C. elegans RNA regulatory protein complexes that control germ cell fate

控制生殖细胞命运的线虫 RNA 调节蛋白复合物

• 批准号: 8636913
• 负责人: Scott T Aoki
• 金额: $ 5.85万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8636913
• 关键词: Adaptor Signaling Protein; Address; Adult; Affect; Binding; Biochemical; Biochemistry; Caenorhabditis elegans; Cell Maintenance; Cells; Cellular biology; Complex; Development; Foundations; Future; Genetic; Germ; Germ Cells; Germ Lines; Goals; Homologous Gene; Human; Imaging Techniques; In Vitro; Length; Location; Measures; Messenger RNA; Microscopy; Modeling; Molecular; Organism; Poly A; Poly(A) Tail; Polynucleotide Adenylyltransferase; Post-Transcriptional Regulation; Process; Proliferating; Protein Family; Proteins; RNA; Recombinants; Regulation; Relative (related person); Stem cells; Techniques; Testing; Tissues; Transcript; Translations; Work; genetic regulatory protein; in vivo; member; novel; protein complex; protein protein interaction; stem cell fate; tumor progression

DESCRIPTION (provided by applicant): The long-term goal of my proposal is to understand how RNA-protein complexes regulate translation to influence cell fate. The C. elegans adult germ line provides a simple model to study questions involving the molecular controls influencing a germ stem cell's (GSC's) decision to proliferate or differentiate into gametes. The GSC fate decision is coordinated by FBF-1 and FBF-2 (collectively referred to as FBF), redundant members of the highly conserved PUF (Pumilio and FBF) protein family. FBF binds to specific messenger RNAs (mRNAs) necessary for differentiation, and is shown in vitro to associate with deadenylating (CCF-1/Not deadenylation complex) and polyadenylating (GLD-3/GLD-2 cytoplasmic poly(A) polymerase) proteins. In a working model, FBF interacts with specific poly(A) tail modifying proteins in different regions of the germ line to regulate expression of target mRNAs involved in GSC fate decisions. The following proposal combines established techniques in genetics, cell biology, and biochemistry, with newer developments in fluorescent microscopy, to further our understanding of the assembly and spatial regulation of FBF complexes. I will test our model with three specific aims: 1) I will determine the specific protein combinations that can assemble with FBF in a minimal in vitro protein interaction network. 2) Using advanced fluorescent imaging techniques, I will visualize where in the germ line and what subcellular location FBF interacts with its binding partners. 3) I will test in vivo whether FBF adenylation-modifying protein complexes affect the poly(A) tail length of FBF- associated transcripts. Together, my proposal will increase our general understanding of how RNA regulatory protein complexes control tissue development in a multi-cellular organism.

描述（由申请人提供）：我的提案的长期目标是了解RNA-蛋白质复合物如何调节翻译以影响细胞命运。梭elegans成体生殖系提供了一个简单的模型来研究涉及影响生殖干细胞（GSC）增殖或分化为配子的决定的分子控制的问题。GSC命运决定由高度保守的PUF（Pumilio和FBF）蛋白家族的冗余成员FBF-1和FBF-2（统称为FBF）协调。FBF与分化所必需的特异性信使RNA（mRNA）结合，并且在体外显示与去腺苷酸化（CCF-1/非去腺苷酸化复合物）和聚腺苷酸化（GLD-3/GLD-2细胞质聚（A）聚合酶）蛋白相关。在一个工作模型中，FBF与生殖系不同区域的特异性poly（A）尾修饰蛋白相互作用，以调节参与GSC命运决定的靶mRNA的表达。以下建议结合了遗传学，细胞生物学和生物化学中的既定技术，以及荧光显微镜的最新发展，以进一步了解FBF复合物的组装和空间调控。我将测试我们的模型有三个具体的目标：1）我将确定特定的蛋白质组合，可以组装FBF在一个最小的体外蛋白质相互作用网络。2)使用先进的荧光成像技术，我将可视化在生殖细胞系和亚细胞位置FBF与其结合伙伴相互作用。3)我将在体内测试FBF腺苷酸化修饰蛋白复合物是否影响FBF相关转录物的poly（A）尾长度。总之，我的建议将增加我们对RNA调节蛋白复合物如何控制多细胞生物体中组织发育的一般理解。