Structure of origin DNA melting and unwinding complexes of a viral replication protein

病毒复制蛋白的起始 DNA 熔解和解旋复合物的结构

• 批准号: BB/J006920/1
• 负责人: Elena Orlova
• 金额: $ 40.17万
• 依托单位: Birkbeck, University of 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=BB%2FJ006920%2F1
• 关键词: Structure; origin; DNA; melting; unwinding

The papillomaviruses are an important class of disease causing organism in animals and man. Bovine papillomavirus (BPV-1) is the model of the group and the consequences of disease (interference with suckling, milking and breading), has a significant economic impact worldwide. The highly related human viruses (HPV) cause cancer and therefore also have a negative impact on quality of life. The mechanisms of viral replication are consequently of significant interest as the viral replication proteins are important therapeutic targets. The survival and continuity of an organism, be it a virus or man, requires duplication of its genetic material- its DNA- and this is performed by specialized proteins. The first important step in this process is the recognition of specific 'origins of DNA replication' by proteins that separate, or "melt", the two otherwise paired strands of the DNA ('the DNA double helix'). In higher organisms like man, the timing and physical precision of this 'initiation' event is crucial to avoid the catastrophic consequences that may result from any loss of genetic material (for example, cancer). After the initial melting of the DNA at localized site, complete separation of the two helical DNA strands is performed by 'helicase' proteins. DNA strand separation by initiator and helicase proteins is generally poorly understood in simple organisms like viruses as well as higher species like man. However, the process appears to be highly conserved, suggesting that mechanism may be generally similar in all systems. We propose here to explore and characterize these replication processes in BPV-1. The advantage is that all the viral DNA replication activities are performed by a single protein called E1, thus simplifying the study. We will use powerful microscopes to take snapshots of different steps of the process and biochemical techniques that will help to define mechanisms. These studies will therefore contribute significantly to the understanding of the papillomavirus replication protein that is an important therapeutic target. Modeling these replication events in this simplified viral system will also help us understand the process in man. These studies will therefore assist in disease management and improving human and animal health.

牛乳头瘤病毒（BPV-1）是一类重要的动物和人类疾病病原体，是哺乳、挤奶和喂乳过程中的典型疾病，在世界范围内具有重要的经济影响。高度相关的人类病毒（HPV）导致癌症，因此也对生活质量产生负面影响。因此，病毒复制的机制具有重大意义，因为病毒复制蛋白是重要的治疗靶标。一个有机体的生存和延续，无论是病毒还是人，都需要复制其遗传物质--DNA--而这是由专门的蛋白质完成的。这个过程中的第一个重要步骤是识别特定的“DNA复制起点”，通过分离或“融化”DNA的两条配对链（“DNA双螺旋”）的蛋白质。在像人类这样的高等生物中，这个“启动”事件的时间和物理精度对于避免可能因遗传物质的任何损失（例如癌症）而导致的灾难性后果至关重要。在DNA在局部位点的初始解链后，两条螺旋DNA链的完全分离由“解旋酶”蛋白完成。在简单的生物体如病毒以及高等物种如人类中，通过起始子和解旋酶蛋白进行的DNA链分离通常知之甚少。然而，该过程似乎是高度保守的，这表明在所有系统中机制可能大致相似。我们建议在这里探索和表征BPV-1中的这些复制过程。其优点是所有的病毒DNA复制活动都是由一种称为E1的蛋白质完成的，从而简化了研究。我们将使用功能强大的显微镜拍摄过程的不同步骤和生化技术的快照，这将有助于确定机制。因此，这些研究将大大有助于了解乳头瘤病毒复制蛋白，这是一个重要的治疗目标。在这个简化的病毒系统中模拟这些复制事件也将有助于我们理解人类的过程，因此这些研究将有助于疾病管理和改善人类和动物的健康。

项目成果

Structural Study of Heterogeneous Biological Samples by Cryoelectron Microscopy and Image Processing.

通过冷冻显微镜和图像处理对异质生物样品的结构研究。

• DOI: 10.1155/2017/1032432
• 发表时间: 2017
• 期刊: BioMed research international
• 作者: White HE;Ignatiou A;Clare DK;Orlova EV
• 通讯作者: Orlova EV

Structural basis for DNA strand separation by a hexameric replicative helicase.

• DOI: 10.1093/nar/gkv778
• 发表时间: 2015-09-30
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Chaban Y;Stead JA;Ryzhenkova K;Whelan F;Lamber EP;Antson A;Sanders CM;Orlova EV
• 通讯作者: Orlova EV

The ribosome and its role in protein folding: looking through a magnifying glass.

• DOI: 10.1107/s2059798317007446
• 发表时间: 2017-06-01
• 期刊: Acta crystallographica. Section D, Structural biology
• 作者: Javed A;Christodoulou J;Cabrita LD;Orlova EV
• 通讯作者: Orlova EV

DNA induces conformational changes in a recombinant human minichromosome maintenance complex.

• DOI: 10.1074/jbc.m114.622738
• 发表时间: 2015-03-20
• 期刊: The Journal of biological chemistry
• 作者: Hesketh EL;Parker-Manuel RP;Chaban Y;Satti R;Coverley D;Orlova EV;Chong JP
• 通讯作者: Chong JP