Stinski M F, Roehr T J
J Virol. 1985 Aug;55(2):431-41. doi: 10.1128/JVI.55.2.431-441.1985.
Upstream of the major immediate early gene of human cytomegalovirus (Towne) is a strong promoter-regulatory region that promotes the synthesis of 1.95-kilobase mRNA (D. R. Thomsen, R. M. Stenberg, W. F. Goins, and M. F. Stinski, Proc. Natl. Acad. Sci. U.S.A. 81:659-663, 1984; M. F. Stinski, D. R. Thomsen, R. M. Stenberg, and L. C. Goldstein, J. Virol. 46:1-14, 1983). The wild-type promoter-regulatory region as well as deletions within this region were ligated upstream of the thymidine kinase, chloramphenicol acetyltransferase, or ovalbumin genes. These gene chimeras were constructed to investigate the role of the regulatory sequences in enhancing downstream expression. The regulatory region extends to approximately 465 nucleotides upstream of the cap site for the initiation of transcription. The extent and type of regulatory sequences upstream of the promoter influences the level of in vitro transcription as well as the amount of in vivo expression of the downstream gene. The regulatory elements for cis-activation appear to be repeated several times within the regulatory region. A direct correlation was established between the distribution of the 19 (5' CCCCAGTTGACGTCAATGGG 3')- and 18 (5' CACTAACGGGACTTTCCAA 3')-nucleotide repeats and the level of downstream expression. In contrast, the 16 (5' CTTGGCAGTACATCAA 3')-nucleotide repeat is not necessary for the enhancement of downstream expression. In a domain associated with the 19- or 18-nucleotide repeats are elements that can be activated in trans by a human cytomegalovirus-specified component but not a herpes simplex virus-specified component. Therefore, the regulatory sequences of the major immediate early gene of human cytomegalovirus have an important role in interacting with cellular and virus-specific factors of the transcription complex to enhance downstream expression of this critical viral gene.
人类巨细胞病毒（汤氏株）主要立即早期基因的上游是一个强大的启动子调控区域，它能促进1.95千碱基mRNA的合成（D. R. 汤姆森、R. M. 斯滕伯格、W. F. 戈因斯和M. F. 斯廷斯基，《美国国家科学院院刊》81:659 - 663, 1984；M. F. 斯廷斯基、D. R. 汤姆森、R. M. 斯滕伯格和L. C. 戈尔茨坦，《病毒学杂志》46:1 - 14, 1983）。野生型启动子调控区域以及该区域内的缺失片段被连接到胸苷激酶、氯霉素乙酰转移酶或卵清蛋白基因的上游。构建这些基因嵌合体是为了研究调控序列在增强下游基因表达中的作用。调控区域延伸至转录起始位点帽位点上游约465个核苷酸处。启动子上游调控序列的范围和类型会影响体外转录水平以及下游基因在体内的表达量。顺式激活的调控元件似乎在调控区域内重复出现了几次。在19个（5' CCCCAGTTGACGTCAATGGG 3'）和18个（5' CACTAACGGGACTTTCCAA 3'）核苷酸重复序列的分布与下游表达水平之间建立了直接关联。相比之下，16个（5' CTTGGCAGTACATCAA 3'）核苷酸重复序列对于增强下游表达并非必需。在与19或18个核苷酸重复序列相关的结构域中存在一些元件，它们可被人类巨细胞病毒指定的成分反式激活，但不能被单纯疱疹病毒指定的成分激活。因此，人类巨细胞病毒主要立即早期基因的调控序列在与转录复合物的细胞和病毒特异性因子相互作用以增强这个关键病毒基因的下游表达方面具有重要作用。
