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The Biology and Biochemistry of ADAR RNA Editing Enzymes

ADAR RNA 编辑酶的生物学和生物化学

基本信息

批准号：
8182034
负责人：
Brenda L. Bass
金额：
$ 22.43万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-04-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8182034
关键词：
Active Sites Address Adenosine Affect Amino Acids Animals Behavioral Biochemical Biochemistry Bioinformatics Biological Assay Biological Process Biology Caenorhabditis elegans Catalysis Catalytic Domain Chemotaxis Codon Nucleotides Complementary DNA Cytidine Deaminase DRADA2b protein Data Deamination Defect Disease Double-Stranded RNA Enzymes Exhibits Family Funding Future Gene Silencing Pathway Genes Goals Health Human Inosine Lead Link Mammals Mediating Messenger RNA MicroRNAs Molecular Mutation Outcome Study Pathway interactions Phenotype Phytic Acid Play Property Protein Isoforms Proteins Protocols documentation RNA Editing RNA Precursors RNA Processing RNA Sequences Reagent Role Site Small RNA Structure Testing Therapeutic Studies Time Untranslated RNA Validation Variant Viral abstracting base cofactor dsRNA adenosine deaminase genome-wide insight interest malignant neurologic neoplasms mutant nervous system disorder research study viral RNA

项目摘要

DESCRIPTION (provided by applicant): Project Summary/Abstract ADARs are RNA editing enzymes that convert adenosine to inosine in cellular and viral double- stranded RNA (dsRNA). One function of ADARs is to target codons in mRNA to allow multiple protein isoforms from the information in a single gene. Codon editing is found in many neuronally important mRNAs, and aberrant levels of editing have been linked to neurological disease. Fundamental to determining how ADARs contribute to human health, and sometimes disease, is a complete understanding of their catalytic mechanism. Experiments are proposed to determine the amino acids in the human ADAR2 catalytic domain that control its catalytic efficiency and lead to targeting of precise adenosines. A screen has identified mutations that affect these proceses, and these wil be characterized using a variety of biochemical assays. Despite its importance, editing in codons is rare, and there are far more inosines in noncoding RNA sequences. Yet, the function of inosines in noncoding sequences is unclear. Since ADARs target any dsRNA sequence, one posibility is that they affect dsRNA-mediated gene silencing pathways. High- throughput sequencing will be performed to compare the small RNAs of wildtype C. elegans with those in strains lacking ADARs. The focus will be on small RNAs that are processed from a dsRNA precursor, namely, microRNAs and endogenous siRNAs. Altered levels of small RNAs as well as their editing sites will be tabulated, and the latter will be distinguished from sequencing errors by their absence in animals lacking ADAR editing. Effects of ADARs on small RNAs will be correlated with predicted changes in mRNA levels using microarray analyses. While C. elegans lacking ADARs are viable, they have chemotaxis defects, and it is anticipated they have other subtle defects that have not been recognized. After validation of observations made with bioinformatics studies, phenotypes suggested by molecular defects will be tested. PUBLIC HEALTH RELEVANCE: Project Narrative ADARs are RNA editing enzymes that are essential for human life. They have been linked to longevity, and aberrant levels of editing have been associated with neurological disease and cancer. The proposed work will enhance our understanding of how ADARs contribute to human health, as well as disease, by advancing our understanding of the enzyme's catalytic mechanism and its substrates.

描述（由申请人提供）： ADARs是一种RNA编辑酶，可以将细胞和病毒双链RNA（dsRNA）中的腺苷转化为肌苷。ADAR的一个功能是靶向mRNA中的密码子，以允许来自单个基因中的信息的多种蛋白质同种型。密码子编辑存在于许多神经重要的mRNA中，并且异常水平的编辑与神经系统疾病有关。要确定ADAR如何促进人类健康，有时甚至是疾病，关键是要完全了解其催化机制。提出实验来确定人ADAR 2催化结构域中控制其催化效率并导致精确腺苷靶向的氨基酸。筛选已经鉴定出影响这些过程的突变，并且这些突变将使用各种生化测定来表征。尽管其重要性，但密码子中的编辑是罕见的，并且在非编码RNA序列中有更多的肌苷。然而，肌苷在非编码序列中的功能尚不清楚。由于ADAR靶向任何dsRNA序列，一种可能性是它们影响dsRNA介导的基因沉默途径。将进行高通量测序以比较野生型C的小RNA。elegans与缺乏ADAR的菌株中的那些。重点将放在从dsRNA前体加工的小RNA上，即microRNA和内源性siRNA。将改变的小RNA水平及其编辑位点制表，后者将通过在缺乏阿达尔编辑的动物中不存在而与测序错误区分开来。ADAR对小RNA的影响将与使用微阵列分析预测的mRNA水平变化相关。而C.缺乏ADAR的秀丽线虫是可行的，它们具有趋化性缺陷，并且预计它们具有尚未被认识到的其他细微缺陷。在生物信息学研究验证观察结果后，将检测分子缺陷提示的表型。公共卫生关系： ADAR是RNA编辑酶，对人类生命至关重要。它们与长寿有关，异常的编辑水平与神经系统疾病和癌症有关。拟议的工作将通过推进我们对酶催化机制及其底物的理解，增强我们对ADAR如何促进人类健康以及疾病的理解。