MOLECULAR BIOLOGY OF DSRNA ADENOSINE DEAMINASE

描述（由申请人提供）：本申请的主要目标是了解作用于RNA的RNA编辑酶腺苷脱氨酶（ADARs）在神经系统中的功能。秀丽隐杆线虫将用于这些研究，缺乏所有ADARs的菌株已被分离出来并发现表现出行为缺陷。为了将ADARs在特定神经元中的表达与特定行为联系起来，ADARs将在突变株的特定神经元中表达，并监测表型拯救。其他研究将集中于确定相关的编辑rna。提出了微阵列分析和肌苷特异性切割策略。这些方法将识别mrna编码区和非编码区肌苷，但后者将是进一步研究的重点。将进行实验，以确定非编码序列中含有肌苷的mrna如何在体内代谢，以及在没有RNA编辑的情况下这种代谢如何变化。最近的实验表明，ADARs与RNA干扰途径交叉，可能的机制将被测试。理解行为的分子决定因素是分子生物学最后的前沿之一。这一应用为在行为、基因沉默和dsRNA在细胞中的一般作用方面的新发现提供了潜力。

DESCRIPTION (provided by applicant): The primary goal of this application is to understand how the RNA editing enzymes called Adenosine Deaminases that act on RNA (ADARs) function in the nervous system. Caenorhabditis elegans will be used for these studies, and strains lacking all ADARs have been isolated and found to exhibit behavioral defects. To correlate expression of ADARs in particular neurons with particular behaviors, ADARs will be expressed in specific neurons of mutant strains, and phenotypic rescue monitored. Other studies will focus on determining the relevant edited RNAs. Microarray analyses and inosine-specific cleavage strategies are proposed. These methods will identify inosines in coding as well as non-coding regions of mRNAs, but the latter will be the focus of further study. Experiments to determine how mRNAs with inosines in non-coding sequences are metabolized in vivo, and how this metabolism changes in the absence of RNA editing will be performed. Recent experiments indicate ADARs intersect with RNA interference pathways, and possible mechanisms for this will be tested. Understanding the molecular determinants of behavior is one of the last frontiers of molecular biology. This application offers the potential for new discoveries in behavior, gene silencing and the general roles of dsRNA in cells.