Processing and Function of Polyoma RNA

多瘤病毒 RNA 的加工和功能

• 批准号: 8320881
• 负责人: Gordon G Carmichael
• 金额: $ 31.93万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8320881
• 关键词: ADAR1; Active Sites; Address; Affect; Antisense RNA; Attention; Binding; Binding Proteins; Biological Models; Cell Nucleus; Cells; Collection; DNA; DNA Sequence; DNA biosynthesis; Double-Stranded RNA; Elements; Funding; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Goals; Heterodimerization; Infection; Inosine; Lead; Learning; Life Cycle Stages; Mammalian Cell; Methods; Molecular; Mus; Nature; Nuclear; Pathway interactions; Peptide Signal Sequences; Plasmids; Play; Polyadenylation; Polyomavirus; Process; RNA; RNA Editing; RNA Polymerase II; RNA Processing; Regulation; Reporter; Role; Signal Transduction; Site; System; Time; Transcript; Untranslated RNA; Viral; Viral Gene Expression Regulation; Viral Genes; Viral Genome; Virus; Work; design; dsRNA adenosine deaminase; men; novel; novel strategies; promoter; research study; response; small molecule; vector; viral RNA

DESCRIPTION (provided by applicant): The overall goals of this project are to understand how polyoma virus RNAs are made and regulated in infected cells and to use this virus as a model system to learn more general rules for mammalian RNA processing and function. We have discovered that much regulation of viral gene expression depends on the formation of double-stranded RNAs and their editing by the cellular dsRNA specific adenosine deaminase (ADAR1). Experiments completed during the previous funding period suggested that the long-sought trigger for the viral early-late switch might be the overlap and perhaps also the editing of the early and late polyadenylation signals. This represents a new mechanism of gene regulation. We also learned much new about nuclear responses to dsRNAs, including the involvement of a long nuclear-retained noncoding RNA (NEAT1) in the assembly of nuclear bodies called paraspeckles, which function in the retention of edited RNAs. We will extend these studies in three related aims. In the first, we will use new methods to better characterize RNA expression in the viral life cycle and will determine the nature and interplay of DNA sequence elements that promote regulated gene expression. In the second aim we will examine more closely the fate of polyoma dsRNAs in the nucleus, and the impact of cellular dsRNA response pathways including paraspeckles on infection. In the final aim we will carry out studies to learn more about how antisense RNA works in mammalian nuclei. This work will involve studying whether single DNA molecules can allow concurrent transcription in both directions as well as a new approach using small molecule-directed heterodimerization to alter the intranuclear proximity of DNA and RNA molecules.

描述（由申请人提供）：该项目的总体目标是了解如何在感染细胞中制造和调节多瘤病毒RNA，并将该病毒用作模型系统，以了解有关哺乳动物RNA处理和功能的更多通用规则。我们发现，病毒基因表达的大部分调节取决于双链RNA的形成及其通过细胞dsRNA特异性腺苷脱氨酶（ADAR1）的编辑。在上一个资金期间完成的实验表明，病毒早期开关的长期触发因素可能是重叠，也可能是早期和晚期聚腺苷酸化信号的编辑。这代表了基因调节的新机制。我们还了解了对DSRNA的核反应的新知识，包括将长核保留的非编码RNA（Neat1）参与到称为羊角面包的核体组装中，这在保留编辑的RNA中起作用。我们将把这些研究扩展到三个相关目标。首先，我们将使用新方法在病毒生命周期中更好地表征RNA表达，并确定促进受调节基因表达的DNA序列元素的性质和相互作用。在第二个目标中，我们将更仔细地检查细胞核中多瘤DSRNA的命运，以及包括副羊皮胶囊对感染的细胞DSRNA反应途径的影响。在最终目标中，我们将进行研究，以了解有关反义RNA如何在哺乳动物核中起作用的更多信息。这项工作将涉及研究单个DNA分子是否可以在这两个方向上同时进行转录，以及使用小分子定向的异二聚化的新方法，以改变DNA和RNA分子的核内接近度。