Subramaniam M, Harris S A, Rasmussen K, Spelsberg T C
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905.
Endocrinology. 1993 Nov;133(5):2049-54. doi: 10.1210/endo.133.5.8404652.
Previous work in this and other laboratories has shown that steroids rapidly regulate the expression of nuclear protooncogenes. In this present study, we have investigated the effect of progesterone (Pg) on the expression of c-jun in the avian oviduct system and its promoter activity in avian liver cells. Pg treatment of estrogen-withdrawn chickens brings about a decrease in the steady state mRNA level of the protooncogene c-jun within 30 min. This decrease is steroid dose dependent and gene specific. Using nuclear run-off transcription analyses, this rapid regulation was shown to occur at the level of gene transcription, as the rate of c-jun transcription decreases by more than 80% within 15 min after progesterone treatment. As expected, ovalbumin gene transcription is increased only after a lag period of 4 h following Pg treatment. In other studies, we have linked the c-jun promoter sequences between -1000 and +192 to chloramphenicol acetyltransferase reporter gene and cotransfected them into transformed avian liver cells along with the expression vector for the Pg receptor. Pg treatment of these cells causes a decrease in chloramphenicol acetyltransferase gene expression, albeit to a lesser extent than Pg inhibition of c-jun gene transcription. These results suggest that the 5'-domain of the chicken c-jun gene contains sequence elements that negatively regulate c-jun promoter activity in response to Pg.
本实验室及其他实验室之前的研究表明,类固醇可快速调节核原癌基因的表达。在本研究中,我们调查了孕酮(Pg)对禽输卵管系统中c-jun表达及其在禽肝细胞中启动子活性的影响。用Pg处理摘除雌激素的鸡,30分钟内原癌基因c-jun的稳态mRNA水平就会下降。这种下降具有类固醇剂量依赖性和基因特异性。通过核转录分析表明,这种快速调节发生在基因转录水平,因为在孕酮处理后15分钟内,c-jun的转录速率下降了80%以上。正如预期的那样,卵清蛋白基因转录仅在Pg处理4小时的延迟期后增加。在其他研究中,我们将-1000至+192之间的c-jun启动子序列与氯霉素乙酰转移酶报告基因相连,并将它们与Pg受体的表达载体一起共转染到转化的禽肝细胞中。用Pg处理这些细胞会导致氯霉素乙酰转移酶基因表达下降,尽管其程度小于Pg对c-jun基因转录的抑制。这些结果表明,鸡c-jun基因的5'结构域含有响应Pg而对c-jun启动子活性进行负调控的序列元件。