Isoform-Specific Regulation and Localization of the Coxsackie and Adenovirus Rece

柯萨奇病毒和腺病毒 Rece 的异构体特异性调控和定位

基本信息

批准号：
7981132
负责人：
Katherine Julie Excoffon
金额：
$ 43.72万
依托单位：
WRIGHT STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-09-22 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7981132
关键词：
Acute Adenovirus Infections Adenoviruses Adhesions Air Apical Arts Belief Binding Biology CAR receptor Cell-Cell Adhesion Cells Cellular biology Cessation of life Chemicals Co-Immunoprecipitations Communication Coxsackie B Viruses Coxsackie Viruses Cystic Fibrosis Transmembrane Conductance Regulator Data Development Disease Disease Outbreaks Endoplasmic Reticulum Environment Epithelial Epithelial Cells Equilibrium Face Family Family member Fever Figs - dietary Golgi Apparatus Grant Hand Human In Vitro Infection Inherited Investigation Knowledge Lead Link Lipids Location Lung Lung diseases Maintenance Mathematics Mediating Medicine Methodology Military Personnel Modeling Molecular Molecular Virology Mutation Pathogenesis Pathway interactions Penetration Play Predisposition Prevention Process Protein Biosynthesis Protein Isoforms Proteins Quality Control Recruitment Activity Regulation Research Role Science Side Site-Directed Mutagenesis Sorting - Cell Movement Structure Students Surface System Therapeutic Tight Junctions Tonsil Training Translating Universities Vaccination Viral Viral Pneumonia Virus Virus Diseases Western Blotting Work adenoviral-mediated adenovirus receptor airway epithelium apical membrane arm base basolateral membrane cell growth regulation clinically significant college gene therapy graduate student immunocytochemistry inhibitor/antagonist insight member membrane-associated guanylate kinase microbial mutant novel polarized cell public health relevance receptor receptor binding receptor expression respiratory sodium-hydrogen exchanger regulatory factor trafficking virology

项目摘要

DESCRIPTION (provided by applicant): Adenovirus and the adenovirus receptor are intricately linked to airway epithelia biology, and to the development of disease. Adenoviruses were first identified from tonsils in 1953. In 1968 the Commission of Acute Respiratory Diseases of the U.S. Armed Forces found that adenovirus infection was one of the major causes of acute febrile respiratory illness in recruits. Over the past 5 decades adenovirus research has yielded impressive knowledge about the pathogenesis of viral pneumonias, vaccination, and basic aspects of molecular virology and cellular biology. However, adenovirus remains a significant civilian and military threat. The discovery of the receptor for adenovirus (Coxsackievirus and adenovirus receptor; CAR) and the recent finding that CAR plays a central role in airway epithelial cell-cell adhesion have yielded novel mechanisms for viral escape from epithelial surfaces. A major unanswered question that remains is how these pathogenic viruses initiate infection when the receptor is safely segregated on the basolateral membrane. Several protein isoforms have been described for CAR, including two transmembrane forms (CAREx7 and CAREx8). These two isoforms differ only in their C-termini, suggesting that some of their interactions, and hence localization and regulation, may differ. In polarized epithelial cells, CAREx7 resides on the basolateral surface and is thus sequestered away from potential viral interactions on the apical surface. In contrast, although CAREx8 is a less abundant isoform, our recent work reveals that CAREx8 is localized apically where it can mediate initiation of adenovirus infection from the apical surface. We hypothesize that receptor abundance and apical localization are regulated by a PDZ-based interaction with membrane-associated guanylate kinase inverted 1, isoform b (MAGI-1b). We aim to understand the molecular basis of this interaction through PDZ domain isolation, mutation, binding and competition, ultimately in polarized airway cells. Moreover, we hypothesize that the interaction with MAGI-1b is in competition with PDZ-domain-containing proteins within the apical trafficking pathway. We will take a candidate protein approach and investigate novel interactions by immunocytochemistry and co-immunoprecipitation-Western blot analysis. Understanding the molecular mechanisms behind apical localization is clinically significant for several reasons. Currently there is no specific treatment for coxsackievirus or adenovirus infection; thus, the ability to block apical binding of the virus in the face of viral outbreaks would be a significant therapeutic advance. On the other hand, the ability to augment apical expression of the receptor would have high relevance for efficient adenoviral-mediated gene therapy. Moreover, this work will expose a team of graduate and undergraduate students to vital research at the interface of virology and medicine. PUBLIC HEALTH RELEVANCE: Viral-induced acute respiratory disease causes a significant amount of human illness and death each year. We have recently discovered one form of the receptor that binds both coxsackievirus and adenovirus is present on the air exposed surface of the airway epithelium. Understanding what causes this surface location and how this is regulated will provide insight into susceptibility to viral infections, and lead to strategies both for prevention of virus infection and facilitation of adenovirus-mediated gene therapy for the treatment of inherited and acquired respiratory diseases.

