Initiation and regulation of RSV mRNA transcription and genome replication

RSV mRNA 转录和基因组复制的启动和调节

• 批准号: 8767626
• 负责人: Rachel Fearns
• 金额: $ 41.5万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-07 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8767626
• 关键词: Acute; Address; Antiviral Agents; Area; Asthma; Biochemical Genetics; Biological Assay; Caring; Cessation of life; Characteristics; Child; Chronic Obstructive Airway Disease; Clinical; Complement; Complex; Cost of Illness; Cryoelectron Microscopy; Data; Development; Dinucleoside Phosphates; Disease; Drug Targeting; Elderly; Elements; Future; Gene Expression; Genetic Transcription; Genome; Goals; Hospitalization; Immunity; Immunocompromised Host; In Vitro; Individual; Infant; Intensive Care; Ligation; Lung diseases; Medical; Messenger RNA; Methodology; Molecular; Molecular Biology; Multienzyme Complexes; Negative Staining; Nucleocapsid; Nucleotides; Patients; Pharmaceutical Preparations; Phosphoproteins; Polymerase; Process; Production; Promoter Regions; Property; Proteins; Public Health; RNA; RNA chemical synthesis; RNA replication; RNA-Directed RNA Polymerase; RNA-Protein Interaction; Recombinants; Regulation; Replication Initiation; Research; Resolution; Respiration Disorders; Respiratory Syncytial Virus Infections; Respiratory Tract Diseases; Respiratory Tract Infections; Respiratory syncytial virus; Respiratory syncytial virus RSV proteins; Severities; Site; Small RNA; Source; Structural Biologist; Structure; Supportive care; Time; Transcriptase; Transcription Initiation; United States; Vaccines; Viral; Viral Genes; Viral Load result; Virus; Virus Diseases; Work; analog; design; genetic regulatory protein; improved; innovation; insight; mortality; positional cloning; promoter; public health relevance; recombinant RNA; replicase; research study; small molecule; structural biology; viral RNA

DESCRIPTION (provided by applicant): Respiratory syncytial virus (RSV) is a significant public health concern. It is the major cause of respiratory disease in infants. It also causes severe disease in individuals who are immunocompromised, elderly or have underlying lung disorders, such as asthma or chronic obstructive pulmonary disease. Currently there is no vaccine or effective antiviral drug to treat patients infected with RSV. Treatment is limited to supportive care, which in severe cases requires hospitalization and intensive care. Thus, there is a clear need for anti-RSV drugs that limit the severity of RSV disease to reduce the mortality associated with RSV, as well as the need for intensive treatment. An ideal target for antiviral drugs is the RSV RNA dependent RNA polymerase (RdRp), as it has unique and essential enzymatic activities. The RdRp transcribes and replicates the non-segmented negative sense RNA genome and is capable of initiating both activities from the same viral promoter. The goal of this project is to generate detailed molecular information about key functional and structural properties of the RSV RdRp. The work is divided into three major aims. The experiments in Aim 1 will elucidate the mechanisms by which the RdRp initiates transcription and RNA replication from two sites within the same promoter element, and determine the likelihood of using small molecules to successfully inhibit these steps. Aim 2 will determine how the RdRp is assembled into fully-competent transcriptase and replicase complexes, and will investigate how these complexes are regulated. The research in Aim 3 will utilize negative stain and cryo-electron microscopy to gain insight into the structural characteristics of the RdRp alone and in association with promoter RNA and the nucleocapsid. This research will significantly improve our understanding of the functional and structural properties of the RdRp and provide insights that could be exploited for rational development of antiviral drugs.

描述（由申请人提供）：呼吸综合病毒（RSV）是一个重大的公共卫生问题。它是婴儿呼吸道疾病的主要原因。这也会引起免疫功能低下，老年人或潜在肺部疾病（例如哮喘或慢性阻塞性肺疾病）的患者的严重疾病。目前没有疫苗或有效的抗病毒药来治疗感染RSV的患者。治疗仅限于支持性护理，在严重的情况下需要住院和重症监护。因此，显然需要抗RSV药物，这些抗RSV药物限制了RSV疾病的严重程度降低与RSV相关的死亡率以及对强化治疗的需求。 RSV RNA依赖性RNA聚合酶（RDRP）的理想靶标是它具有独特且必不可少的酶促活性，因此RSV RNA依赖性RNA聚合酶（RDRP）。 RDRP转录并复制非细分的负性RNA基因组，并能够启动来自同一病毒启动子的两种活性。该项目的目的是生成有关RSV RDRP的关键功能和结构特性的详细分子信息。这项工作分为三个主要目标。 AIM 1中的实验将阐明RDRP从同一启动子元素内的两个位点启动转录和RNA复制的机制，并确定使用小分子成功抑制这些步骤的可能性。 AIM 2将确定如何将RDRP组装到完全竞争的转录酶和复制酶配合物中，并将研究这些复合物的调节方式。 AIM 3中的研究将利用阴性染色和冷冻电子显微镜，以深入了解单独的RDRP的结构特征，以及与启动子RNA和核壳的结合特征。这项研究将显着提高我们对RDRP功能和结构特性的理解，并提供可以利用抗病毒药药理性发展的见解。