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低下的患者。 CMV基因表达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。 转录后 激活，转录后事件发挥突出作用， 控制CMV基因表达的动力学和程度。 的 本研究的主要目标是阐明翻译和 其他转录后机制，控制 CMV基因的表达。 瞬时测定和体外 翻译分析将用于鉴定新的顺式作用信号 影响CMV基因的表达，集中在 DNA聚合酶基因、pp150和主要的2.7kb β基因。 的 每个信号在特定转录后水平起作用 将通过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基因的共同元件， 已知在几个系统中是平移控制信号， 通过缺失和位点特异性突变分析确定。 因为这些顺式作用信号很可能是 细胞或病毒的反式作用因子，反式作用的作用 观察到的翻译调节中的因素将是 使用共转染和体外翻译测定研究。 2.7kb的β基因编码最丰富的病毒转录本， 早期感染，但其基因产物是未知的。 由于该基因在CMV生物学中的可能重要性， 因为它含有翻译控制信号， (s)将使用多克隆抗肽抗血清鉴定。最后， 过去对CMV基因调控的研究主要集中在 在细胞培养中产生感染。 最终，解剖 CMV基因表达调控 感染，无论是生产性的，潜伏的还是流产的， 了解病毒的生物学特性， 旨在控制感染的战略。 CMV调控基因 将测定非允许细胞中的表达以确定 翻译控制在限制宿主范围中的作用 巨细胞病毒 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， 感染人体组织。

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.