Antisense RNAs control retrotransposon copy number

反义 RNA 控制逆转录转座子拷贝数

• 批准号: 8184610
• 负责人: David J. Garfinkel
• 金额: $ 28.22万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8184610
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Address; Affect; Antisense RNA; Area; Behavior; Biochemical; Biochemical Genetics; Biological Assay; Biology; Biomedical Research; Cells; DNA Transposable Elements; Defect; Defense Mechanisms; Disease; Ectopic Expression; Evolution; Gene Expression; Genes; Genetic; Genetic Epistasis; Genetic Screening; Genetic Transcription; Genome; Goals; HIV; HIV-1; Health; Human; Human Genome; Induced Mutation; Integrase; Mediating; Messenger RNA; Modeling; Monitor; Mutation; Pathway interactions; Play; Process; Proteins; RNA; RNA Interference; RNA Sequences; RNA chemical synthesis; Relative (related person); Research; Resistance; Retrotransposition; Retrotransposon; Retroviridae; Retrovirology; Reverse Transcription; Role; Saccharomyces; Saccharomyces cerevisiae; Saccharomycetales; Structure; System; Transcript; Translations; Untranslated RNA; Virus; Virus-like particle; Work; base; combat; complement C2a; drug development; genetic analysis; human DICER1 protein; improved; insight; interest; mutant; novel

DESCRIPTION (provided by applicant): Antisense RNAs control retrotransposon copy number The retrovirus-like transposon Ty1 of Saccharomyces cerevisiae is an effective model for understanding the replication of HIV, the causative agent of AIDS, and is arguably the best understood retrotransposon. Retrotransposons replicate through an RNA intermediate and have colonized most eukaryotic genomes. Studying Ty1 has also provided valuable information on human genome dynamics and evolution, and the behavior of transposon-induced mutations. RNA interference (RNAi) is a widespread mechanism used to silence infectious viruses and transposable elements, and alter gene expression, where small sequence-specific RNAs and conserved cellular proteins affect transcription, degrade mRNA transcripts or inhibit translation. Interestingly, S. cerevisiae and its closest relatives lack the genes required for RNAi, yet maintain tight control over Ty1 retrotransposition. We discovered a copy number control (CNC) system based on Ty1 antisense (AS) RNAs that inhibits retrotransposition posttranslationally. The noncoding transcripts are packaged into virus- like particles and block reverse transcription by greatly decreasing the level of the conserved Ty1 protein integrase, which is required for Ty1 as well as HIV-1 reverse transcription and integration. Our goal is to understand the mechanism of this novel form of RNAi by 1) determining how Ty1AS transcripts inhibit retrotransposition, 2) identifying sequence domains that mediate Ty1AS RNA synthesis, packaging and activity within virus-like particles, and 3) characterizing cellular genes that influence AS transcript synthesis and function. We expect this study to impact two areas of biomedical research. First, understanding Ty1 CNC can be used to help elucidate the function of HIV noncoding RNAs, improve RNA-based therapies, and identify new targets for drug development. Second, our work will inform research addressing how noncoding RNAs impact human gene expression on a broader scale, especially when one considers that over 90% of the genome is transcribed, yet the function of many of these transcripts is poorly understood. PUBLIC HEALTH RELEVANCE: Understanding the mechanism of a novel RNA-based defense against the retroviral-like transposable element Ty1 will inform areas of biomedical research concerned with elucidating the function of HIV noncoding RNAs, improving RNA-therapies to combat HIV/AIDS and identifying new targets for drug development. In addition, our work will yield insight into how noncoding RNAs impact human gene expression on a broader scale, especially when one considers that most of the genome is transcribed, yet the role many of these transcripts play in health and disease is poorly understood.

描述（由申请人提供）：反义RNA控制逆转录室拷贝编号酿酒酵母的类似逆转录病毒的转座子TY1是理解HIV的复制的有效模型，艾滋病的复制，艾滋病的病原体，可以说是最良好的理解的转子培训。逆转录座子通过RNA中间体复制，并在大多数真核基因组定殖。研究TY1还提供了有关人类基因组动力学和进化以及转座子诱导的突变的行为的宝贵信息。 RNA干扰（RNAi）是一种广泛的机制，用于沉默传染病和可转座元件，并改变基因表达，其中小序列特异性RNA和保守的细胞蛋白会影响转录，降解mRNA转录本或抑制翻译。有趣的是，酿酒酵母及其最接近的亲戚缺乏RNAi所需的基因，但仍保持对TY1逆转录的严格控制。我们基于TY1反义（AS）RNA发现了一个拷贝数控制（CNC）系统，该系统会在翻译后抑制逆转录位置。通过大大降低保守的TY1蛋白积分酶的水平，将非编码转录物包装到病毒中，并阻止逆转录，这是TY1以及HIV-1逆转录和整合所必需的。我们的目标是通过1）确定TY1AS转录本如何抑制逆转录置位的方法来了解这种新型RNAi的机制，2）识别介导Ty1AS RNA合成，病毒样颗粒中的TY1AS RNA合成，包装和活性的序列域，以及3）表征影响转录物综合和功能的细胞基因。 我们希望这项研究会影响生物医学研究的两个领域。首先，了解TY1 CNC可用于帮助阐明HIV非编码RNA的功能，改善基于RNA的疗法并确定药物开发的新目标。其次，我们的工作将为无编码的RNA在更广泛的范围内影响人类基因表达的研究提供信息，尤其是当人们认为超过90％的基因组被转录时，但许多这些转录本的功能却鲜为人知。 公共卫生相关性：了解针对逆转录病毒的可转移元件Ty1的新型防御机制将为您提供有关阐明HIV非编码RNA功能的生物医学研究领域的信息，从而改善了RNA触及型RNA治疗，从而对抗HIV/AIDS进行对抗HIV/AIDS，并确定药物开发的新目标。此外，我们的工作将深入了解不编码的RNA如何在更广泛的范围内影响人类基因的表达，尤其是当人们认为大多数基因组都被转录时，但这些转录中许多转录物在健康和疾病中的作用却很少。