IDENTIFICATION AND CHARACTERIZATION OF HUMAN CYTOMEGALOVIRUS/CELLULAR PROTEINS

人类巨细胞病毒/细胞蛋白的鉴定和表征

• 批准号: 7667995
• 负责人: Y JERVEY TABMITHA
• 金额: $ 14.31万
• 依托单位: NORFOLK STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7667995
• 关键词: Address; Adult; Antiviral Therapy; Benign; Binding; Binding Proteins; Binding Sites; Biological Assay; Cells; Class; Cyclic AMP-Responsive DNA-Binding Protein; Cytomegalovirus; Cytomegalovirus Infections; DNA; DNA Probes; DNA Sequence; DNA biosynthesis; Development; Early Promoters; Electrophoretic Mobility Shift Assay; Elements; Exonuclease; Family; Fetus; Future; Gel; Gene Expression; Gene Expression Regulation; Genes; Genome; Goals; Homologous Gene; Immediate-Early Genes; Immune; Immunocompromised Host; Individual; Infant; Infection; Interferon Type II; Nuclear Extract; Open Reading Frames; Participant; Pathogenesis; Phase; Phosphoproteins; Play; Polyadenylation; Polymerase Gene; Process; Production; Protein Binding; Proteins; RNA; Regulation; Response Elements; Role; Simplexvirus; Site; T-Lymphocyte; Time; Trans-Activators; Transcript; Transfection; Viral; activating transcription factor; base; cis acting element; common cellular transcription factor ATF; deletion analysis; expectation; gel mobility shift assay; in utero; insight; interest; latent infection; novel; pathogen; promoter; superinfection; transcription factor

Human cytomegalovirus (HCMV) is a widespread opportunistic pathogen capable of establishing either a persistent or latent infection. In most cases, HCMV infection is benign or asymptomatic in healthy ndividuals. However, serious illness occurs upon infection in infants and immunocompromised adults. The genome is expressed in et temporally ordered manner and occurs in three separate phases: immediate early IE), early, and late. The early phase, which begins just prior to DNA replication, is further divided into three subclasses based on the expression level of early transcripts through the course of infection. / Studies on several HCMV early promoters have been undertaken to determine how HCMV early gene expression is regulated. Several specific DNA sequences as well as proteins have been identified. Some HCMV early promoter sequences are bound by HCMV specific proteins (IE86) as well as such as cellular proteins such as the cyclic AMP response element binding protein (CREB) or activated transcription factor [ATF). Activation of the early promoters requires the presence of IE86 and IE72 proteins or IE86 alone. * The UL98 early gene encodes a 58-65 kDa protein and is part of a family of nested 3' coterminal ORFs that share a polyadenylation site. The alkaline exonuclease plays a critical role in processing newly synthesized replicated copies of genome during maturation. We have determined that the HCMV UL98 promoter utilizes CRE and gamma (g) IRE to optimally regulate the expression of this early gene. In deletion analysis, a Tcell factor-like element (TCP) has also been implicated. Studies have also shown that the protein CREB interacts with the CRE sequences in gel mobility shift assays. However, the specific proteins associated with regulation through the TCF-1 and gIRE sites have yet to be characterized. Our objective in the following studies will be to identify the viral/cellular protein(s) that bind the TCF-1 element and gIRE. The analysis of regulatory mechanisms for this key viral early gene will provide insight into the overall regulation of early gene expression and the future development of novel antiviral therapies.

人巨细胞病毒 (HCMV) 是一种广泛传播的机会性病原体，能够建立 持续或潜伏感染。在大多数情况下，HCMV 感染在健康人中是良性的或无症状的 个人。然而，婴儿和免疫功能低下的成年人感染后会出现严重疾病。这 基因组以时间顺序的方式表达，并发生在三个不同的阶段：立即早期 IE），早，晚。早期阶段在 DNA 复制之前开始，进一步分为三个阶段 基于感染过程中早期转录物的表达水平的亚类。 / 已经对几个 HCMV 早期启动子进行了研究，以确定 HCMV 早期基因如何 表达受到调节。一些特定的 DNA 序列和蛋白质已被鉴定。一些 HCMV 早期启动子序列与 HCMV 特异性蛋白 (IE86) 以及细胞蛋白结合 蛋白质，例如环 AMP 反应元件结合蛋白 (CREB) 或激活的转录因子 [自动变速箱油]。早期启动子的激活需要 IE86 和 IE72 蛋白或单独的 IE86 的存在。 * UL98 早期基因编码 58-65 kDa 蛋白质，属于嵌套 3' 共末端 ORF 家族的一部分， 共享一个聚腺苷酸化位点。碱性核酸外切酶在加工新合成的过程中起着至关重要的作用 成熟过程中基因组的复制副本。我们已经确定 HCMV UL98 启动子利用 CRE 和 gamma (g) IRE 可优化调节该早期基因的表达。在缺失分析中，T细胞 类因子元素（TCP）也受到牵连。研究还表明，CREB蛋白 在凝胶迁移率变化测定中与 CRE 序列相互作用。然而，相关的特定蛋白质 通过 TCF-1 和 gIRE 位点的调节尚未得到表征。我们的目标如下 研究将鉴定结合 TCF-1 元件和 gIRE 的病毒/细胞蛋白。的分析 这一关键病毒早期基因的调控机制将有助于深入了解早期病毒的整体调控 基因表达和新型抗病毒疗法的未来发展。