Phase separation control of transcription in Ewing sarcoma

尤文肉瘤转录的相分离控制

• 批准号: 10541896
• 负责人: Jacob C. Schwartz
• 金额: $ 33.27万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-16 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10541896
• 关键词: Adolescent; Affect; Affinity; Binding; Biochemical; Bone neoplasms; Cells; Childhood; Chimeric Proteins; Chromatin; Complex; Cytoplasmic Granules; DNA; DNA Methylation; DNA Repair; DNA Structure; Development; Diagnosis; Disease; EWS-FLI1 fusion protein; EWSR1 gene; Effectiveness; Enhancers; Enzymes; Equilibrium; Event; Ewings sarcoma; FLI1 gene; Family; Gene Expression; Gene Mutation; Gene Silencing; Genes; Genetic; Genetic Heterogeneity; Genetic Transcription; Genome; Genomics; Goals; Homologous Gene; In Vitro; Invaded; Knowledge; Link; Liquid substance; Localized Disease; Malignant Bone Neoplasm; Micrometastasis; Modeling; Molecular; Molecular Biology; Neurodegenerative Disorders; Nucleic Acid Binding; Nucleic Acids; Oncogenes; Organelles; Outcome; Pathology; Patients; Persons; Phase; Physical condensation; Play; Process; Property; Protein Engineering; Protein Family; Proteins; Proteomics; RNA; RNA Binding; RNA Polymerase II; RNA-Binding Protein EWS; RNA-Binding Protein FUS; RNA-Binding Proteins; Repressor Proteins; Research; Role; Small Interfering RNA; Solid; Structure; TAF15 gene; Techniques; Tertiary Protein Structure; Titrations; Transcriptional Activation; Transcriptional Regulation; Work; biochemical model; childhood sarcoma; chromatin remodeling; granule cell; histone modification; monomer; next generation sequencing; novel; novel strategies; nucleic acid structure; overexpression; primary bone cancer; prognostic of survival; programs; protein oligomer; recruit; sarcoma; scaffold; small molecule; transcription factor; tumor; tumorigenesis

Phase separation control of transcription in Ewing sarcoma Eighty-five percent of Ewing sarcomas are caused by translocations between the genes EWS and FLI1, creating a powerful oncogene – EWS-FLI1. The remainder are caused by translocations involving an EWS homologue, FUS. This sarcoma is an aggressive primary bone tumor affecting 1 in 300,000 people annually with more than 80% of tumors occurring in adolescents, making it the second most common pediatric bone cancer. Half of known translocations identified in sarcomas involve one of a family of three proteins – FUS, EWS, and TAF15, known as the FET family of proteins. Our work over the last few years has contributed to expanding the model for FET protein regulation of transcription. FET proteins control transcription in an organizational capacity. FET proteins undergo an assembly process called phase separation, which is the process thought to drive formation of non-membrane bound organelles, also called granules. In this proposal, we highlight our accomplishments isolating an EWS-FLI1 associated granule from cells. These granules have considerable similarity to those we have discovered to associate with RNA polymerase II (RNA Pol II) during transcription, which we refer to as transcription granules. This proposal aims to establish a model that EWS- FLI1 fusion protein seed aberrant granules that incorporate EWSR1 and other FET proteins to alter transcription and promote tumorigenesis. Our proposed work will investigate basic molecular mechanisms by which EWS-FLI1 can contribute and drive pediatric sarcoma pathology. Our proposal involves three aims. (1) We will determine the binding partners that comprise fusion protein granules for EWS-FLI1. (2) We will investigate EWS-FLI1 and EWSR1 activities to bind nucleic acids, phase separate, and alter transcription through effects on nucleic structures. Finally, (3) we will investigate how changes to DNA structure and chromatin are linked to aberrant phase separation by EWS- FLI1, EWSR1, and additional remodeling factors.

尤文肉瘤转录的相分离控制 百分之八十五的尤文肉瘤是由肉瘤之间的易位引起的 基因 EWS 和 FLI1，产生强大的癌基因 – EWS-FLI1。 由涉及 EWS 同源物 FUS 的易位引起。 侵袭性原发性骨肿瘤每年影响 30 万分之一的人，发病率超过 80% 青少年发生的肿瘤，使其成为第二常见的儿科骨骼 肉瘤中已知的易位有一半涉及一个三口之家中的一个。 蛋白质 – FUS、EWS 和 TAF15，称为 FET 蛋白质家族。 过去几年致力于扩大 FET 蛋白调控模型 FET 蛋白控制组织能力中的转录。 经历称为相分离的组装过程，该过程被认为是 驱动非膜结合细胞器（也称为颗粒）的形成。 在本提案中，我们强调了我们在隔离 EWS-FLI1 相关的成就 这些颗粒与我们所拥有的颗粒非常相似。 发现在转录过程中与 RNA 聚合酶 II (RNA Pol II) 相关， 我们称之为转录颗粒。该提案旨在建立一个 EWS- 的模型。 FLI1 融合蛋白种子异常颗粒，包含 EWSR1 和其他 FET 我们提出的工作将改变转录并促进肿瘤发生。 研究 EWS-FLI1 贡献和驱动的基本分子机制 我们的建议涉及三个目标（1）我们将确定。 (2) 我们将 研究 EWS-FLI1 和 EWSR1 结合核酸、相分离和 最后，（3）我们将研究如何通过影响核酸结构来改变转录。 DNA 结构和染色质的变化与 EWS 的异常相分离有关 FLI1、EWSR1 和其他重塑因子。