描述（由申请人提供）：腺病毒和腺病毒受体与气道上皮生物学以及疾病的发展有着复杂的联系。 1953年，腺病毒首次从扁桃体中被发现。1968年，美国武装部队急性呼吸道疾病委员会发现，腺病毒感染是新兵急性发热呼吸道疾病的主要原因之一。过去 5 年来，腺病毒研究在病毒性肺炎的发病机制、疫苗接种以及分子病毒学和细胞生物学的基本方面取得了令人印象深刻的知识。然而，腺病毒仍然是重大的民用和军事威胁。腺病毒受体（柯萨奇病毒和腺病毒受体；CAR）的发现以及最近发现 CAR 在气道上皮细胞粘附中发挥核心作用，为病毒从上皮表面逃逸提供了新的机制。一个尚未解答的主要问题是，当受体安全地隔离在基底外侧膜上时，这些致病病毒如何引发感染。已经描述了 CAR 的几种蛋白质亚型，包括两种跨膜形式（CAREx7 和 CAREx8）。这两种异构体仅在 C 末端不同，表明它们的一些相互作用以及因此的定位和调节可能不同。在极化上皮细胞中，CAREx7 驻留在基底外侧表面，因此与顶端表面上潜在的病毒相互作用隔离。相比之下，虽然 CAREx8 是一种不太丰富的亚型，但我们最近的工作表明 CAREx8 位于顶端，可以介导从顶端表面开始腺病毒感染。我们假设受体丰度和顶端定位是通过基于 PDZ 与膜相关鸟苷酸激酶反向 1，亚型 b (MAGI-1b) 的相互作用来调节的。我们的目标是通过 PDZ 结构域分离、突变、结合和竞争，最终在极化气道细胞中了解这种相互作用的分子基础。此外，我们假设与 MAGI-1b 的相互作用与顶端运输途径中含有 PDZ 结构域的蛋白质竞争。我们将采用候选蛋白质方法，并通过免疫细胞化学和免疫共沉淀-蛋白质印迹分析研究新的相互作用。由于多种原因，了解根尖定位背后的分子机制具有临床意义。目前尚无针对柯萨奇病毒或腺病毒感染的特效治疗方法；因此，在病毒爆发时阻断病毒顶端结合的能力将是一项重大的治疗进展。另一方面，增强受体顶端表达的能力与有效的腺病毒介导的基因治疗具有高度相关性。此外，这项工作将使研究生和本科生团队接触到病毒学和医学交叉领域的重要研究。公共卫生相关性：病毒引起的急性呼吸道疾病每年导致大量人类患病和死亡。我们最近发现一种结合柯萨奇病毒和腺病毒的受体存在于气道上皮暴露于空气的表面上。了解导致该表面位置的原因以及其调节方式将有助于深入了解病毒感染的易感性，并制定预防病毒感染和促进腺病毒介导的基因疗法治疗遗传性和获得性呼吸道疾病的策略。