TRANSLATIONAL CONTROL OF CYTOMEGALOVIRUS GENE EXPRESSION

巨细胞病毒(CMV)是一种人类疱疹病毒 新生儿、艾滋病患者、 移植受者和其他免疫功能受损的患者。 CMV基因表达的调控是一项复杂的任务 需要顺序和协调的表达 大约一百个基因。以下转录本 激活，转录后事件在 控制CMV基因表达的动力学和程度。这个 这项研究的主要目标是阐明翻译和 其他转录后机制，控制着 巨细胞病毒基因的表达。暂态试验和体外试验 翻译分析将用于识别新的顺式作用信号 它们影响CMV基因的表达，集中在 DNA聚合酶基因pp150和主要的2.7kbβ基因。这个 每个信号作用的特定转录后水平 将通过使用转录本的亚细胞定位来确定 杂交分析。上游AUG的调节作用 密码子，显然是CMV基因的常见元件 在几个系统中已知为平移控制信号，将 通过缺失和定点突变分析确定。 因为这些顺式作用信号很可能是 细胞或病毒反式作用因子，反式作用的作用 观察到的翻译规则中的因素将是 采用共转染法和体外翻译试验进行研究。 2.7kb的β基因编码最丰富的病毒转录本 感染早期，其基因产物(S)尚不清楚。 因为该基因在CMV生物学和 因为它含有翻译控制信号，基因产物 (S)将用多克隆抗肽抗血清进行鉴定。最后， 过去对CMV基因调控的研究主要集中在 细胞培养中的生产性感染。最终，剖析 自然发生过程中CMV基因表达的调控 感染，无论是生产性的、潜伏性的还是流产的 需要了解病毒的生物学并制定 旨在控制感染的战略。CMV调控基因 在非允许细胞中的表达将被检测以确定 翻译控制在限制寄主范围中的作用 CMV.原位杂交和抗原检测技术将 用于研究CMV基因在自然发生过程中的表达 人体组织中的感染。

Cytomegalovirus (CMV) is a human herpesvirus responsible for considerable morbidity and mortality in newborns, AIDS patients, transplant recipients and other immunocompromised patients. The regulation of CMV gene expression is a complex task requiring the sequential and coordinate expression of approximately one hundred genes. Following transcriptional activation, posttranscriptional events play a prominent role in controlling the kinetics and extent of CMV gene expression. The primary goal of this research is to elucidate translational and other posttranscriptional mechanisms which control the expression of CMV genes. Transient assays and in vitro translation assays will be used to identify new cis-acting signals which influence the expression of the CMV genes focusing on the DNA polymerase gene, pp150 and the major 2.7kb beta gene. The specific posttranscriptional level at which each signal operates will be determined by subcellular localization of transcripts using hybridization analyses. The regulatory role of upstream AUG codons, apparently common elements of CMV genes which are known to be translational control signals in several systems, will be ascertained by deletion and site specific mutation analyses. Since it is likely that these cis-acting signals are targets of cellular or viral trans-acting factors, the role of trans-acting factors in the observed translational regulation will be investigated using cotransfection and in vitro translation assays. The 2.7kb beta gene encodes the most abundant viral transcript at early times of infection yet its gene product(s) is unknown. Because of the likely importance of this gene in CMV biology and because it contains translational control signals, the gene product (s) will be identified using polyclonal antipeptide antisera. Finally, past studies of CMV gene regulation have concentrated on productive infections in cell culture. Ultimately, dissection of the regulation of CMV gene expression in naturally occurring infections whether productive, latent or abortive will be necessary to understand the biology of the virus and to formulate strategies aimed at controlling infection. CMV-regulated gene expression in nonpermissive cells will be assayed to determine the role of translational controls in limiting the host range of CMV. In situ hybridization and antigen detection techniques will be used to study CMV gene expression during naturally occurring infections in human tissues.