Kedar P S, Widen S G, Englander E W, Fornace A J, Wilson S H
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1991 May 1;88(9):3729-33. doi: 10.1073/pnas.88.9.3729.
DNA polymerase beta (pol beta) is a constitutively expressed DNA repair enzyme in vertebrate cells. Yet, it had been shown previously that the pol beta mRNA level increases in Chinese hamster ovary (CHO) cells within 4 h after treatment with several monofunctional DNA damaging agents, notably, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Herein we report that a transfected pol beta promoter fusion gene is activated by MNNG treatment of CHO cells; mRNA from the transfected gene is approximately 10-fold higher in treated cells than in untreated cells 16 h after treatment. This activation is mediated through the decanucleotide palindromic element GTGACGTCAC at positions -49 to -40 in the "TATA-less" core promoter. This element, which is similar to the ATF/CREB transcription factor-binding site in a number of mammalian genes, forms the center of a strong protein-binding site for CHO cell nuclear extract proteins. Mutated pol beta promoter fusion genes lacking the element fail to bind protein at this site and fail to respond to MNNG treatment of cells.
DNA聚合酶β(polβ)是脊椎动物细胞中一种组成型表达的DNA修复酶。然而,先前已经表明,在用几种单功能DNA损伤剂处理后4小时内,中国仓鼠卵巢(CHO)细胞中的polβmRNA水平会升高,特别是N-甲基-N'-硝基-N-亚硝基胍(MNNG)。在此我们报告,转染的polβ启动子融合基因在CHO细胞经MNNG处理后被激活;处理16小时后,转染基因的mRNA在处理细胞中比未处理细胞中高约10倍。这种激活是通过“无TATA”核心启动子中-49至-40位的十核苷酸回文元件GTGACGTCAC介导的。该元件类似于许多哺乳动物基因中的ATF/CREB转录因子结合位点,形成了CHO细胞核提取物蛋白的强蛋白结合位点的中心。缺乏该元件的突变polβ启动子融合基因在该位点无法结合蛋白,并且对细胞的MNNG处理无反应。