Understanding the reprogramming of host mRNA translation during calicivirus infection

了解杯状病毒感染期间宿主 mRNA 翻译的重编程

• 批准号: BB/N000943/1
• 负责人: Nicolas Locker
• 金额: $ 44.01万
• 依托单位: University of Surrey
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FN000943%2F1
• 关键词: Understanding; reprogramming; host; mRNA; translation

Our overarching aim is to understand the mechanism by which a group of poorly characterised, yet important viruses, regulate host gene expression to modulate how cells respond to viral infection Cells within the body respond to environmental stimuli and pathogen infection in many ways, the most common of which is via the regulation of cellular gene expression. The expression of the information stored in our genes is tightly controlled at numerous levels, to ensure that the correct protein is produced at the right time and quantity. First, the genetic information encoded in our DNA is converted into messenger RNA (mRNA), via a process called transcription. The mRNA is then "translated" into proteins by structures referred to as ribosomes, assisted by proteins called initiation factors. eIF4E is one of these proteins and its key role is to direct the ribosomes and other proteins to the mRNA. This process is called translation and the synthesised proteins make our cells what they are, defining their properties and functions. Importantly, changes to the proteins in our cells also help to fight against invading pathogens such as viruses. While viruses can infect most organisms and cause severe damage, they consist primarily of RNA or DNA enclosed in a protein coat and lack the factors required for replication and dissemination. They are therefore dependent on the host cell resources to produce viral proteins. Thus, many viruses have developed strategies that regulate the function of the host protein synthesis machinery, often leading to preferentially translation of viral mRNAs. Caliciviruses are a family of small viruses that can cause diseases both in humans and animals. In humans they primarily cause gastroenteritis. While the human norovirus, a calicivirus, does not grow well in cell culture, murine norovirus, a mouse homologue, acts as model with which to study many aspects of calicivirus biology in the laboratory. It represents an excellent model to dissect how viruses affect the translation of host cell proteins. Using this model we have previously made a number of significant advances in the understanding of how the caliciviruses produce viral proteins. We found that a virus-encoded protein, called VPg, is essential for the translation of viral mRNA as it coordinates the recruitment of host proteins to the viral RNA. We also showed that calicivirus infection modulates the composition and activity of the host translation machinery. However, we still know very little about how pathogens in general, and caliciviruses in particular, modulate the translation of host mRNAs during infection. This is important because understanding the modulation of specific host mRNA translation by viruses can reveal how viruses manipulate the organism's response to infection. Our hypothesis is that caliciviruses alter the translation of specific mRNAs in the infected host to regulate antiviral gene expression, and that they do so by modulating the translation factor eIF4E. Therefore, our objectives are to use high-throughput sequencing and biochemical methods to 1- characterize how the viral infection alters the profile of mRNAs that are translated by the infected host and 2- to understand how the activity of eIF4E is regulated by caliciviruses during this process. If we can fully understand how caliciviruses control the activity of translation factors and reprogramme the host protein synthesis, we can identify ways to inhibit virus replication. Therefore, our work will aid in the development of novel antiviral therapies for this important group of viruses, and perhaps other viruses that regulate translation. Understanding the fundamental mechanisms of gene regulation is important not only for virologists but also for broader academic communities. By advancing our basic knowledge of translational control we may understand better several pathologies that are linked to modulation of eIF4E activity, such as cancer and diabetes.

