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Re-specification of the Notch response by the HHV-8 lytic switch protein

HHV-8 裂解开关蛋白对 Notch 反应的重新特异性

基本信息

批准号：
8704637
负责人：
DAVID M LUKAC
金额：
$ 20.43万
依托单位：
RBHS-NEW JERSEY MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-02-15 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8704637
关键词：
Affinity Chromatography Amino Acids Animal Viruses Architecture Binding Binding Proteins Binding Sites Biochemical Biological Biological Assay Biological Models Cells ChIP-on-chip Complex DNA DNA Binding DNA Sequence DNA Viruses DNA-Binding Proteins DNA-Protein Interaction Data Diagnostic Disease Drug Targeting EMSA Elements Enhancers Gene Expression Profile Gene Targeting Genes Genome Goals Human Human Herpesvirus 4 Human Herpesvirus 8 Human Pathology Hybridization Array Hybrids In Vitro Infection Light Lytic Mass Spectrum Analysis Modeling Molecular Mutation Notch Signaling Pathway Oligonucleotide Microarrays Pathway interactions Protein Binding Proteins Regulation Reporter Repressor Proteins Role Scientific Advances and Accomplishments Series Signal Pathway Signal Transduction Signal Transduction Pathway Specific qualifier value Transactivation Transcriptional Activation Viral Viral Proteins Virus Yeasts base cell growth chromatin immunoprecipitation genome-wide in vivo mutant notch protein novel promoter research study response screening tissue culture transcription factor virus host interaction

项目摘要

DESCRIPTION (provided by applicant): Defining the molecular interactions between a virus and its host that regulate gene-specific transactivation has been essential to understanding DNA virus persistence and replication. The Human herpesvirus-8 (HHV-8) ORF50/Rta protein is necessary and sufficient for the virus to emerge from latency and replicate (lytic reactivation). Rta interacts directly with the cellular protein called RBP-Jk, which is also required for lytic reactivation. RBP-Jk normally specifies the genes that will be activated by the cellular Notch signal transduction pathway by binding sequence specifically to DNA. In this fashion, RBP-Jk serves as a "landing pad" for the activated Notch receptor (Notch intracellular domain (NICD)). HHV-8 Rta promotes DNA binding of RBP-Jk during viral reactivation, a mechanism that is fundamentally different from that established for other RBP-Jk-activating proteins. Our preliminary data suggest a gene-specific mechanism for controlling RBP-Jk-dependent activity in HHV-8 infected cells. The overall goal of this application is to define the basic molecular mechanisms that regulate RBP-Jk dependent-transactivation in HHV-8 infected cells. Our studies will shed light on the fundamental regulation of productive and non-productive virus reactivation as determined by promoter-specific transactivation. We will therefore comprehensively identify proteins that bind to essential HHV-8 promoters (Aim 1). We will define the molecular interactions required for Rta stimulation of RBP-Jk DNA binding (Aim 2). We will determine the molecular requirements for forming functional RBP-Jk-containing promoter complexes and HHV-8 reactivation (Aim 3). A series of biochemical and molecular biological approaches are proposed. A biochemical screen for promoter-specific protein-DNA interactions is a major approach. Promoter-reporter, protein-protein and protein-DNA interactions represent the basis for many of the experiments, and include novel, highly quantitative in vitro and in vivo assays. This proposal will shed light on how Notch target genes are specified, and reveal new components of the Notch signal transduction pathway.

描述（由申请人提供）：定义病毒与其宿主之间调节基因特异性转激活的分子相互作用对于理解DNA病毒的持久性和复制至关重要。人疱疹病毒-8 (HHV-8) ORF50/Rta蛋白是病毒从潜伏期出现和复制（裂解再激活）所必需和充分的。Rta直接与称为RBP-Jk的细胞蛋白相互作用，RBP-Jk也是裂解再激活所必需的。RBP-Jk通常通过与DNA特异性结合序列指定将被细胞Notch信号转导通路激活的基因。在这种方式下，RBP-Jk作为激活的Notch受体（Notch胞内结构域（NICD））的“着陆点”。HHV-8 Rta在病毒再激活过程中促进RBP-Jk的DNA结合，这一机制与其他RBP-Jk激活蛋白的机制根本不同。我们的初步数据表明，在HHV-8感染细胞中存在一种基因特异性机制来控制rbp - jk依赖性活性。本应用程序的总体目标是确定在HHV-8感染细胞中调节RBP-Jk依赖性转激活的基本分子机制。我们的研究将阐明由启动子特异性转激活决定的生产性和非生产性病毒再激活的基本调控。因此，我们将全面鉴定与HHV-8基本启动子结合的蛋白（Aim 1）。我们将定义Rta刺激RBP-Jk DNA结合所需的分子相互作用（目的2）。我们将确定形成功能性含rbp - jk启动子复合物和HHV-8再激活的分子要求（目的3）。提出了一系列的生化和分子生物学方法。启动子特异性蛋白质- dna相互作用的生化筛选是一种主要方法。启动子-报告子，蛋白质-蛋白质和蛋白质- dna相互作用是许多实验的基础，包括新颖的，高度定量的体外和体内分析。这一建议将阐明Notch靶基因是如何被指定的，并揭示Notch信号转导途径的新成分。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

KSHV Rta Promoter Specification and Viral Reactivation.

DOI：
10.3389/fmicb.2012.00030
发表时间：
2012
期刊：
Frontiers in microbiology
影响因子：
5.2
作者：
Guito J;Lukac DM
通讯作者：
Lukac DM

Convergence of Kaposi's sarcoma-associated herpesvirus reactivation with Epstein-Barr virus latency and cellular growth mediated by the notch signaling pathway in coinfected cells.

卡波西肉瘤相关疱疹病毒再激活与 Epstein-Barr 病毒潜伏期和共感染细胞中 Notch 信号通路介导的细胞生长的收敛。

DOI：
10.1128/jvi.00894-10
发表时间：
2010
期刊：
Journal of virology
影响因子：
5.4
作者：
Spadavecchia,Sophia;Gonzalez-Lopez,Olga;Carroll,KylaDriscoll;Palmeri,Diana;Lukac,DavidM
通讯作者：
Lukac,DavidM

A herpesvirus transactivator and cellular POU proteins extensively regulate DNA binding of the host Notch signaling protein RBP-Jκ to the virus genome.

疱疹病毒反式激活蛋白和细胞 POU 蛋白广泛调节宿主 Notch 信号蛋白 RBP-Jδ 与病毒基因组的 DNA 结合。

DOI：
10.1074/jbc.ra118.007331
发表时间：
2019
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Gonzalez-Lopez,Olga;DeCotiis,Jennifer;Goyeneche,Corey;Mello,Helena;Vicente-Ortiz,BryanAlexis;Shin,HyeJin;Driscoll,KylaE;Du,Peicheng;Palmeri,Diana;Lukac,DavidM
通讯作者：
Lukac,DavidM

An easily transfectable cell line that produces an infectious reporter virus for routine and robust quantitation of Kaposi's sarcoma-associated herpesvirus reactivation.

一种易于转染的细胞系，可产生感染性报告病毒，用于卡波西肉瘤相关疱疹病毒再激活的常规和稳健定量。