RUI: Investigating the mechanisms regulating the formation of lysosome-related organelles

RUI：研究调节溶酶体相关细胞器形成的机制

• 批准号: 1613804
• 负责人: Greg Hermann
• 金额: $ 46.94万
• 依托单位: Lewis and Clark College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1613804&HistoricalAwards=false
• 关键词: RUI; Investigating; mechanisms; regulating; formation

The project will investigate how compartments are generated within some cell types but not others in multicellular animals. The cells of complex organisms contain compartments called organelles that endow cells with their specific functional properties. While many organelles are common to all of the cells within an organism, some are only generated in a subset of cell types. One of these, called a lysosome-related organelle (LRO), plays varied and important roles, however only certain animal cells generate them. The research will investigate how this is achieved and identify factors that promote the formation of LROs. Many undergraduates (25-55/year)will be mentored in collaborative, investigative, and original research as part of the project. Many of these students will come from underrepresented groups in the sciences. The work will provide hands on scientific training for a large and diverse group of students, most of who will progress on to careers in science and education.In many cell types, lysosome-related organelles LROs co-exist with conventional lysosomes due to the activity of conserved LRO-specific trafficking pathways that divert cargo away from endolysosomes. The emergence of LRO biogenesis pathways likely result from the poorly understood interplay of general endolysosome trafficking factors expressed in all cells with key regulators that are only present and active within LRO-containing cells. To identify and investigate the function of these regulators, the project is analyzing the biogenesis of C. elegans gut granules, cell type-specific LROs that co-exist with conventional lysosomes. Some LRO biogenesis factors identified in this system have cell type restricted expression and likely regulate the diversion of cargo to LROs. The research addresses the function of one of these, GLO-3, which is proposed to redirect the function of CCZ-1, a RAB-7 guanine nucleotide exchange factor (GEF), away from conventional endolysosomal pathways, toward the GLO-1 Rab to regulate LRO biogenesis. The research will involve detailed phenotypic analysis of mutants, yeast 2-hybrid screens, and in vitro GEF assays. The research will examine whether expressing GLO-3 and other LRO biogenesis factors are sufficient to direct the trafficking of LRO cargo away from conventional endosomes in cell types that do not normally generate LROs. The research will employ a novel genetic resource to identify nearly all of the genes necessary for LRO biogenesis in C. elegans.

该项目将研究在多细胞动物中，某些细胞类型如何产生隔室，而其他细胞类型则否。 复杂生物体的细胞包含称为细胞器的隔室，这些隔室赋予细胞特定的功能特性。 虽然许多细胞器是生物体内所有细胞所共有的，但有些细胞器仅在细胞类型的子集中产生。 其中之一，称为溶酶体相关细胞器（LRO），发挥着各种重要的作用，但只有某些动物细胞才能产生它们。 该研究将调查这是如何实现的，并确定促进LRO形成的因素。 作为该项目的一部分，许多本科生（25-55岁/年）将接受协作、调查和原创研究的指导。 这些学生中的许多人将来自科学领域代表性不足的群体。 这项工作将为一个庞大而多样化的学生群体提供实际的科学培训，其中大多数人将在科学和教育事业中取得进展。在许多细胞类型中，溶酶体相关的细胞器LRO与传统的溶酶体共存，这是由于保守的LRO特异性运输途径的活动将货物从内溶酶体转移。 LRO生物发生途径的出现可能是由于对所有细胞中表达的一般性内溶酶体运输因子与仅存在于含LRO细胞内并在含LRO细胞内具有活性的关键调节因子的相互作用知之甚少。 为了识别和研究这些调节剂的功能，该项目正在分析C.线虫肠颗粒，与常规溶酶体共存的细胞类型特异性LRO。 在该系统中鉴定的一些LRO生物合成因子具有细胞类型限制性表达，并且可能调节货物向LRO的转移。 该研究解决了其中之一GLO-3的功能，GLO-3被提议将CCZ-1（RAB-7鸟嘌呤核苷酸交换因子（GEF））的功能从传统的内溶酶体途径转向GLO-1 Rab以调节LRO生物合成。 这项研究将涉及突变体的详细表型分析，酵母双杂交筛选和体外GEF测定。 该研究将检查表达GLO-3和其他LRO生物发生因子是否足以指导LRO货物远离通常不产生LRO的细胞类型中的常规内体的运输。 这项研究将利用一种新的遗传资源来确定几乎所有的基因所必需的LRO生物发生在C。优雅的