Elucidating CIS-Acting Elements Regulating Alcohol-Induced Alternative Splicing

阐明调节酒精诱导的选择性剪接的 CIS 作用元件

基本信息

批准号：
7918181
负责人：
HOWARD J EDENBERG
金额：
$ 19.25万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
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：
2009-08-20 至 2012-07-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Alternative splicing is an important level of gene regulation and a major source of proteome diversity in humans and other mammals. At least 74% of human genes express multiple mRNAs through alternative splicing of exons or exon segments. Deregulation of the splicing process is associated with a variety of disease states and conditions, and over 60% of known disease- causing mutations disrupt splicing. Our preliminary data indicate that alcohol treatment of rat hepatoma cells causes changes in the expression of many RNA-binding proteins, including some with known splicing activity. Therefore, it is likely that alcohol or its metabolism alters splicing. We will test the hypothesis that distinct combinations of cis-acting elements dictate alcohol-induced patterns of alternative splicing. We will use exon arrays to examine the effects of ethanol on the global pattern of splicing in rat hepatoma cells that metabolize ethanol, and in livers of rats exposed to ethanol. Based upon these data, we will develop a computational model to identify cis-acting RNA elements that regulate this process. We will examine the effects of ethanol on splicing in human hepatoma cells to determine whether these cis-acting RNA elements are conserved. The outcome of this study will not only provide a description of how alcohol affects the global splicing pattern, but also provide a set of testable hypotheses about the cis-acting sequences responsive to ethanol. In this exploratory project, we will test our hypothesis by the following three specific aims: (1) Identify alcohol-induced alternative pre- mRNA splicing in rat hepatoma cells and rat liver; (2) Computational discovery of alcohol- induced cis-acting regulatory elements; and (3) Test the generalization of the model to human cells. PUBLIC HEALTH RELEVANCE: We will use a combination of experimental and computational approaches to investigate alternative splicing in liver cells as a result of alcohol treatment, and to reveal molecular mechanisms, particularly cis-acting elements, that underlie alcohol-induced alternative splicing, which might contribute to liver injury.

描述（由申请人提供）：替代剪接是基因调节的重要水平，也是人类和其他哺乳动物中蛋白质组多样性的主要来源。至少74％的人基因通过外显子或外显子段的替代剪接表达多个mRNA。剪接过程的放松管制与多种疾病状态和条件有关，超过60％的已知疾病 - 导致突变破坏了剪接。我们的初步数据表明，酒精治疗大鼠肝癌细胞会导致许多RNA结合蛋白的表达变化，包括一些具有已知剪接活性的。因此，酒精或代谢可能会改变剪接。我们将检验以下假设：顺式作用元件的不同组合决定了酒精引起的替代剪接模式。我们将使用外显子阵列来检查乙醇对代谢乙醇的大鼠肝癌细胞以及暴露于乙醇的大鼠肝脏的影响。基于这些数据，我们将开发一个计算模型，以识别调节此过程的顺式作用RNA元素。我们将检查乙醇对人肝癌细胞中剪接的影响，以确定这些顺式作用RNA元件是否保守。这项研究的结果不仅将描述酒精如何影响全球剪接模式，而且还提供了一组关于响应于乙醇的顺式作用序列的可测试假设。在这个探索性项目中，我们将通过以下三个特定目的来检验我们的假设：（1）确定酒精诱导的替代性mRNA剪接在大鼠肝癌细胞和大鼠肝脏中；（2）酗酒诱导的顺式作用调节元件的计算发现；（3）测试模型对人类细胞的概括。公共卫生相关性：我们将使用实验和计算方法的组合来研究肝脏细胞中的替代剪接，并揭示出饮酒引起的替代性剪接的基础的分子机制，尤其是顺式作用元素，这可能导致肝损伤。