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和疾病的发展无关。腺病毒于1953年首次从扁桃体中鉴定出来。1968年，美国武装部队的急性呼吸道疾病委员会发现腺病毒感染是新兵急性急性呼吸道疾病的主要原因之一。在过去的五十年中，腺病毒研究对病毒性肺炎，疫苗接种以及分子病毒学和细胞生物学的基本方面的发病机理产生了令人印象深刻的知识。但是，腺病毒仍然是一个重大的平民和军事威胁。发现腺病毒受体（Coxsackievivirus和腺病毒受体； CAR），最近发现汽车在气道上皮细胞粘附中起着核心作用，从而产生了从上皮表面的病毒逃逸的新机制。仍然存在的一个主要问题是，当受体在基底外侧膜上安全分离时，这些致病性病毒如何引发感染。已经描述了几种蛋白质同工型，包括两种跨膜形式（Carex7和Carex8）。这两种同工型仅在其C末端有所不同，这表明它们的某些相互作用及其定位和调节可能有所不同。在极化上皮细胞中，CAREX7位于基底外侧表面，因此隔离了顶部的潜在病毒相互作用。相反，尽管Carex8是一种不太丰富的同工型，但我们最近的工作表明，Carex8是顶端局部的，它可以从根尖表面介导腺病毒感染的开始。我们假设受体丰度和顶端定位受到基于PDZ的相互作用与膜相关的鸟苷酸激酶的相互作用倒入1，同工型B（MAGI-1B）。我们旨在通过PDZ结构域的分离，突变，结合和竞争来理解这种相互作用的分子基础，最终在极化气道细胞中。此外，我们假设与MAGI-1B的相互作用与根尖贩运途径中的含PDZ域蛋白质竞争。我们将采用一种候选蛋白质方法，并通过免疫细胞化学和共免疫沉淀 - 西方印迹分析来研究新的相互作用。由于几个原因，了解顶部定位背后的分子机制在临床上具有重要意义。目前尚无针对Coxsackievievivivirus或腺病毒感染的特定治疗方法。因此，面对病毒爆发时病毒的顶端结合的能力将是一个重大的治疗进展。另一方面，增强受体的顶端表达的能力对于有效的腺病毒介导的基因治疗具有很高的相关性。此外，这项工作将使一支毕业生和本科生团队在病毒和医学的界面上进行重要研究。公共卫生相关性：病毒引起的急性呼吸道疾病每年引起大量的人类疾病和死亡。最近，我们发现了一种结合Coxsackievievirus和腺病毒的受体形式，存在于气道上皮的空气表面上。了解原因是什么原因导致该表面位置以及如何调节这将提供对病毒感染易感性的洞察力，并导致预防病毒感染和促进腺病毒介导的基因治疗的策略，以治疗遗传和获得的呼吸道疾病。