mRNA regulatory functions of the Drosophila TRIM-NHL protein, Brat

果蝇 TRIM-NHL 蛋白 Brat 的 mRNA 调节功能

• 批准号: 10418852
• 负责人: Aaron Charles Goldstrohm
• 金额: $ 29.44万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10418852
• 关键词: Adenosine; Adult; Affect; Animals; BRAT gene; Binding; Binding Proteins; Binding Sites; Bioinformatics; Biological; Biological Assay; Biological Process; Brain; Brain Neoplasms; Cancer Etiology; Cell Culture Techniques; Cells; Complex; Congenital Hydrocephalus; Cultured Cells; Data; Development; Developmental Process; Disease; Drosophila genus; Drosophila melanogaster; Embryonic Development; Family; Functional disorder; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Gene Expression Regulation; Genetic; Genetic Translation; Goals; Infertility; Lobular Neoplasia; Location; Malignant Neoplasms; Maps; Measures; Mediating; Messenger RNA; Modeling; Molecular; Mutate; Names; Oogenesis; Pathogenesis; Pathway interactions; Poly(A) Tail; Post-Transcriptional Regulation; Proteins; RNA; RNA Binding; RNA-Binding Proteins; Reporter; Reporter Genes; Repression; Research; Ribonucleases; Role; TRIM Gene; TRIM Motif; Tail; Tissues; Translations; Work; fly; gene function; gene network; genetic corepressor; genome editing; improved; in vivo; insight; loss of function; mRNA Decay; mRNA Expression; mRNA Stability; mRNA Transcript Degradation; member; mutant; nervous system disorder; neurogenesis; protein function; stem cell fate; stem cell proliferation; stem cells; stemness; transcription factor; transcriptome; translatome

Project Summary/Abstract: TRIM-NHL proteins (named after their TRIpartite Motif and founding members NCL-1, HT2A, LIN-41) are essential for animal development and control of stem cell fate. Their dysfunction causes cancer, infertility, and neurological disorders including congenital hydrocephalus. The recent discovery that multiple TRIM-NHL proteins bind to mRNAs provided a crucial insight into their molecular functions. The long term goal of this research is to discover how TRIM-NHL proteins regulate gene expression to control crucial developmental processes. This proposal focuses on the Drosophila melanogaster TRIM-NHL protein, Brain Tumor (Brat), which binds to specific mRNAs and functions in neurogenesis, oogenesis, and embryogenesis. In stem cells, Brat suppresses stemness and promotes differentiation whereas loss-of-function leads to over-proliferation in the brain and germline. We pursue the central hypothesis that Brat negatively regulates gene expression by causing mRNA degradation and inhibiting translation to control stem cell fate. Our research plan will determine the effect of Brat on translation, mRNA decay, and stem cell fate during neurogenesis and oogenesis. A major strength of our work is that it integrates both molecular regulatory mechanisms of Brat and its biological functions at the organismal level. First, using quantitative assays that specifically measure Brat activity, we will dissect its repressive domains and identify the required pathways and corepressors in cultured cells. Second, we will identify the network of genes that are regulated by Brat. We will map Brat-binding sites across the transcriptome and measure Bratʹs effect on the abundance and translation status of expressed mRNAs. By integrating the resulting data, our data will provide a comprehensive view of the relationship between Brat-RNA occupancy, location of functional binding sites, and impact on mRNA stability and translation efficiency. Third, we use precision genome editing to interrogate the roles of RNA-binding and repression domains of the brat gene during neurogenesis and oogenesis in Drosophila. We also created tissue specific reporter gene assays that specifically measure Bratʹs repressive activity in stem cells. The results of this research will provide a mechanistic understanding of Brat mediated gene regulation and provide a global view of its impact on gene expression. Our results will establish the role of Brat’s mRNA regulatory activities in the control of the stem cell proliferation-differentiation axis. Brat serves as an archetype for the TRIM-NHL family, and our discoveries will also broadly enhance the understanding of TRIM-NHL protein functions in development and disease.

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