Contributions of Epstein-Barr Virus EBNA2 and EBNA-LP to B cell immortalization
Contributions of Epstein-Barr Virus EBNA2 and EBNA-LP to B cell immortalization
批准号:
7317804
负责人:
Chisaroka W Echendu
金额:
$2.63万
依托单位国家:
美国
项目类别:
财政年份:
2007
资助国家:
美国
项目状态:
已结题
起止时间:
2007-01-01 至 2008-07-27
关键词:
Amino AcidsAutoimmune DiseasesB lymphocyte immortalizationB-LymphocytesBindingBiological AssayCell physiologyComplement 3d ReceptorsDNA-Protein InteractionDimerizationDominant-Negative MutationGene ExpressionGenerationsGenesGeneticGenetic TranscriptionHomoHumanHuman Herpesvirus 4In VitroLMP1LengthLocalizedMediatingMethylationModificationMutationOncogene ProteinsRoleTestingTherapeuticTrans-ActivatorsTranscriptional ActivationViralViral Proteinscell transformationinhibitor/antagonistmutantpromotersmall moleculevirus related cancer
中文摘要
项目概述:EB病毒(EBV)在体外有效地永生化人类B细胞,这需要
病毒蛋白EBNA 2和EBNA-LP的表达。EBNA 2是病毒和细胞基因的反式激活因子
表情EBNA-LP是EBNA 2的基因特异性共激活因子,其上调主要的
病毒癌蛋白LMP 1。我们实验室的广泛目标是了解EBNA 2和EBNA-LP在以下方面的作用:
调节促进B细胞永生化的细胞过程。目标1.研究EBNA 2的作用
转录激活和B细胞永生化中的氨基酸残基1-58。EBNA 2氨基酸残基
1-58对全长EBNA 2具有显性负效应。EBNA 2内的特异性突变保守
对应于残基1-58的区域(CR)1和2导致缺陷性EBNA 2同源寡聚化。我们
已经获得了也不能寡聚化的功能缺陷突变体。我们将测试这些
突变体在遗传互补测定中维持B细胞永生化的能力。功能
将在EBV阳性B细胞中进行测定,以确定这些突变型EBNA 2诱导
LMP1。qRT-PCR将用于测试CR 1和/或CR 2是否介导全局或基因特异性EBNA 2
活动目标二。确定EBNA-LP介导的SplOO从PML NB置换的机制,
以及这如何促进EBNA 2共激活。SplOO氨基酸残基3-152介导二聚化,
PML NB定位和与EBNA-LP的相互作用。为了确定EBNA-LP如何重新定位SplOO,
在PML NB中,连续15个氨基酸缺失已被引入到SplOO中残基1- 10之间。
150.突变体将用于Co-IP和IF测定,以确定介导自身免疫的关键SplOO残基。
结合、EBNA-LP结合和PML NB定位。EBNA-LP、SplOO和EBNA 2的关联,
以及LMP 1启动子上的特异性修饰(例如甲基化)将通过ChIP
测定。相关性:靶向寡聚化的EBNA 2功能的小分子抑制剂可能是一种富有成效的
EBV相关癌症的治疗方法。我们将阐明EBNA-LP在
B细胞永生化,以及SplOO的正常作用,特别是与自身免疫性疾病有关。
英文摘要
Project Summary: Epstein-Barr Virus (EBV) efficiently immortalizes human B cells in vitro and this requires
expression of the viral proteins EBNA2 and EBNA-LP. EBNA2 is a transactivator of viral and cellular gene
expression. EBNA-LP is a gene-specific coactivator of EBNA2, which up-regulates expression of the major
viral oncoprotein LMP1. The broad objective of our lab is to understand the role of EBNA2 and EBNA-LP in
modulating cellular processes that promote B cell immortalization. Aim 1. Investigate the role of EBNA2
amino acid residues 1-58 in transcription activation and B cell immortalization. EBNA2 amino acid residues
1-58 have a dominant negative effect on full length EBNA2. Specific mutations within EBNA2 conserved
regions (CR) 1 and 2, corresponding to residues 1-58, result in defective EBNA2 homo-oligomerization. We
have obtained functionally-deficient mutants which are also unable to oligomerize. We will test these
mutants in genetic complementation assays for their ability to maintain B cell immortalization. Functional
assays will be performed in EBV-positive B cells to determine the ability of these mutant EBNA2 to induce
LMP1. qRT-PCR will be used to test whether CR1 and/or CR2 mediate global or gene-specific EBNA2
activity. Aim 2. Determine the mechanism of EBNA-LP-mediated displacement of SplOO from PML NBs,
and how this contributes to EBNA2 coactivation. SplOO amino acid residues 3-152 mediate dimerization,
PML NB localization, and interactions with EBNA-LP. To determine how EBNA-LP re-localizes SplOO out
of PML NBs, consecutive 15 amino acid deletions have been introduced into SplOO between residues 1-
150. The mutants will be used in Co-IP and IF assays to define critical SplOO residues that mediate self-
association, EBNA-LP binding and PML NB localization. The association of EBNA-LP, SplOO, and EBNA2,
as well as specific modifications (e.g. methylation) on the LMP1 promoter will be determined by ChIP
assays. Relevance: Small molecule inhibitors of EBNA2 function that target oligomerization may be a fruitful
therapeutic approach for EBV-related cancers. We will clarify the mechanistic contributions of EBNA-LP in
B cell immortalization, as well as the normal role of SplOO, especially as related to autoimmune diseases.
期刊论文(1)
专著(0)
科研奖励(0)
会议论文
Regulation of Sp100A subnuclear localization and transcriptional function by EBNA-LP and interferon.
DOI:
10.1089/jir.2008.0023
发表时间:
2008-10
期刊:
Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research
影响因子:
--
作者:
[Chisaroka W Echendu;P. Ling]
通讯作者:
Chisaroka W Echendu;P. Ling
Contributions of Epstein-Barr Virus EBNA2 and EBNA-LP to B cell immortalization
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批准号:7231100
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项目类别:
-
资助金额:$3.14万
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财政年份:2007
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负责人:Chisaroka W Echendu
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依托单位:
国内基金
海外基金
Autoimmune diseases therapies: variations on the microbiome in rheumatoid arthritis
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批准号:31171277
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项目类别:面上项目
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资助金额:60.0万元
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批准年份:2011
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负责人:Christine Nardini
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依托单位: