HCMV UL78 expression and function

HCMV UL78 表达和功能

• 批准号: 6694819
• 负责人: DAVID M BROWN
• 金额: $ 3.97万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6694819
• 关键词: artificial chromosomes; cell line; chemokine; cytomegalovirus; enzyme linked immunosorbent assay; epitope mapping; gene expression; laboratory mouse; microarray technology; mutant; northern blottings; postdoctoral investigator; protein localization; protein structure function; virus genetics; virus protein; western blottings

DESCRIPTION (provided by applicant): Human Cytomegalovirus (HCMV) causes morbidity and mortality in neonates and in immunocompromised individuals. HCMV encodes at least four genes (UL33, UL78, US27, and US28) with homology to G-protein coupled receptors (GPCRs). Studies of M78, the murine homologue of UL78, have shown that M78 is an integral protein in the envelope of virions. Mutations in this gene result in a viral growth defect and reduced expression of an immediate early gene, m123. Interestingly, the de novo expression of M78 is not required to mediate this effect suggesting that the M78 protein present in the virion envelope is sufficient for the efficient expression of m123. This research proposal will: 1.) Determine the expression pattern of UL78 mRNA and protein, 2.) Determine the growth properties of a UL78 deleted mutant virus and 3.) Determine whether the presence of UL78 protein by itself, or in the presence of a CC chemokine (RANTES, MCP-1, MIP-1alpha and MIP-1beta), influences the expression of HCMV and/or cellular genes. Protein expression and localization will be determined by Western blot analysis and/or epitope tagging of the C-terminal domain of UL78. Mutant virus will be generated in an infectious HCMV bacterial artificial chromosome (BAC) and, if necessary, utilizing a complementing cell-line. Growth properties will be determined at a high and low multiplicity of infection in several primary or life-extended cell types. Effects on HCMV or cellular gene expression patterns can be monitored using a viral gene array or cellular affymetrix array respectively, and confirmed by Northern blot analysis.

描述（由申请人提供）：人类巨细胞病毒（HCMV）导致新生儿和免疫功能低下的个体的发病率和死亡率。 HCMV用与G蛋白偶联受体（GPCRS）同源的至少四个基因（UL33，UL78，US27和US28）。 M78的研究是UL78的鼠同源物，它表明M78是病毒体包膜中的积分蛋白。 该基因中的突变导致病毒生长缺陷并降低了早期基因M123的表达。 有趣的是，不需要M78的从头表达来介导这种作用，这表明存在于病毒素包膜中的M78蛋白足以有效地表达M123。 This research proposal will: 1.) Determine the expression pattern of UL78 mRNA and protein, 2.) Determine the growth properties of a UL78 deleted mutant virus and 3.) Determine whether the presence of UL78 protein by itself, or in the presence of a CC chemokine (RANTES, MCP-1, MIP-1alpha and MIP-1beta), influences the expression of HCMV and/or cellular genes.蛋白质的表达和定位将通过UL78的C末端结构域的蛋白质印迹分析和/或表位标记来确定。突变病毒将在传染性的HCMV细菌人造染色体（BAC）中产生，并在必要时使用互补的细胞线。 在几种原发性或延伸的细胞类型中，将在高和低的感染中确定生长特性。 可以分别使用病毒基因阵列或细胞伴侣阵列来监测对HCMV或细胞基因表达模式的影响，并通过Northern blot分析确认。