Mechanisms of 5'UTR splicing of human surfactant protein (SP)-A1 and SP-A2

人表面活性蛋白(SP)-A1和SP-A2 5UTR剪接机制

基本信息

批准号：
8335678
负责人：
GUIRONG WANG
金额：
$ 19.39万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-01 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8335678
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Acute Address Adult Affect Age Alternative Splicing Binding Binding Proteins Biological Assay Biological Models Birth Bronchopulmonary Dysplasia Chronic lung disease DNA Elements Enhancers Exclusion Exons Experimental Models Fetal Lung Future Gene Expression Generations Genes Genetic Translation Genomics Goals Health Status Host Defense Human Immune Infant Inflammatory Introns Knowledge Lung Lung diseases Malignant Neoplasms Mass Spectrum Analysis Messenger RNA Methodology Mutate Mutation Pathogenesis Pattern Physiological Play Premature Birth Process Protein Binding Proteins Proteome Pulmonary Surfactant-Associated Protein A Pulmonary Surfactant-Associated Protein D Pulmonary Surfactant-Associated Proteins RNA Splicing Regulation Reporter Role Sulfur System Testing Trans-Activators Translations Untranslated Regions Variant Western Blotting activator 1 protein base cis acting element falls fetal genetic variant human disease insight lung development mRNA Precursor mRNA Stability novel public health relevance research study respiratory distress syndrome surfactant

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to investigate mechanisms of alternative mRNA splicing of human surfactant protein A (SP-A) and potential implications/effects in the pathogenesis of several pulmonary diseases. Alternative mRNA splicing is a major regulatory mechanism that can generate protein diversity and increase proteome complexity. More than 70% of human genes express multiple mRNA variants through alternative splicing. About 20% of the variability from alternative splicing falls within the untranslated regions (5'UTR and 3'UTR) that affects mRNA stability, translation efficiency, and mRNA localization. SP-A, an important innate immune protein, plays a critical role in the innate host defense and the inflammatory regulation of lung, in surfactant-related physiological functions, and in parturition. In humans, there are two functional genes, SP-A1 and SP-A2. Differential expression of SP-A1 and SP-A2 has been observed in fetal and adult lung, and SP-A1 and SP-A2 genetic variants have been associated with pulmonary diseases including respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), and other lung diseases. The 5'UTR of SP-A1 and SP-A2 generates a number of splice variants, and these 5'UTR variants differentially influence translation efficiency and mRNA stability. Moreover, the ratio of SP-A1 to total SP-A content has been shown to vary as a function of age and lung health status. In this proposal, we hypothesize that differences exist between SP-A1 and SP-A2 in 5'UTR cis-acting elements and in trans-acting factors binding these elements. The rationale is based on the observation that: a) the key difference between the most frequently found SP-A1 (A*D') and SP-A2 (A*BD, A*BD') splice variants is the exclusion and inclusion, respectively, of exon B; b) our preliminary findings have shown that SP-A1 cis-acting element #6, i.e. exon splicing enhancer/silencer (ESE/S # 6), plays a role in splicing, and that specific protein(s) may bind to ESE/S #6. To test this hypothesis, we have proposed two specific aims: Specific Aim #1: to examine the functional impact of candidate cis-acting ESE/S elements in the SP-A1 and SP-A2 5'UTR splicing. Specific Aim #2: to study trans-acting factors involved in the regulation of SP-A 5'UTR splicing, with a focus on binding proteins associated with the SP-A1-specific cis-acting element ESE/S # 6. Upon completion of this R21 project, a novel experimental model system for alternative splicing of human SP-A can be established, and aspects of candidate cis-acting ESE/S elements within each SP-A gene and of trans-acting factors be analyzed and identified. These could provide foundational methodologies and insight for our further studies (R01 application) of the mechanisms involved in the splicing regulation of human SP-A1 and SP-A2, and of better understanding of the pathogenesis of several lung diseases. PUBLIC HEALTH RELEVANCE: This project is to establish a novel high-throughout experimental system for the study of mechanisms of alternative mRNA splicing of human surfactant proteins (SP-A1 and SP-A2). The proposed experiments include examination of the functional impact of candidate cis-acting ESE/S elements and identification of trans-acting factors binding these elements. New methodologies used and knowledge gained will contribute to the growing field of mRNA splicing regulation and to future studies that can help us to better understand the pathogenesis of several lung diseases.

