Functional analysis of the Epstein Barr virus nuclear antigen leader protein (EBNA-LP) in a viral context.

病毒背景下 Epstein Barr 病毒核抗原前导蛋白 (EBNA-LP) 的功能分析。

• 批准号: MR/L008432/1
• 负责人: Robert White
• 金额: $ 57.68万
• 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: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FL008432%2F1
• 关键词: Functional; analysis; Epstein; Barr; virus

Epstein-Barr virus (EBV) infects most of the human population. Infection normally has no symptoms but some people develop infectious mononucleosis (glandular fever) when they become infected. The virus persists life-long in a dormant (latent) state in certain white blood cells. However, occasionally EBV causes cancers of the white blood cells, particularly where individuals are also infected by HIV or Malaria. EBV is also associated with a cancer of the nasopharynx which is unusually common in Southern China. In all, about 1% of all cancers worldwide are thought to involve EBV. The process of establishing EBV latency is governed by some of the viral genes, including the EBV nuclear antigens (EBNAs). The focus of this research grant is one of the EBNAs, EBNA-LP. Previous research has shown that EBNA-LP is important for the ability of EBV to change B cells into a more cancer-like state (transformation): in cell culture EBV can 'transform' B cells to divide indefinitely. EBNA-LP has also been shown to bind to a number of cellular proteins, and to enhance the abilities of another EBNA, EBNA2, to activate certain genes. However, there is no clear understanding of what EBNA-LP does in the context of virus infection. We also don't know whether any of the reported interactions of EBNA-LP with cell proteins are relevant to its functions in EBV infection.To overcome this, I have developed new methods to create EBV strains in which EBNA-LP has been deleted or engineered to contain defined mutations. This was an exceptional technical challenge due to the complex nature of the EBNA-LP gene. Using these viruses, we will be able to assess what roles are played by EBNA-LP in the transformation of B cells, and to identify genes that are controlled by EBNA-LP. I will also clarify the ability of EBNA-LP to interact with cellular proteins, identifying which parts of EBNA-LP bind to these proteins. By correlating the ability of EBNA-LP mutants to interact with a specific host protein and specific defects in transformation, I will identify the biological significance of these interactions and elucidate which ones may provide novel therapeutic targets for EBV. The protein interfaces involved in the interactions and the sites of protein modification may provide targets for chemical compounds that can inhibit those interactions. Revealing precisely what EBNA-LP does for the virus and its mechanism will thus allow future development of assays for novel anti-EBV therapies that may allow elimination of the virus or novel treatments of EBV associated cancers. There are many examples in cancer biology and immune evasion where key cell mechanisms have been discovered by investigation of virus proteins that have evolved to interfere with cell processes. It is therefore hoped that more general insights into disease mechanisms will also come out of the experiments in this research proposal.

eb病毒（EBV）感染了大多数人。感染通常没有症状，但有些人感染后会出现传染性单核细胞增多症（腺热）。这种病毒在某些白细胞中以潜伏状态终生存在。然而，EBV偶尔会引起白细胞癌，特别是当个体也感染了艾滋病毒或疟疾时。EBV还与鼻咽癌有关，这在中国南方非常常见。总的来说，全世界大约1%的癌症被认为与EBV有关。建立EBV潜伏期的过程是由一些病毒基因控制的，包括EBV核抗原（ebna）。这项研究资助的重点是ebna之一，EBNA-LP。先前的研究表明，EBNA-LP对于EBV将B细胞转变为更像癌症的状态（转化）的能力很重要：在细胞培养中，EBV可以“转化”B细胞无限分裂。EBNA- lp也被证明与许多细胞蛋白结合，并增强另一种EBNA EBNA2激活某些基因的能力。然而，对于ena - lp在病毒感染中的作用，目前还没有明确的认识。我们也不知道报道的EBNA-LP与细胞蛋白的相互作用是否与其在EBV感染中的功能有关。为了克服这个问题，我开发了新的方法来创建EBV菌株，其中EBNA-LP被删除或被改造以包含定义的突变。由于EBNA-LP基因的复杂性，这是一个特殊的技术挑战。利用这些病毒，我们将能够评估EBNA-LP在B细胞转化中所起的作用，并识别由EBNA-LP控制的基因。我还将阐明ena - lp与细胞蛋白相互作用的能力，确定ena - lp的哪些部分与这些蛋白结合。通过将EBNA-LP突变体与特定宿主蛋白和转化过程中的特定缺陷相互作用的能力相关联，我将确定这些相互作用的生物学意义，并阐明哪些相互作用可能为EBV提供新的治疗靶点。参与相互作用的蛋白质界面和蛋白质修饰位点可能为抑制这些相互作用的化合物提供靶标。准确地揭示EBNA-LP对病毒的作用及其机制，将有助于未来开发新的抗EBV治疗方法，从而可能消除病毒或EBV相关癌症的新治疗方法。在癌症生物学和免疫逃避中有许多例子，通过研究已经进化到干扰细胞过程的病毒蛋白，发现了关键的细胞机制。因此，希望本研究计划中的实验也能对疾病机制产生更普遍的见解。

项目成果

Epstein-Barr virus nuclear antigen EBNA-LP is essential for transforming naive B cells, and facilitates recruitment of transcription factors to the viral genome

Epstein-Barr 病毒核抗原 EBNA-LP 对于转化幼稚 B 细胞至关重要，并有助于将转录因子招募到病毒基因组中

• DOI: 10.1101/176099
• 发表时间: 2017
• 作者: Szymula A

Heterogeneity of the Epstein-Barr Virus (EBV) Major Internal Repeat Reveals Evolutionary Mechanisms of EBV and a Functional Defect in the Prototype EBV Strain B95-8.

• DOI: 10.1128/jvi.00920-17
• 发表时间: 2017-12-01
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Ba Abdullah MM;Palermo RD;Palser AL;Grayson NE;Kellam P;Correia S;Szymula A;White RE
• 通讯作者: White RE

EBV epigenetically suppresses the B cell-to-plasma cell differentiation pathway while establishing long-term latency.

• DOI: 10.1371/journal.pbio.2001992
• 发表时间: 2017-08
• 期刊: PLoS biology
• 影响因子: 9.8
• 作者: Styles CT;Bazot Q;Parker GA;White RE;Paschos K;Allday MJ
• 通讯作者: Allday MJ

Epstein-Barr virus nuclear antigen EBNA-LP is essential for transforming naïve B cells, and facilitates recruitment of transcription factors to the viral genome.

• DOI: 10.1371/journal.ppat.1006890
• 发表时间: 2018-03
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Szymula A;Palermo RD;Bayoumy A;Groves IJ;Ba Abdullah M;Holder B;White RE
• 通讯作者: White RE

Correction: Epstein-Barr virus nuclear antigen EBNA-LP is essential for transforming naïve B cells, and facilitates recruitment of transcription factors to the viral genome.

更正：Epstein-Barr 病毒核抗原 EBNA-LP 对于转化幼稚 B 细胞至关重要，并有助于将转录因子招募到病毒基因组中。

• DOI: 10.1371/journal.ppat.1007403
• 发表时间: 2019
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Szymula A