Characterizing modes of maternal RNA degradation during vertebrate development

脊椎动物发育过程中母体 RNA 降解模式的表征

• 批准号: 8607201
• 负责人: Miler T. S. Lee
• 金额: $ 5.7万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8607201
• 关键词: 3' Untranslated Regions; Amanitins; Animals; Binding; Biological Models; Categories; Cell physiology; Cells; Computing Methodologies; Data; Deposition; Development; Developmental Process; Disease; Drosophila genus; Elements; Embryo; Embryology; Embryonic Development; Fertilization; Gene Expression Regulation; Genes; Genetic Transcription; Genome; High-Throughput Nucleotide Sequencing; Homologous Gene; In Situ Hybridization; Instruction; Laboratories; Length; Life; Malignant Neoplasms; Maps; Maternal Messenger RNA; Mediating; Messenger RNA; MicroRNAs; Molecular; Mothers; Organism; Pathway interactions; Pattern; Peptide Signal Sequences; Phenotype; Play; Poly A; Polyadenylation; Process; Property; Protein Isoforms; Proteins; RNA; RNA Degradation; RNA chemical synthesis; Regulation; Regulatory Element; Reporter; Reporter Genes; Resolution; Role; Signal Transduction; Site; Specificity; Staging; Structure; Time; Transcript; Translations; Untranslated Regions; Vertebrates; Work; Xenopus; Zebrafish; base; developmental disease; egg; human DICER1 protein; in vivo; mRNA Stability; mutant; novel; programs; research study; zygote

DESCRIPTION (provided by applicant): Early development in all animals depends initially on the maternal contribution of RNAs and proteins, deposited by the mother into the egg. The genome of the zygote is not actively transcribed initially, and all cellular processes including cel division are mediated by these initial maternal factors. At a later stage, the zygotic genome is activated and control switches from maternally contributed factors to newly synthesized zygotic RNAs and proteins. A key step for subsequent development is the destruction of maternal components. While this process has been known for over three decades, the molecular elements required to clear maternal instructions during vertebrate development remain largely unknown. In this proposal, I will investigate the mechanisms of maternal RNA degradation in vertebrates, using zebrafish as a model system. In Aim 1, I will use high throughput sequencing to identify maternally derived transcripts with correlated degradation patterns. I will compare timecourse expression data in wildtype embryos as well as embryos in which de novo transcription and small regulatory RNA synthesis have been inhibited, in order to define transcripts undergoing degradation by specific mechanisms and under specific temporal control. In Aim 2, I will analyze the lengths and sequence content of the 3' untranslated regions (UTRs) of maternal transcripts and look for regulated differences between the 3'UTRs of zygotic transcripts, under the hypothesis that differential regulation and degradation is mediated by these sequences. In Aim 3, I will characterize the discrete regulatory sequence signals present in 3'UTRs that are the determinants of degradation, using a combination of computational methods and reporter gene constructs. I expect that maternal RNAs that are co-degraded should also contain the same sets of signals. Together these aims will elucidate the ways in which RNA transcripts are differentially degraded. Although these processes are especially prominent during development, they are relevant to all living cells, which need to maintain regulated quantities of RNA messages in order to achieve specific phenotypes. Thus, understanding the mechanisms underlying RNA degradation is key to understanding how this process is misregulated in developmental disorders, cancers, and disease.

描述(申请人提供)：所有动物的早期发育最初依赖于母体的RNA和蛋白质的贡献，由母体沉积到卵子中。受精卵的基因组最初并不活跃地转录，所有的细胞过程，包括细胞分裂，都是由这些最初的母体因子调节的。在后来的阶段，合子基因组被激活，控制从母体贡献的因子切换到新合成的合子RNA和蛋白质。随后发展的一个关键步骤是销毁产妇组成部分。虽然这一过程已经知道了30多年，但在脊椎动物的发育过程中，清除母体指令所需的分子元件仍然基本上是未知的。在这项提议中，我将以斑马鱼为模型系统，研究脊椎动物母体RNA降解的机制。在目标1中，我将使用高通量测序来识别具有相关降解模式的母源转录本。我将比较野生型胚胎以及从头转录和小调节性RNA合成受到抑制的胚胎的时序表达数据，以确定在特定机制和特定时间控制下发生降解的转录本。在目标2中，我将分析母体转录本的3‘非翻译区(UTRs)的长度和序列含量，并在假设差异调控和降解是由这些序列介导的假设下，寻找合子转录本的3’UTRs之间的调控差异。在目标3中，我将使用计算方法和报告基因构建的组合来表征存在于3‘UTRs中的离散调控序列信号，这些信号是降解的决定因素。我预计，被共同降解的母体RNA也应该包含相同的信号集。这些目标将共同阐明RNA转录本的不同降解方式。虽然这些过程在发育过程中特别突出，但它们与所有活细胞有关，这些细胞需要维持受调控的RNA信息量，以实现特定的表型。因此，了解RNA降解的机制是理解这一过程如何在发育障碍、癌症和疾病中受到错误调控的关键。