A Novel RNA Polyphosphatase PIR-1 Plays Important Roles in Silencing Viruses and Regulating Genes in C. elegans

一种新型 RNA 多磷酸酶 PIR-1 在沉默病毒和调节秀丽隐杆线虫基因中发挥重要作用

• 批准号: 10361459
• 负责人: Weifeng Gu
• 金额: $ 32.66万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361459
• 关键词: Affect; Animals; Antiviral Response; Antiviral Therapy; Binding; Biogenesis; Biological; Biological Process; Biology; Biomedical Research; Caenorhabditis elegans; Cancer Biology; Cell Culture Techniques; Cell Proliferation; Cells; Communicable Diseases; Complex; Data; Defect; Development; Developmental Biology; Disease; Double-Stranded RNA; Enzymes; Fertility; Gene Expression; Gene Expression Regulation; Genes; Genetic; Human; In Vitro; Lead; Life; Malignant Neoplasms; Mediating; Messenger RNA; MicroRNAs; Modeling; Molecular; Nobel Prize; Normal Cell; Nucleotides; Pathologic; Pathway interactions; Phenotype; Physiological; Play; Process; Proteins; Public Health; RNA; RNA Binding; RNA Caps; RNA Interference; RNA Interference Pathway; RNA Precursors; RNA Splicing; RNA Stability; RNA-Directed RNA Polymerase; Regulation; Research; Role; Scientific Inquiry; Small Interfering RNA; Specific qualifier value; System; Textbooks; Therapeutic; Viral; Virus; Virus Diseases; Work; anti-cancer; base; field study; human disease; in vivo; innovation; inorganic phosphate; insight; mRNA capping; mRNA guanylyltransferase; mutant; neoplastic cell; novel; protein expression; recruit; tool; transcription factor; viral RNA; virology

Project Summary/Abstract Dysregulation in gene expression is involved in most human diseases including cancers, developmental defects, and viral infections. Many therapeutic tools target defects in gene expression. The PI will study a key step in gene regulation – 'how nascent triphosphorylated RNAs (ppp-RNAs) are regulated?'. RNA interference (RNAi) plays important roles in regulating genes, including viral and 'harmful' genes in all domains of life. This project will systematically examine how a highly conserved RNA polyphosphatase, C. elegans PIR-1, regulates gene expression by modifying ppp-RNAs in RNAi and non-RNAi-mediated pathways. In textbooks, nascent RNAs are synthesized as ppp-RNAs, which, as gene expression intermediates, are immediately converted to capped RNAs for protein expression or other functions. This study will demonstrate how an RNA polyphosphatase can change the fate of nascent ppp-RNAs, thereby modifying the textbook model. This gene regulation mechanism will serve as a new tool to regulate genes in research and therapeutics. Previous studies have indicated that human PIR-1 (hPIR1), a novel RNA polyphosphatase, removes two phosphates from ppp-RNAs to generate p-RNAs in vitro, which are then degraded. This activity renders PIR-1 a candidate for regulating ppp-RNAs including nascent mRNAs and viral RNAs. Consistently, previous studies suggested hPIR1 may be involved in regulating specific genes critical for cell proliferation in normal and tumor cells; however, these studies have provided few insights into: 1) what are the targets genes of PIR-1; 2) how PIR-1 works in vivo. The PI has identified two types of RNA substrates for C. elegans PIR-1, viral RNAs and endogenous mRNAs, both of which are silenced by PIR-1-mediated RNAi pathways. These preliminary results are consistent with the previous observation that PIR-1 may interact with Dicer, a central component of RNAi. Although the mechanism of RNAi has been extensively examined, previous studies have provided few insights into whether the ppp group on viral and cellular double-stranded (ds)RNAs plays any role in RNAi. The PI proposes PIR-1 may modify ppp-dsRNAs to generate p-dsRNAs, promoting Dicer to efficiently cut dsRNAs in C. elegans. The PI will systematically dissect the molecular mechanism of how PIR-1 regulates genes in these RNAi processes. The PI will also examine how PIR-1 regulates nascent ppp-RNAs in non-RNAi processes which play essential roles in the fertility of C. elegans. C. elegans is a robust genetic system especially in RNAi fields, and studies using C. elegans have led to several breakthroughs including three Nobel prizes in the last decade. The proposed research is original and innovative since it examines an underexplored yet key gene regulation mechanism. This research may lead to the development of more effective tools for regulating viral and endogenous genes. This project will have a significant and long-lasting impact on broad scientific inquiries including RNA biology, virology, developmental biology, and cancer biology. This study will also provide a new gene regulation tool for anti-cancer and anti-viral therapies, thus exerting a significant impact on public health.

项目总结/摘要 基因表达的失调涉及大多数人类疾病，包括癌症、发育性疾病、遗传性疾病、免疫缺陷综合征和免疫缺陷综合征。 缺陷和病毒感染。许多治疗工具靶向基因表达缺陷。私家侦探会研究一把钥匙 基因调控的步骤-新生的三磷酸化RNA（ppp-RNA）是如何被调控的？'. RNA干扰 RNA干扰（RNAi）在调节基因中起着重要作用，包括生命所有领域中的病毒和“有害”基因。这 该项目将系统地研究高度保守的RNA多磷酸酶C.线虫PIR-1，调节 在RNAi和非RNAi介导的途径中通过修饰ppp-RNA进行基因表达。在教科书中，新生的 RNA被合成为ppp-RNA，作为基因表达的中间产物，ppp-RNA被立即转化为 用于蛋白质表达或其他功能的加帽RNA。这项研究将证明RNA如何 多聚磷酸酶可以改变新生ppp-RNA的命运，从而修改教科书模型。该基因 调控机制将成为研究和治疗基因调控的新工具。先前 研究表明，人PIR-1（hPIR 1）是一种新的RNA多磷酸酶， 在体外从ppp-RNA产生p-RNA，然后降解。这项活动使PIR-1成为候选者 用于调节ppp-RNA，包括新生mRNA和病毒RNA。一致的是，先前的研究表明， hPIR 1可能参与调节正常和肿瘤细胞中对细胞增殖至关重要的特定基因; 然而，这些研究提供了很少的见解：1）PIR-1的靶基因是什么; 2）PIR-1如何在细胞内表达， 在活体内有效PI已经鉴定了C的两种类型的RNA底物。线虫PIR-1、病毒RNA和 内源性mRNA，这两者都是通过PIR-1介导的RNAi途径沉默的。这些初步结果 与先前的观察结果一致，即PIR-1可能与RNAi的核心组分Dicer相互作用。 虽然RNAi的机制已被广泛研究，但以前的研究提供的见解很少 病毒和细胞双链（ds）RNA上的ppp基团是否在RNAi中发挥作用。的PI 提出PIR-1可以修饰ppp-dsRNA以产生p-dsRNA，促进Dicer有效地切割dsRNA， C.优雅的PI将系统地剖析PIR-1如何调节这些基因的分子机制， RNAi过程。PI还将研究PIR-1如何在非RNAi过程中调节新生ppp-RNA 在C.优雅的C.线虫是一个强大的遗传系统，特别是在RNAi中， 领域，并使用C.在过去的几年里，包括三次诺贝尔奖在内的几项突破， 十年这项研究是原创性和创新性的，因为它研究了一个尚未探索的关键基因。 调节机制。这项研究可能会导致开发更有效的工具来调节病毒 和内源基因。该项目将对广泛的科学调查产生重大而持久的影响 包括RNA生物学、病毒学、发育生物学和癌症生物学。这项研究还将提供一个新的 基因调控工具，用于抗癌和抗病毒治疗，从而对公众健康产生重大影响。