A systems approach to elucidation of protein lipidation during virus infection

阐明病毒感染过程中蛋白质脂化的系统方法

• 批准号: MR/L000148/1
• 负责人: Peter O'Hare
• 金额: $ 50.1万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FL000148%2F1
• 关键词: systems; approach; elucidation; protein; lipidation

Viruses are small infectious agents and are obligate parasites in that they replicate only inside host cells. To cause disease, they must enter specific target cells, overcome barriers to infection and engage with the appropriate cellular machinery which they redirect to make more copies of themselves. This application concerns one of a major group of human viruses, the herpesviruses. These viruses cause diseases ranging from moderate but with high frequency in the population, such as cold sores, chicken pox or glandular fever, through to diseases that while less frequent, can be extremely severe and life threatening, including eye infections, diseases in newborn infants, as well as peripheral and central nervous system diseases. They are particularly dangerous in immunosuppressed patients such as transplant recipients and in AIDS patients. While we have some effective drugs, there are significant limitations with their use and they are effective only against a subset of the herpesviruses. To identify new therapeutic opportunities we must have a thorough understanding of the mechanisms of replication. Such understanding may lead to a rationale way to design novel antiviral agents which may be more broadly effective. Host cell proteins and viral proteins are modified after (or during) their synthesis with a series of different types of molecules. This process is generally termed post-translational modification and it plays a fundamental role in changing the structure, localisation, stability and function of the potential target proteins. Therefore in terms of virus infection, protein modification can be on host cell proteins and affect their ability to support virus growth or to respond to infection and mount an antiviral attack. Host protein modification can in principle remain unaffected, or specific species can be induced by the host, or by the virus, while other species may be down-regulated. Likewise modification of virus proteins can affect their function, or recruitment into assembly particles, their transport within and out of a cell or their ability to overcome host responses. One general type of such modification is called lipidation, wherein proteins are covalently coupled to any of a series of lipids (fat like molecules). This represents a key regulatory mechanism in altering proteins and thus regulating processes including membrane targeting, intracellular protein and organelle transport, signalling networks relaying communication from outside to inside the cell, cell structure, metabolic pathways, neurotransmission and immune responses. As such protein lipidation plays a profound role in normal biological functioning and disease progression. There are four broad types of lipid modification and they can take place on proteins in different combinations, altering the location and function of proteins. However lipidation is broadly less understood than other sorts of protein modification and our knowledge of the range of host and viral proteins which are modified is incomplete. This is because the most frequently utilised approaches lack sensitivity, can be slow to yield results and are not readily amenable to high-throughput analysis exploiting advances in genetic and protein bioinformatics. We aim to develop novel chemical technologies combined with high-throughput approaches on the role of lipidation of virus and host proteins in herpesvirus infection. Not only will this work yield novel insight into the identity and role of lipidation in this context, it will produce new probes and methodologies available for other systems. It will hugely accelerate our understanding of infection and our ability to perform similar studies in other disease systems not only in infectious diseases. It will enable high-throughput screening of potential new classes of therapeutics and it will provide a roadmap for multidisciplinary working bringing together chemistry and biology to yield novel insight into disease processes.

