Regulation of alternative pre-mRNA processing by small nucleolar RNAs

小核仁 RNA 对替代前 mRNA 加工的调节

基本信息

批准号：
8307823
负责人：
Stefan Stamm
金额：
$ 26.2万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8307823
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 Genetic Transcription 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 Nucleolar Ribonucleoproteins 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

项目摘要

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）：了解构成人类基因表达很大一部分的小RNA功能的基本差距。例如，我们不知道小核仁RNA（SNORNA）表达的丧失如何促进prader-willi综合征，这是危及生命的肥胖症的最常见遗传原因。我们的长期目标是阐明小型RNA对替代剪接的调节，估计占人类基因表达的40％以上。该应用的目的是确定哪些替代剪接模式受Prader-Willi综合征中缺少的snornas的影响，并了解这些snornas hbii-52之一会影响替代剪接模式的分子基础。中心假设是，prader-willi综合征中缺少的ssshornas通过掩盖剪接调节元件在Pre-mRNA上调节替代剪接。因此，这些snornas通过改变其替代剪接模式来调节众多基因的表达。拟议研究的理由是，这些snoRNA调节的基因代表了治疗prader-willi综合征的药物靶标。因此，该提议与NIH的使命有关，该使命与发展基本知识有关，这将有助于减轻人类疾病的负担。在我们首次发表的报告的指导下，SNORNA HBII-52影响替代剪接和其他强大的初步数据，该假设将通过追求两个具体的目的来检验：1）确定由Prader-Willi综合征患者中缺少的SNORNAS所调节的MRNA； 2）确定HBII-52如何调节替代剪接位点使用情况并测试替代方案的表达。在第一个目标下，将使用生物信息学预测和剪接位点敏感的DNA阵列在异位表达单个snornas后确定MRNA前处理的变化。在第二个目标下，我们将使用建立的体外系统确定与SNORNA HBII-52竞争的阻遏活动，并确定其作用方式。拟议的工作是创新的，因为它在调节替代性mRNA剪接的调节中显示出了全新的作用。它很可能会改变snornas仅调节非MRNA的当前“教科书知识”。拟议的研究之所以重要，是因为它将显示出snornas的新作用，确定prader-willi综合征中的分子缺陷，并有助于预测小型RNA对剪接位点选择的影响。公共卫生相关性：拟议的研究深入研究了基因表达的不足区域，并具有了解Prader-Willi综合征的分子原因以及潜在的其他形式的遗传性肥胖症的潜在适用性。拟议的研究与公共卫生有关，因为它研究了人类基因表达的新机制。了解这种机制是改善由MRNA处理中缺陷引起的人类疾病的诊断和治疗选择的基础。