Crosstalk between human mRNA nuclear export and polyadenylation machineries

人类 mRNA 核输出和聚腺苷酸化机制之间的串扰

• 批准号: 9195599
• 负责人: NICHOLAS K CONRAD
• 金额: $ 2.94万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9195599
• 关键词: Address; Affect; Antiviral Response; Biochemical; Biogenesis; Biological; Biological Assay; Cell Nucleus; Cell physiology; Cells; Coupled; Coupling; Data; Disease; Down-Regulation; Ensure; Fluorescence; Fluorescence Resonance Energy Transfer; Gene Expression; Genetic; Goals; Human; Image; In Situ Hybridization; In Vitro; Infection; Influenza; Influenza A virus; Length; Link; Malignant Neoplasms; Mammalian Cell; Mediating; Messenger RNA; Microscopy; Molecular; Nuclear; Nuclear Export; Nuclear RNA; Pathogenesis; Pathway interactions; Poly A; Poly(A) Tail; Poly(A)-Binding Proteins; Polyadenylation; Polyadenylation Pathway; Pre-mRNA Polyadenylation Factor; Process; Proteins; Quality Control; RNA Decay; RNA Processing; RNA Stability; Regulation; Resolution; Role; Sum; System; Testing; Tweens; Viral; Virulence Factors; Virus Diseases; Work; base; design; experience; influenza virulence; influenza virus strain; influenzavirus; mRNA Export; mRNA Precursor; messenger ribonucleoprotein; microscopic imaging; mutant; novel; novel therapeutic intervention; prevent; public health relevance; receptor; single molecule

 DESCRIPTION (provided by applicant): Gene expression requires processing of pre-mRNAs prior to the export of mature mRNAs from the nucleus. As a result, cells have evolved mechanisms to ensure that all pre-mRNA processing steps are completed prior to nuclear export. Such mechanisms involve the coupling of mRNA export with upstream steps in mRNA biogenesis, but little is known regarding factors or mechanisms that couple exports with pre-mRNA processing in mammalian cells. We and others have shown that the pathogenic NS1 protein of influenza A virus inhibits mRNA nuclear export, leading to down-regulation of host gene expression and inhibition of host antiviral response. NS1 additionally targets constituents of the polyadenylation machinery. Our preliminary data indicate that NS1 does not affect mRNA export and 3´ processing independently, but instead targets a connection between these steps. Our central goal is to define this novel mechanistic connection between the polyadenylation and mRNA export machineries and uncover how it is targeted by influenza NS1. The application is a multi-PI proposal by Drs. Beatriz Fontoura and Nicholas Conrad, who have extensive expertise in mRNA export and 3´-end formation, respectively. In addition, both PIs have considerable experience with host-viral interactions. In Aim 1, we will determine the role of polyadenylation factors on the regulation of mRNA nuclear export. In Aim 2, we will examine the effects of mRNA export factors on mRNA hyperadenylation and decay. In Aim 3, we will determine the mechanisms of NS1-mediated inhibition of host gene expression at the interface of nuclear export and 3´-end processing. We will use a combination of genetic, biochemical, and cell biological approaches including high-resolution imaging. In sum, we will investigate a novel mechanistic connection between polyadenylation and mRNA nuclear export and demonstrate how it is regulated by the host and by the influenza virus pathogenic factor NS1. Since this work focuses on cellular and viral mechanisms that regulate coupled export and processing and that disregulation of this interface is linked to pathogenesis, these findings may uncover new therapeutic strategies or targets.

 描述（由申请人提供）：基因表达需要在成熟mRNA从细胞核输出之前加工前mRNA。因此，细胞已经进化出机制，以确保在核输出之前完成所有前mRNA加工步骤。这些机制涉及mRNA输出与mRNA生物发生中的上游步骤的偶联，但是关于将输出与哺乳动物细胞中的前mRNA加工偶联的因素或机制知之甚少。我们和其他人已经表明，甲型流感病毒的致病性NS 1蛋白抑制mRNA核输出，导致宿主基因表达下调，抑制宿主抗病毒反应。NS 1还靶向多聚腺苷酸化机制的成分。我们的初步数据表明，NS 1不会独立地影响mRNA输出和3 ′加工，而是靶向这些步骤之间的连接。我们的中心目标是定义多聚腺苷酸化和mRNA输出机制之间的这种新型机制联系，并揭示它如何被流感NS 1靶向。该应用程序是Beatriz Fontoura博士和Nicholas Conrad博士的多PI提案，他们分别在mRNA输出和3 ′端形成方面拥有丰富的专业知识。此外，两名PI均具有相当丰富的宿主-病毒相互作用经验。在目的1中，我们将确定多聚腺苷酸化因子对mRNA核输出调节的作用。在目标2中，我们将研究mRNA输出因子对mRNA高腺苷酸化和衰变的影响。在目标3中，我们将确定NS 1介导的在核输出和3 ′-末端加工界面抑制宿主基因表达的机制。我们将使用遗传，生物化学和细胞生物学方法的组合，包括高分辨率成像。总之，我们将研究多聚腺苷酸化和mRNA核输出之间的一种新的机制联系，并证明它是如何由宿主和流感病毒致病因子NS 1调节的。由于这项工作的重点是细胞和病毒的机制，调节耦合输出和加工，这一接口的失调与发病机制，这些发现可能会发现新的治疗策略或目标。