描述（由申请人提供）：该项目的长期目标是研究人类表面活性剂蛋白A（SP-A）替代mRNA剪接的机制，以及在几种肺部疾病的发病机理中的潜在影响/作用。替代mRNA剪接是一种主要的调节机制，可以产生蛋白质多样性并增加蛋白质组的复杂性。超过70％的人基因通过替代剪接表达了多种mRNA变异。替代剪接的可变性约20％落在影响mRNA稳定性，翻译效率和mRNA定位的未翻译区域（5'UTR和3'UTR）内。 SP-A是一种重要的先天免疫蛋白，在先天宿主的防御和肺部的炎症调节，与表面活性剂相关的生理功能和分娩中起着至关重要的作用。在人类中，有两个功能性基因SP-A1和SP-A2。在胎儿和成人肺中已经观察到SP-A1和SP-A2的差异表达，SP-A1和SP-A2遗传变异与包括呼吸窘迫综合征（RDS），支气管肺肺增生不良（BPD）以及其他肺部疾病以及其他肺部疾病有关。 SP-A1和SP-A2的5'UTR产生了许多剪接变体，这些5'UTR变体差异地影响了翻译效率和mRNA稳定性。此外，SP-A1与总SP-A含量的比率已显示为随着年龄和肺部健康状况的函数而变化。在此提案中，我们假设5'UTR顺式作用元件以及结合这些元素的跨性别因素中的Sp-A1和Sp-A2之间存在差异。理由是基于以下观察结果：a）最常见的SP-A1（A*D'）和SP-A2（A*BD，A*BD'）剪接变体之间的关键差异分别是外显子B的排除和包容性； B）我们的初步发现表明，SP-A1顺式作用元素＃6，即外显子剪接增强子/消音器（ESE/S＃6），在剪接中起作用，并且该特定蛋白质可能与ESE/S＃6结合。为了检验这一假设，我们提出了两个具体的目的：特定目的＃1：检查候选候选sp-a1和sp-a2 5'utr剪接中的候选cis作用ESE/s元素的功能影响。具体目的2：研究与SP-A 5'UTR剪接有关的跨性别因素，重点是与SP-A1特异性的顺式作用元件ESE/S＃6相关的结合蛋白质。在完成此R21项目后，在人类SP-A中建立了Spece and Spece and Spine nectiast and Secis cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-cis-scis-cos-scis-s-scis-s-scis-sex-con-scis--分析和鉴定跨作用因子的因素。这些可以为我们的进一步研究（R01应用）提供基础方法和见解，该研究（R01应用）对人类SP-A1和SP-A2的剪接调节的机制以及对几种肺部疾病的发病机理的更好理解。公共卫生相关性：该项目是建立一个新型的高知识实验系统，以研究人类表面活性剂蛋白（SP-A1和SP-A2）的替代mRNA剪接机制。拟议的实验包括检查候选cis作用ESE/S元素的功能影响以及鉴定这些元素结合的跨作用因子。所用的新方法和获得的知识将有助于mRNA剪接调节的不断增长和未来的研究，从而帮助我们更好地了解几种肺部疾病的发病机理。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

DIFFERENTIAL SUSCEPTIBILITY OF HUMAN SP-B GENETIC VARIANTS ON LUNG INJURY CAUSED BY BACTERIAL PNEUMONIA AND THE EFFECT OF A CHEMICALLY MODIFIED CURCUMIN.

人类 SP-B 基因变异体对细菌性肺炎引起的肺损伤的不同易感性以及化学修饰姜黄素的作用

DOI：
10.1097/shk.0000000000000535
发表时间：
2016-04
期刊：
Shock (Augusta, Ga.)
影响因子：
0
作者：
Xu Y;Ge L;Abdel-Razek O;Jain S;Liu Z;Hong Y;Nieman G;Johnson F;Golub LM;Cooney RN;Wang G
通讯作者：
Wang G

A Mouse Model for Ocular Surface Staphylococcus aureus Infection.