Regulation and functional effects of localized RNAs

局部RNA的调控和功能效应

• 批准号: 9153954
• 负责人: Stavroula Mili
• 金额: $ 98.65万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9153954
• 关键词: Adenomatous Polyposis Coli; Affect; Amyotrophic Lateral Sclerosis; Architecture; Area; Biochemical; Biological Assay; Cell Culture System; Cells; Characteristics; Colorectal Cancer; Complex; Cytoplasm; Cytoplasmic Granules; Destinations; Disease; Environment; Event; Extracellular Matrix; Goals; Late-Onset Disorder; Lead; Link; Malignant Neoplasms; Mediating; Microscopy; Mutation; Neurodegenerative Disorders; Pathway interactions; Process; Production; Proteins; RNA; RNA-Binding Proteins; Regulation; Ribonucleoproteins; Site; System; Translating; Translations; Tumor Suppressor Proteins; base; migration; mutant; novel; tumor progression

A large number of RNAs are not diffusely distributed in the cytoplasm, but are actively transported to various subcellular sites. After reaching their final destinations, localized RNAs are translated, thus directing local protein production. While increasing numbers of localized RNAs are being identified, the functional importance of these events is not well understood. We are focusing on a localization pathway that we have identified, which targets a number of RNAs to the tips of cellular protrusions. We have found that an important component of this pathway is the tumor-suppressor protein Adenomatous Polyposis Coli (APC) whose mutation is the initiating event in the progression of the majority of colorectal cancers. APC associates with RNAs at cellular protrusions in ribonucleoprotein complexes, which we term APC-RNPs. An additional component of APC-RNPs is the RNA-binding protein Fus/TLS, a protein whose mutation has been linked to both cancer and Amyotrophic Lateral Sclerosis (ALS). We have recently shown that ALS-associated mutants of Fus mislocalize APC-RNPs in internal cytoplasmic granules and misdirect their translation. During the last year, we have made significant progress in understanding how APC-RNP localization is regulated, and in identifying cellular factors involved in this process. Specifically, we have found that characteristics of the extracellular matrix environment can affect APC-RNP localization. This effect appears to be mediated through modulation of the cytoskeletal architecture, and we have made inroads in identifying the cytoskeletal components involved. We have additionally found that pathogenic mutations in the Fus protein lead to APC-RNP mislocalization through similarly affecting cytoskeletal architecture and dynamics. Interestingly, this effect is dependent on aggregation of the mutant Fus protein, a transition that is linked to triggering of late-onset diseases associated with Fus mutations (such as ALS and FTLD). Finally, we have devised a strategy to induce APC-RNP mislocalization in cellular systems and are currently characterizing the contribution of APC-RNP localization in various migratory parameters in 2D and 3D migration systems.

大量RNA并未扩散地分布在细胞质中，而是积极运输到各种亚细胞位点。到达最终目的地后，将局部RNA翻译，从而指导局部蛋白质的产生。尽管正在识别出局部RNA数量的增加，但这些事件的功能重要性尚不清楚。我们专注于已确定的定位途径，该途径将许多RNA靶向细胞突起的尖端。我们发现，该途径的重要组成部分是肿瘤 -​​ 抑制蛋白腺瘤性息肉大肠杆菌（APC），其突变是大多数结直肠癌进展的起始事件。 APC与核糖核蛋白复合物中细胞突起的RNA相关联，我们称其为APC-RNP。 APC-RNP的另一个组成部分是RNA结合蛋白FUS/TLS，一种蛋白质的突变与癌症和肌萎缩性侧面硬化症（ALS）相关。我们最近表明，与ALS相关的FUS突变体错误地定位了内部细胞质颗粒中的APC-RNP，并误导了它们的翻译。在过去的一年中，我们在了解如何调节APC-RNP定位以及确定该过程中涉及的细胞因素方面取得了重大进展。具体而言，我们发现细胞外基质环境的特征会影响APC-RNP的定位。这种效果似乎是通过调节细胞骨架结构来介导的，我们在识别涉及的细胞骨架成分方面陷入了进度。我们还发现，FUS蛋白中的致病突变通过类似地影响细胞骨架结构和动力学而导致APC-RNP错误定位。有趣的是，这种作用取决于突变FUS蛋白的聚集，该过渡与触发与FUS突变相关的晚期疾病（例如ALS和FTLD）有关。最后，我们设计了一种诱导APC-RNP错误定位在细胞系统中的策略，目前正在表征APC-RNP定位在2D和3D迁移系统中各种迁移参数中的贡献。