Mechanisms of Developmentally Regulated Splicing

发育调控剪接机制

• 批准号: 7024726
• 负责人: Thomas A Cooper
• 金额: $ 29.25万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7024726
• 关键词: RNA splicing; animal genetic material tag; binding sites; bioinformatics; biological signal transduction; cell differentiation; cell growth regulation; cell line; chick embryo; developmental genetics; gene expression; genetic screening; genetically modified animals; laboratory mouse; microarray technology; myocardium; polymerase chain reaction; posttranslational modifications; protein binding; protein isoforms; protein localization; protein metabolism; striated muscles; tissue /cell culture

DESCRIPTION (provided by applicant): The long term goal of this proposal is to understand the mechanisms by which endogenous alternative splicing regulators modulate natural splicing transitions. Many proteins have been shown to bind to specific pre-mRNAs and regulate splicing in experiments using exogenous proteins and RNA, however, very little is known about how natural splicing transitions are regulated in vivo. Striated muscle development provides an ideal system to discover networks of coordinately regulated alternative splicing transitions, identify the determinative regulators of specific sets of splicing transitions, determine how the activities of endogenous splicing factors are modulated to drive these transitions, and identify the signaling pathways that control the activities of the determinative splicing regulators. We will perform a systematic large scale screen to identify alternative splicing events that are robustly regulated during differentiation of skeletal muscle in cell culture and during fetal to adult development of heart and skeletal muscle tissues. We will establish correlations between splicing transitions and the expression of a panel of known splicing regulators. Comparisons of mouse and chicken striated muscle development will be used to identify conserved transitions of splicing patterns and regulator expression. Bioinformatic analyses will be used to identify putative factor binding sites as well as novel motifs among regulated pre-mRNAs. Sets of robustly regulated alternative splicing transitions will be used to identify splicing factors that are determinative for specific splicing transitions and coordinated regulation. Cause:effect relationships between splicing transitions and regulator expression will be tested, in cell culture and in transgenic and knockout mouse models. We will also determine the mechanisms by which the activities of splicing regulators are modulated during development. Understanding developmentally regulated splicing is crucial for understanding the mechanisms of gene expression in normal and pathological states and will ultimately lead to approaches to correct or circumvent disease processes at the molecular level.

描述（由申请人提供）：该提案的长期目标是了解内源选择性剪接调节剂调节自然剪接转变的机制。许多蛋白质已被证明可以与特定的前 mRNA 结合并在使用外源蛋白质和 RNA 的实验中调节剪接，然而，人们对体内自然剪接转变如何调节知之甚少。横纹肌发育提供了一个理想的系统来发现协调调节的选择性剪接转变网络，识别特定剪接转变组的决定性调节因子，确定如何调节内源剪接因子的活性以驱动这些转变，并识别控制的信号通路决定性剪接调节因子的活动。我们将进行系统的大规模筛选，以确定在细胞培养中骨骼肌分化期间以及心脏和骨骼肌组织从胎儿到成人发育期间受到强有力调节的选择性剪接事件。我们将建立剪接转变和一组已知剪接调节因子的表达之间的相关性。小鼠和鸡横纹肌发育的比较将用于识别剪接模式和调节器表达的保守转变。生物信息学分析将用于识别假定的因子结合位点以及受调节的前 mRNA 中的新基序。一组受到严格调控的选择性剪接转变将用于识别对特定剪接转变和协调调控起决定性作用的剪接因子。将在细胞培养物以及转基因和敲除小鼠模型中测试剪接转变和调节剂表达之间的因果关系。我们还将确定在开发过程中调节剪接调节剂活性的机制。了解发育调节剪接对于理解正常和病理状态下基因表达的机制至关重要，并将最终导致在分子水平上纠正或规避疾病过程的方法。