Regulation of alternative pre-mRNA processing by small nucleolar RNAs

小核仁 RNA 对替代前 mRNA 加工的调节

• 批准号: 7892501
• 负责人: Stefan Stamm
• 金额: $ 26.46万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7892501
• 关键词: Affinity; Alternative Splicing; Area; Binding; Biochemical; Bioinformatics; Biology; Boxing; Cell Culture Techniques; Code; Collaborations; Complex; DNA; Data; Defect; Diagnostic; Drug Delivery Systems; Exons; Failure; Fostering; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genome; Goals; Human; In Vitro; Individual; Inherited; Knockout Mice; Knowledge; Life; Link; Masks; Mass Spectrum Analysis; Messenger RNA; Methods; Mission; Molecular; Molecular Target; Obesity; Orphan; Outcome; Pattern; Peptide Nucleic Acids; Peptides; Prader-Willi Syndrome; Process; Property; Proteins; Public Health; Publishing; RNA; RNA Sequences; RNA Splicing; Regulation; Regulatory Element; Reporter Genes; Reporting; Research; Resources; Ribosomal RNA; Role; SNRPN; Science; Serotonin Receptor 5-HT2C; Site; Small Nuclear RNA; Small Nucleolar RNA; Small RNA; System; Techniques; Testing; Textbooks; Therapeutic; Transcript; Transfer RNA; Work; base; human disease; improved; in vivo; innovation; mRNA Precursor; novel; prevent; public health relevance; success

DESCRIPTION (provided by applicant): There is a fundamental gap in understanding the function of small RNAs that constitute a large part of human gene expression. For example, we do not know how the loss of small nucleolar RNA (snoRNA) expression contributes to the Prader-Willi syndrome, the most frequent genetic cause for life-threatening obesity. Our long-term goal is to elucidate the regulation of alternative splicing by small RNAs that are estimated to comprise more than 40% of human gene expression. The objective of this application is to determine which alternative splicing patterns are influenced by the snoRNAs missing in individuals with Prader-Willi syndrome and to understand the molecular basis by which one of these snoRNAs, HBII-52, influences alternative splicing patterns. The central hypothesis is that snoRNAs missing in individuals with Prader- Willi syndrome regulate alternative splicing by masking splicing regulatory elements on pre- mRNAs. Thus these snoRNAs regulate expression of numerous genes by changing their alternative splicing patterns. The rationale for the proposed research is that the genes regulated by these snoRNAs represent drug targets for treatment of the Prader-Willi syndrome. This proposal is therefore relevant to the NIH's mission that pertains to developing fundamental knowledge that will potentially help to reduce the burdens of human disease. Guided by our first published report that the snoRNA HBII-52 influences alternative splicing and additional strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Identify the pre-mRNAs that are regulated by the snoRNAs missing in people with Prader-Willi syndrome; and 2) Determine how HBII-52 regulates alternative splice site usage and test alternatives for its expression. Under the first aim, changes in pre-mRNA processing will be determined after ectopically expressing individual snoRNAs using bioinformatic predictions and splice-site sensitive DNA arrays. Under the second aim, we will identify the repressor activity that competes with the snoRNA HBII-52 and determine its mode of action, using an established in vitro system. The proposed work is innovative, because it shows a complete new role for snoRNAs in the regulation of alternative pre-mRNA splicing. It may well change the current `textbook knowledge' that snoRNAs only regulate non-mRNAs. The proposed research is significant because it would show a novel role of snoRNAs, identify the molecular defects in Prader-Willi syndrome and help to predict the influence of small RNAs on splice site selection. PUBLIC HEALTH RELEVANCE: The proposed studies delve into an under-investigated area of gene expression and have the potential applicability to understand the molecular cause of the Prader-Willi Syndrome and potentially other forms of inherited obesity. The proposed research is relevant for public health, because it investigates a new mechanism of human gene expression. Understanding this mechanism is the basis to improve diagnostics and therapeutic options for human diseases caused by defects in pre-mRNA processing.

描述（由申请人提供）：对于构成人类基因表达大部分的小RNA的功能的理解存在根本性的差距。例如，我们不知道小核仁 RNA (snoRNA) 表达的丧失如何导致普瑞德-威利综合征，这是危及生命的肥胖症最常见的遗传原因。我们的长期目标是阐明小RNA对选择性剪接的调节，这些小RNA估计占人类基因表达的40%以上。本应用的目的是确定哪些选择性剪接模式受到 Prader-Willi 综合征个体中缺失的 snoRNA 的影响，并了解这些 snoRNA 之一 HBII-52 影响选择性剪接模式的分子基础。核心假设是，Prader-Willi 综合征个体中缺失的 snoRNA 通过掩盖前 mRNA 上的剪接调节元件来调节选择性剪接。因此，这些 snoRNA 通过改变其选择性剪接模式来调节许多基因的表达。这项研究的基本原理是，这些 snoRNA 调控的基因代表了治疗普瑞德威利综合征的药物靶点。因此，该提案与美国国立卫生研究院的使命相关，该使命涉及发展可能有助于减轻人类疾病负担的基础知识。在我们第一份发表的关于 snoRNA HBII-52 影响选择性剪接的报告和其他强有力的初步数据的指导下，这一假设将通过追求两个具体目标进行检验：1) 识别 Prader 患者中缺失的 snoRNA 所调节的前 mRNA -威利综合症； 2) 确定 HBII-52 如何调节选择性剪接位点的使用并测试其表达的选择性。第一个目标是，使用生物信息学预测和剪接位点敏感 DNA 阵列异位表达单个 snoRNA 后，将确定前 mRNA 加工的变化。在第二个目标下，我们将使用已建立的体外系统来鉴定与 snoRNA HBII-52 竞争的阻遏活性，并确定其作用模式。这项工作具有创新性，因为它展示了 snoRNA 在调节选择性前 mRNA 剪接中的全新作用。它很可能会改变目前 snoRNA 只调节非 mRNA 的“教科书知识”。这项研究意义重大，因为它将展示 snoRNA 的新作用，识别 Prader-Willi 综合征的分子缺陷，并有助于预测小 RNA 对剪接位点选择的影响。 公共健康相关性：拟议的研究深入研究了基因表达的研究不足的领域，并具有潜在的适用性来了解普瑞德-威利综合症和其他潜在形式的遗传性肥胖的分子原因。拟议的研究与公共卫生相关，因为它研究了人类基因表达的新机制。了解这一机制是改善由前 mRNA 加工缺陷引起的人类疾病的诊断和治疗方案的基础。