Brown S D, Van der Ploeg L H
Department of Pharmacology, Columbia University, New York, NY 10032.
Mol Biochem Parasitol. 1994 May;65(1):109-22. doi: 10.1016/0166-6851(94)90120-1.
We performed gel retardation analyses of DNA-protein interactions using DNA from the procyclic acidic repetitive protein (PARP) promoter of the protozoan parasite Trypanosoma brucei. The PARP genes of Trypanosoma brucei are transcribed in an alpha-amanitin resistant manner, and it has been proposed that RNA polymerase I, rather than RNA polymerase II, transcribes the PARP genes. Double-stranded restriction fragments containing the essential PARP-promoter regions bound only sequence-nonspecific nuclear factors, even though protein factors that bind specifically to double-stranded DNA from the snRNA U2 promoter were present in the extracts. In contrast, single-stranded DNA-binding proteins bound with high affinity, nucleotide-sequence and strand-specificity to the -69/-55 element and the coding and non-coding strands of the -37/-11 element.
我们使用原生动物寄生虫布氏锥虫的前循环酸性重复蛋白(PARP)启动子的DNA进行了DNA-蛋白质相互作用的凝胶阻滞分析。布氏锥虫的PARP基因以α-鹅膏蕈碱抗性方式转录,并且有人提出是RNA聚合酶I而非RNA聚合酶II转录PARP基因。即使提取物中存在与snRNA U2启动子的双链DNA特异性结合的蛋白质因子,但含有PARP启动子必需区域的双链限制性片段仅结合序列非特异性核因子。相比之下,单链DNA结合蛋白以高亲和力、核苷酸序列和链特异性与-69 / -55元件以及-37 / -11元件的编码链和非编码链结合。