我们的总体目的是了解一组特征性较差但重要的病毒的机制调节宿主基因表达，以调节细胞对体内病毒感染细胞的反应如何对环境刺激和病原体感染的反应在许多方面，其中最常见的是通过调节细胞基因表达的调节。存储在我们基因中的信息的表达在许多水平上受到严格控制，以确保在正确的时间和数量下产生正确的蛋白质。首先，我们DNA中编码的遗传信息通过称为转录的过程将其转换为信使RNA（mRNA）。然后将mRNA“翻译”到蛋白质中，被称为核糖体的结构，并由称为启动因子的蛋白质辅助。 EIF4E是这些蛋白质之一，其关键作用是将核糖体和其他蛋白质引导到mRNA。该过程称为翻译，合成的蛋白质使我们的细胞具有其本质，从而定义其特性和功能。重要的是，我们细胞中蛋白质的变化也有助于抵抗入侵病原体（例如病毒）。虽然病毒会感染大多数生物并造成严重损害，但它们主要由包含在蛋白质外套中的RNA或DNA组成，缺乏复制和传播所需的因素。因此，它们取决于产生病毒蛋白的宿主细胞资源。因此，许多病毒已经开发出调节宿主蛋白质合成机制功能的策略，通常会导致病毒mRNA的优先翻译。彩色病毒是一个小病毒家族，可以引起人类和动物的疾病。在人类中，它们主要引起胃肠炎。虽然人类诺如病毒是一种蜡膜病毒，但在细胞培养中并不能很好地生长，但小鼠同源物鼠莫林病毒是研究实验室中蜡膜病毒生物学许多方面的模型。它代表了一个出色的模型，用于剖析病毒如何影响宿主细胞蛋白的翻译。使用此模型，我们以前在理解蜡膜中如何产生病毒蛋白方面已取得了许多重大进展。我们发现，一种称为VPG的病毒编码的蛋白对于病毒mRNA的翻译至关重要，因为它可以将宿主蛋白募集到病毒RNA中。我们还表明，蜡膜病毒感染调节宿主翻译机制的组成和活性。但是，我们仍然对一般的病原体，尤其是羊轮如何调节感染过程中宿主mRNA的翻译几乎一无所知。这很重要，因为了解病毒对特定宿主mRNA翻译的调节可以揭示病毒如何操纵生物体对感染的反应。我们的假设是，蜡膜病毒改变了受感染宿主中特定mRNA的翻译以调节抗病毒基因表达，并通过调节翻译因子EIF4E来改变。因此，我们的目标是使用高通量测序和生化方法来1-表征病毒感染如何改变受感染宿主翻译的mRNA和2-的mRNA谱，以了解在此过程中eIF4E的活性如何受孔囊病的调节。如果我们能够充分了解蜡囊如何控制翻译因子的活性并重编程宿主蛋白质合成，那么我们可以确定抑制病毒复制的方法。因此，我们的工作将有助于为这一重要病毒以及调节翻译的其他病毒开发新的抗病毒疗法。了解基因调节的基本机制不仅对病毒学家，而且对更广泛的学术社区都很重要。通过促进我们对翻译控制的基本知识，我们可以更好地理解与EIF4E活性调节有关的几种病理，例如癌症和糖尿病。

项目成果

Murine Norovirus Infection Results in Anti-inflammatory Response Downstream of Amino Acid Depletion in Macrophages

• DOI: 10.1128/jvi.01134-21.4
• 发表时间: 2021-10-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Brocard, Michele;Lu, Jia;Locker, Nicolas
• 通讯作者: Locker, Nicolas

Norovirus infection results in eIF2a independent host translation shut-off and remodels the G3BP1 interactome evading stress granule formation

诺如病毒感染导致 eIF2a 独立宿主翻译关闭，并重塑 G3BP1 相互作用组以逃避应激颗粒形成

• DOI: 10.17863/cam.48046
• 发表时间: 2020
• 作者: Brocard M

Murine Norovirus Infection Results in Anti-inflammatory Response Downstream of Amino Acid Depletion in Macrophages.

• DOI: 10.1128/jvi.01134-21
• 发表时间: 2021-09-27
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Brocard M;Lu J;Hall B;Borah K;Moller-Levet C;Georgana I;Sorgeloos F;Beste DJV;Goodfellow IG;Locker N
• 通讯作者: Locker N

Infectious Bronchitis Virus Regulates Cellular Stress Granule Signaling

• DOI: 10.1101/819482
• 发表时间: 2019-10
• 期刊: Viruses
• 作者: M. Brownsword;Nicole Doyle;Michèle Brocard;Nicolas Locker;Helena J. Maier

Norovirus infection results in assembly of virus-specific G3BP1 granules and evasion of eIF2a signaling

诺如病毒感染导致病毒特异性 G3BP1 颗粒组装并逃避 eIF2a 信号传导

• DOI: 10.1101/490318
• 发表时间: 2018
• 作者: Brocard M