准的支持。

This project seeks to identify universal mechanisms that organize specialized structures on the cell surface using the single-celled protozoan Tetrahymena thermophila. Structures studied include ones important for both feeding and excretion. Knowledge will be generated as to how newly synthesized cell-surface components are delivered to the proper location and in the proper abundance. This project will engage early career scientists (primarily undergraduates) in genetics, cell biology, and advanced microscopy through a combination of independent and course-based research experiences. This effort pilots a program to recruit students from under-represented backgrounds into mentored, collaborative research teams with long-term committed apprenticeships. Along the way, we will develop the “Visible Cell”, a comprehensive teaching and research resource consisting of fluorescent cellular probes expressed in engineered cell-lines and made available to the broader research community. This collection will include cells that express fluorescently-tagged proteins spotlighting specific organelles, membrane systems, and cytoskeletal components which can be studied in both the laboratory and the classroom. This research focuses on the Broadened Cortical Domain (BCD)1 gene, which when disabled, results in the assembly of multiple mouths and supernumerary excretory pores. Sequence analysis of this gene reveals that its product resembles a protein involved in membrane trafficking. Experiments are designed to test the hypothesis that the Bcd1 protein promotes endocytosis (retrieval of excess surface materials) through localized kinase activation. These experiments include pharmacological and genetic manipulation of membrane trafficking and cell signaling pathways to elucidate how homeostasis of these pathways leads to proper assembly of cell surface organelles. Finally, we will identify and characterize the gene that causes the janusA phenotype, a mirror-image duplication of the entire ventral organelle pattern. Together, these studies of BCD1 and janusA will provide understanding of mechanisms that determine both the localization and dimensions of major cell surface structures and global cell patterning.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.