病毒是小型传染性病原体，并且是专性寄生虫，因为它们仅在宿主细胞内复制。为了引起疾病，它们必须进入特定的靶细胞，克服感染障碍，并与适当的细胞机制接触，它们重新定向以制造更多的自身拷贝。本申请涉及人类病毒的主要组之一，疱疹病毒。这些病毒引起的疾病范围从中度但在人群中频率很高的疾病，如唇疱疹、水痘或腺热，到频率较低但可能极其严重和危及生命的疾病，包括眼部感染、新生儿疾病以及外周和中枢神经系统疾病。它们在免疫抑制患者如移植受体和艾滋病患者中特别危险。虽然我们有一些有效的药物，但它们的使用有很大的局限性，它们只对疱疹病毒的一个子集有效。为了确定新的治疗机会，我们必须彻底了解复制的机制。这样的理解可能会导致一个合理的方式来设计新的抗病毒药，可能是更广泛的有效。宿主细胞蛋白质和病毒蛋白质在其合成之后（或期间）被一系列不同类型的分子修饰。这个过程通常被称为翻译后修饰，它在改变潜在靶蛋白的结构、定位、稳定性和功能方面起着重要作用。因此，就病毒感染而言，蛋白质修饰可能发生在宿主细胞蛋白上，并影响其支持病毒生长或对感染做出反应并发起抗病毒攻击的能力。宿主蛋白质修饰原则上可以保持不受影响，或者特定物种可以由宿主或病毒诱导，而其他物种可以下调。同样，病毒蛋白的修饰可以影响它们的功能，或募集到组装颗粒中，它们在细胞内和细胞外的运输或它们克服宿主反应的能力。这种修饰的一种一般类型称为脂化，其中蛋白质共价偶联到一系列脂质（脂肪样分子）中的任一种。这代表了改变蛋白质的关键调节机制，从而调节包括膜靶向、细胞内蛋白质和细胞器运输、从细胞外到细胞内传递通信的信号网络、细胞结构、代谢途径、神经传递和免疫应答的过程。因此，蛋白质脂化在正常生物功能和疾病进展中起着重要作用。有四种广泛类型的脂质修饰，它们可以以不同的组合发生在蛋白质上，改变蛋白质的位置和功能。然而，脂化与其他种类的蛋白质修饰相比被广泛地理解得较少，并且我们对被修饰的宿主和病毒蛋白质的范围的知识是不完整的。这是因为最常用的方法缺乏灵敏度，产生结果的速度可能很慢，并且不容易适用于利用遗传和蛋白质生物信息学进展的高通量分析。我们的目标是开发新的化学技术结合高通量的方法在疱疹病毒感染的病毒和宿主蛋白质的脂质化的作用。这项工作不仅将产生新的洞察力的身份和作用的脂质化在这种情况下，它将产生新的探针和方法可用于其他系统。它将极大地加速我们对感染的理解，以及我们在其他疾病系统中进行类似研究的能力，而不仅仅是在传染病中。它将能够高通量筛选潜在的新类别的治疗方法，并将为多学科工作提供路线图，将化学和生物学结合在一起，以产生对疾病过程的新见解。

项目成果

Herpes Simplex Virus 1 (HSV-1) ICP22 protein directly interacts with cyclin-dependent kinase (CDK)9 to inhibit RNA polymerase II transcription elongation.

• DOI: 10.1371/journal.pone.0107654
• 发表时间: 2014
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Zaborowska J;Baumli S;Laitem C;O'Reilly D;Thomas PH;O'Hare P;Murphy S
• 通讯作者: Murphy S

Global analysis of protein N-myristoylation and exploration of N-myristoyltransferase as a drug target in the neglected human pathogen Leishmania donovani.

• DOI: 10.1016/j.chembiol.2015.01.003
• 发表时间: 2015-03-19
• 期刊: Chemistry & biology
• 作者: Wright MH;Paape D;Storck EM;Serwa RA;Smith DF;Tate EW
• 通讯作者: Tate EW

Remote Activation of Host Cell DNA Synthesis in Uninfected Cells Signaled by Infected Cells in Advance of Virus Transmission.

• DOI: 10.1128/jvi.01950-15
• 发表时间: 2015-11
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Schmidt N;Hennig T;Serwa RA;Marchetti M;O'Hare P
• 通讯作者: O'Hare P

Systems Analysis of Protein Fatty Acylation in Herpes Simplex Virus-Infected Cells Using Chemical Proteomics.

• DOI: 10.1016/j.chembiol.2015.06.024
• 发表时间: 2015-08-20
• 期刊: Chemistry & biology
• 作者: Serwa RA;Abaitua F;Krause E;Tate EW;O'Hare P
• 通讯作者: O'Hare P

Multifunctional reagents for quantitative proteome-wide analysis of protein modification in human cells and dynamic profiling of protein lipidation during vertebrate development.

• DOI: 10.1002/anie.201500342
• 发表时间: 2015-05-11
• 期刊: ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
• 影响因子: 16.6
• 作者: Broncel, Malgorzata;Serwa, Remigiusz A.;Ciepla, Paulina;Krause, Eberhard;Dallman, Margaret J.;Magee, Anthony I.;Tate, Edward W.
• 通讯作者: Tate, Edward W.