Lee H J, Lee Y, Burbach J P, Chang C
Endocrinology-Reproductive Physiology Program, University of Wisconsin, Madison 53792, USA.
J Biol Chem. 1995 Dec 15;270(50):30129-33. doi: 10.1074/jbc.270.50.30129.
The key expression of the simian virus 40 (SV40) major late promoter could be repressed by the human TR4 orphan receptor via the +55 region of the SV40 major late promoter (nucleotide numbers 368-389, 5' -GTTA-AGGTTCGTAGGTCATGGA-3'). Using the coupled in vitro transcribed and translated TR4 orphan receptor with a molecular mass of 67.3 kilodaltons, electrophoretic mobility shift assay showed specific binding with a dissociation constant of 1.09 nM between the TR4 orphan receptor and the SV40 +55 oligonucleotides. In addition, chloramphenicol acetyltransferase assay demonstrated that this SV40 +55 region can function as a repressor via the TR4 orphan receptor, suppressing the transcriptional activities of both SV40 early and late promoters. Together, our data suggest that the TR4 orphan receptor may play an important role for the suppression of the SV40 gene expression.
猿猴病毒40(SV40)主要晚期启动子的关键表达可被人类TR4孤儿受体通过SV40主要晚期启动子的+55区域(核苷酸编号368 - 389,5'-GTTA-AGGTTCGTAGGTCATGGA-3')所抑制。使用分子量为67.3千道尔顿的体外转录和翻译偶联的TR4孤儿受体,电泳迁移率变动分析显示TR4孤儿受体与SV40 +55寡核苷酸之间存在特异性结合,解离常数为1.09 nM。此外,氯霉素乙酰转移酶分析表明,该SV-40 +55区域可通过TR4孤儿受体发挥阻遏物的作用,抑制SV40早期和晚期启动子的转录活性。总之,我们的数据表明TR4孤儿受体可能在抑制SV40基因表达中发挥重要作用。