Erk-mediated regulation of Dicer and Drosha function in C. elegans

Erk 介导的线虫 Dicer 和 Drosha 功能调节

基本信息

批准号：
8535274
负责人：
Swathi Arur
金额：
$ 28.66万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8535274
关键词：
Antibodies Area Biogenesis Biological Biological Models Caenorhabditis elegans Cell Nucleus Cell physiology Cellular biology Complex Cytoplasm Data Development Developmental Process Dicer Pathway Down-Regulation Embryo Embryonic Development Enzymes Event Extracellular Signal Regulated Kinases Fertilization Fibroblasts Generations Genetic Genetic screening method Genomics Goals Homebound Persons Human Individual Lead Link Malignant Neoplasms Mediating Mediator of activation protein Meiosis Methods MicroRNAs Mitogen-Activated Protein Kinases Modeling Molecular Mus Oncogenic Oocytes Oogenesis Pathway interactions Pattern Pfeiffer Syndrome Phosphorylation Phosphotransferases Production Proteins Proteomics Regulation Relative (related person)Research Role Signal Pathway Signal Transduction Small RNA Sterility Syndrome System Testing Tumor Suppressor Proteins Work base extracellular fly functional genomics human DICER1 protein in vivo interest neoplastic cell novel oocyte maturation programs protein complex trophoblast tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Oogenesis and embryogenesis require the precise orchestration of multiple signaling networks and molecular pathways. Perturbations in any one event can lead to a host of developmental abnormalities such as the Apert and Pfieffer syndromes and cancer. Among the crucial components are the RAS-extracellular regulated kinase (ERK) signaling pathway and Dicer and Drosha, essential mediators of small RNA biogenesis. They govern a wide array of cellular and developmental processes, including many critical aspects of oogenesis and embryogenesis. For example, in mice, loss of ERK signaling results in defective oocyte maturation and perturbed trophoblast development, while in Caenorhabditis elegans loss of ERK function disrupts oocyte differentiation and maturation, leading to sterility or early embryonic lethality. Similarly, in C. elegans and mice, loss of Dicer or Drosha disrupts oocyte meiotic maturation and embryonic development, respectively. However, the functional intersection between the RAS/ERK pathway and small RNA biogenesis remains unclear, and no studies have shown that the RAS/ERK pathway regulates the activity of Dicer or Drosha. Our work on the identification of substrates through which ERK governs germline development in Caenorhabditis elegans recently uncovered a direct, functional link between the RAS/ERK pathway and Dicer and Drosha. In a functional genomics screen, we identified Dicer and Drosha as novel, evolutionarily conserved substrates of ERK that each function in a reciprocally antagonistic manner with ERK. By generating antibodies specific to the phosphorylated form of Dicer, we also observed that ERK-mediated phosphorylation of Dicer induces it to translocate from the cytoplasm to the nucleus in the C. elegans germline; this phosphorylation event and its consequences on Dicer localization are conserved in mouse embryonic fibroblasts and human tumor cells. Therefore, the goals of our proposed study are three-fold: i) to elucidate the effect of ERK-dependent phosphorylation on the localization of Drosha; ii) to test the impact of ERK-mediated phosphorylation on Dicer and Drosha function and small RNA production; and, iii) to determine the mechanistic basis of phospho-Dicer function by identifying functional partners of the modified proteins. Given the general relevance of RAS/ERK signaling pathway and Dicer and Drosha, results from our work will likely inform multiple systems, including human oogenesis and oncogenesis. !

描述（由申请人提供）：卵子发生和胚胎发生需要多个信号网络和分子途径的精确编排。在任何一种事件中，扰动都会导致许多发育异常，例如APERT和PFIEFFER综合征和癌症。在关键组件中，有RAS-纤维细胞调节激酶（ERK）信号通路和DICER和DROSHA，这是小RNA生物发生的必要介体。它们控制着各种各样的细胞和发育过程，包括卵子发生和胚胎发生的许多关键方面。例如，在小鼠中，ERK信号的丧失导致卵母细胞的成熟和肌细胞发育扰动，而在Caenorhabditis timans中，秀丽隐杆线虫在ERK功能的损失破坏了卵母细胞的分化和成熟，导致不育或早期胚胎致死性。同样，在秀丽隐杆线虫和小鼠中，DICER或DROSHA的丧失分别破坏了卵母细胞的减数分裂成熟和胚胎发育。然而，RAS/ERK途径与小RNA生物发生之间的功能相交尚不清楚，并且没有研究表明RAS/ERK途径调节DICER或DROSHA的活性。我们在鉴定ERK在Caenorhabditis elegans中鉴定的底物的工作最近发现了RAS/ERK途径与DICER和DICER和DROSHA之间的直接功能联系。在功能性基因组学筛选中，我们将DICER和DROSHA确定为ERK的新型，进化保守的底物，每种ERK都以相互拮抗的方式与ERK函数。通过产生对迪切尔磷酸化形式的抗体，我们还观察到ERK介导的DICER的磷酸化诱导其从细胞质转移到秀丽隐杆线虫种系中的核。这种磷酸化事件及其对DICER定位的后果在小鼠胚胎成纤维细胞和人肿瘤细胞中保守。因此，我们拟议的研究的目标是三个方面：i）阐明ERK依赖性磷酸化对Drosha定位的影响； ii）测试ERK介导的磷酸化对DICER和DROSHA功能以及RNA产生小的影响；以及，iii）通过鉴定修饰蛋白的功能伙伴来确定磷酸含量功能的机械基础。鉴于RAS/ERK信号通路和DICER和DROSHA的一般相关性，我们工作的结果可能会为多种系统提供信息，包括人类的卵子发生和肿瘤发生。呢