Peptidyl-prolyl isomerase Pin1 as a multifunctional regulator of cytomegalovirus replication

肽基脯氨酰异构酶 Pin1 作为巨细胞病毒复制的多功能调节剂

• 批准号: 341613338
• 负责人: Professor Dr. Manfred Marschall, since 9/2018
• 金额: --
• 依托单位: Virologisches Institut - Klinische und Molekulare Virologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/341613338?language=en
• 关键词: Peptidyl; prolyl; isomerase; Pin1; multifunctional

Virus replication is a complex process in general that includes multiple steps of intracellular transport and virion morphogenesis as determined by intense virus-host interaction. In this aspect, recent reports suggested that cellular peptidyl-prolyl isomerases (PPIases) represent a novel group of enzymes that are crucial for efficient replication of various viruses and might be more important than previously expected. In case of herpesviruses, our group has recently provided the first evidence that the cellular isomerase Pin1 facilitates nuclear egress of cytomegaloviral particles. This consequently opens up the intriguing possibility that PPIases might be determinants of virus-host interaction and might even be utilized as novel targets for antiherpesviral strategies. In this project, we will extend our understanding of the role of protein prolyl-isomerization in the regulation of herpesvirus infection. In particular, we intend to characterize important regulatory functions of the PPIase Pin1 during replication of the human and mouse cytomegalovirus. We will address the following questions: (i) is Pin1 activity required for the regulation of distinct steps of the viral replication cycle besides nuclear egress, (ii) which regulatory proteins are subject to Pin1-mediated isomerization in virus-infected cells, and (iii) what are the molecular mechanisms controlling Pin1 activity by the virus? We will additionally develop a microscopy-based assay enabling the analysis of prolyl-isomerization for the first time in living cells. To address these issues, we will use various state-of-the-art techniques including super-resolution STED/RESOLFT microscopy, nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. The presentedproject will be supported by already fruitful ongoing collaborations and will particularly benefit from the recruitment of further designated experts in their fields. The work programme will specifically help to validate PPIases as novel targets for treatment of herpesviral infections and to develop strategies for future therapeutic interventions.

病毒复制通常是一个复杂的过程，包括细胞内转运和病毒体形态发生的多个步骤，这是由强烈的病毒-宿主相互作用决定的。在这方面，最近的报告表明，细胞肽基脯氨酰异构酶（PPIases）代表了一组新的酶，是各种病毒的有效复制的关键，可能比以前预期的更重要。在疱疹病毒的情况下，我们的小组最近提供了第一个证据，细胞异构酶Pin 1促进巨细胞病毒颗粒的核出口。因此，这开辟了有趣的可能性，PPIases可能是病毒-宿主相互作用的决定因素，甚至可能被用作抗疱疹病毒策略的新靶点。在这个项目中，我们将扩展我们对蛋白质脯氨酰异构化在疱疹病毒感染调节中的作用的理解。特别是，我们打算在人类和小鼠的巨细胞病毒复制过程中的PPIase Pin 1的重要调控功能的特点。我们将解决以下问题：（一）是Pin 1活动所需的调节不同的步骤，病毒复制周期除了核出口，（二）哪些调节蛋白受到Pin 1介导的异构化病毒感染的细胞，和（iii）是什么样的分子机制控制Pin 1活性的病毒？我们还将开发一种基于显微镜的测定方法，首次在活细胞中分析脯氨酰异构化。为了解决这些问题，我们将使用各种最先进的技术，包括超分辨率STED/RESOLFT显微镜，核磁共振（NMR）光谱和质谱。本项目将得到已经富有成效的持续合作的支持，并将特别受益于在其领域进一步招聘指定专家。该工作计划将具体帮助验证PPIases作为治疗疱疹病毒感染的新靶点，并制定未来治疗干预措施的策略。