Biology of RNA G-quadruplexes

RNA G 四链体的生物学

基本信息

批准号：
10093075
负责人：
Pavel Ivanov
金额：
$ 37.59万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10093075
关键词：
3-Dimensional 7-deazaguanine Affect Base Pairing Binding Binding Proteins Biochemical Biological Biological Assay Biological Process Biological Testing Biology Biophysics Cancer Biology Cell physiology Cells Cellular Stress Collaborations DNA Data Development Disease Elements Etiology Eukaryotic Cell Event Functional disorder Gene Expression Genes Genetic Transcription Genetic Translation Goals Growth Guanine Guanosine Homeostasis Human In Vitro Intervention Investigation Ions Laboratories Lead Malignant Neoplasms Mediating Messenger RNA Metabolism Methods Molecular Molecular Target Mutation Nerve Degeneration Neurodegenerative Disorders Nucleic Acid Regulatory Sequences Nucleic Acids Pathologic Pathology Pathway interactions Pharmacology Physiological Poly A Process Proteins Proteomics Publishing RNA RNA Binding RNA Sequences RNA metabolism Regulation Regulatory Element Reporting Research Ribonucleic Acid Regulatory Sequences Role Stress Structure System Techniques Testing Therapeutic Transcript Translations Work base biological adaptation to stress biophysical techniques genetic manipulation genome-wide analysis human disease in vivo interest mRNA Stability novel novel strategies novel therapeutics physiologic stressor prevent telomere three dimensional structure transcriptome tumorigenic

项目摘要

Summary G-quadruplexes (G4s) are non-canonical secondary structures in nucleic acids that are formed by guanine-rich sequences. G4 structural and functional studies have largely focused on DNA G4s, and the number of biological functions assigned to these motifs has grown rapidly since the discovery of their involvement in telomere biology. RNA G4s (RG4s) are less studied, but interest is increasing due to their association with multiple processes. A comprehensive understanding of how RNA G4s contribute to cell physiology and pathophysiology is the long-term research goal of the applicant. Multiple reports clearly demonstrate that G4s are enriched in mRNA 5’- and 3’-untranslated regulatory regions. There is an increasing evidence that RG4s control gene expression at transcriptional and post- transcriptional levels, although such data is largely based on in vitro studies. Testing the biological significance of RG4s requires proving that RG4s exist in vivo. Intriguingly, RG4s appear predominantly unfolded in eukaryotic cells, whereas they are readily folded in vitro, suggesting that in cells RG4s are constitutively recognized and actively unfolded. We hypothesize that RG4 folding-unfolding regulates mRNA homeostasis. This hypothesis is based on our analysis of human transcriptome that identifies RG4s as stress-responsive RNA elements. We will test this hypothesis with three specific aims. In AIM1, we will determine and characterize the fraction of the human transcriptome that contains putative stress-responsive RG4s in living cells. In AIM2, we will identify bona fide RG4-binding proteins using a novel approach based on proteomic/biochemical analysis of interactions between the 7-deazaguanine RNA derivatives and proposed binding factors. In AIM 3, we will use functional assays to determine the biological significance of RG4s in mRNA stability, localization and translation. We will use biophysical and biochemical methods to validate selected RG4 candidates. This work will elucidate how RG4-mediated functions contribute to cellular mRNA homeostasis, and will identify physiologically significant RG4-binding partners, which in turn may reveal molecular targets and pathways with therapeutic potential. The understanding of cellular functions of RG4 motifs is particularly relevant to the biology of cancer and neurodegeneration. RNA regions containing RG4s significantly overlap with regions containing disease- associated mutations. The proposed studies may elucidate molecular events underlying normal and pathological aspects of cell physiology, and identify events contributing to the tumorigenic or neurodegenerative changes.

概括 g四链体（G4S）是核酸中非经典的二级结构，由核酸形成。鸟嘌呤富含序列。 G4结构和功能研究主要集中在DNA G4上，而自从发现它们以来，分配给这些主题的生物学功能的数量已迅速增长 RNA G4S（RG4S）的研究较少，但由于它们的兴趣正在增加与多个过程相关联。对RNA G4如何贡献细胞的全面理解生理和病理生理学是申请人的长期研究目标。多个报告清楚地表明，G4富集在mRNA 5'-和3'-非翻译调节中地区。越来越多的证据表明，RG4S控制转录和后转录的基因表达转录水平，尽管这些数据主要基于体外研究。测试生物学 RG4需要提供体内RG4。有趣的是，RG4似乎主要展开真核细胞，而它们很容易在体外折叠，表明在细胞中，RG4始终如一公认并积极展开。我们假设RG4折叠无折叠可以调节mRNA稳态。该假设基于我们对人类转录组的分析，该分析将RG4识别为应力响应性 RNA元素。我们将以三个特定的目标检验这一假设。在AIM1中，我们将确定和表征人类转录组的比例，其中包含推定的应力响应RG4 细胞。在AIM2中，我们将使用基于基于新颖的方法来确定真正的RG4结合蛋白蛋白质组学/生化分析7-二氮氨酸RNA衍生物和提议之间的相互作用结合因子。在AIM 3中，我们将使用功能测定来确定RG4在 mRNA稳定性，定位和翻译。我们将使用生物物理和生化方法来验证选定的RG4候选人。这项工作将阐明RG4介导的功能如何促进细胞mRNA 稳态，并将确定身体上重要的RG4结合伙伴，这可能揭示具有治疗潜力的分子靶标和途径。对RG4基序的细胞功能的理解与癌症的生物学特别相关和神经变性。含有RG4的RNA区域与含有疾病的区域显着重叠相关突变。拟议的研究可能阐明正常正常和的分子事件细胞生理学的病理方面，并确定有助于肿瘤性或神经退行性变化。