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

RUI：合作研究：纤毛细胞皮层的细胞内模式

• 批准号: 1947608
• 负责人: Eric Cole
• 金额: $ 41.74万
• 依托单位: Saint Olaf College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1947608&HistoricalAwards=false
• 关键词: RUI; Collaborative; Research; Intracellular; Patterning

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.

该项目旨在利用单细胞原生动物嗜热四膜虫来确定在细胞表面组织特殊结构的通用机制。所研究的结构包括对摄食和排泄都很重要的结构，并将产生有关如何传递新合成的细胞表面成分的知识。该项目将通过结合独立和基于课程的研究经验，吸引早期职业科学家（主要是本科生）从事遗传学、细胞生物学和高级显微镜学方面的研究。计划招募来自代表性不足的背景的学生加入受指导的合作研究团队，并进行长期的学徒训练。在此过程中，我们将开发“可见细胞”，这是一种由工程细胞中表达的荧光细胞探针组成的综合教学和研究资源。该系列将包括表达荧光标记蛋白的细胞，突出显示可以在实验室和课堂上研究的特定细胞器、膜系统和细胞骨架成分。扩展皮质结构域 (BCD)1 基因，当该基因被禁用时，会导致多个嘴和多余排泄孔的组装。对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。实验旨在验证以下假设： Bcd1 蛋白通过局部激酶激活促进内吞作用（回收多余的表面物质）。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操作，以阐明体内平衡的机制。最后，我们将鉴定并表征导致 janusA 表型的基因，即整个腹侧细胞器模式的镜像复制，这些对 BCD1 和 janusA 的研究将提供理解。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Anterior–posterior pattern formation in ciliates

纤毛虫的前后图案形成

• DOI: 10.1111/jeu.12890
• 发表时间: 2022-09
• 期刊: Journal of Eukaryotic Microbiology
• 影响因子: 2.2
• 作者: Cole, Eric;Gaertig, Jacek
• 通讯作者: Gaertig, Jacek

The Tetrahymena bcd1 mutant implicates endosome trafficking in ciliate, cortical pattern formation

四膜虫 bcd1 突变体涉及纤毛虫、皮质模式形成中的内体运输

• DOI: 10.1091/mbc.e22-11-0501
• 发表时间: 2023-07-01
• 期刊: Molecular Biology of the Cell
• 影响因子: 3.3
• 作者: Cole, Eric S.;Maier, Wolfgang;Joachimiak, Ewa;Jiang, Yu-Yang;Lee, Chinkyu;Collet, Erik;Chmelik, Carl;Romero, Daniel P.;Chalker, Douglas;Alli, Nurudeen K.;Ruedlin, Tina M.;Ozzello, Courtney;Gaertig, Jacek
• 通讯作者: Gaertig, Jacek