Wagner S, Knippers R
Fakultät für Biologie, Universität Konstanz, Federal Republic of Germany.
Oncogene. 1990 Mar;5(3):353-9.
A genomic mouse DNA fragment (Q300), containing a high affinity binding site for SV40 large T antigen, serves as a cis-acting transcriptional element in in vivo and in in vitro studies. We have performed experiments to investigate whether bound T antigen could modulate the promoter-enhancer activity of the Q300 element. In vivo studies showed a negative effect of T antigen on the Q300 driven expression of the chloramphenicol acetyl transferase reporter gene. Band shift and DNAase I protection experiments demonstrated that T antigen and a nuclear protein, probably a CCAAT-binding factor, can simultaneously bind to closely adjacent sites on the Q300 DNA. Transcription studies in vitro showed that bound T antigen suppresses the transcriptional enhancer effect of the Q300 element. We interpret the results of these model studies to indicate that T antigen, bound to DNA, is able to affect the function of a cellular cis-acting transcriptional element. Bound T antigen may influence the activity of a cellular transcription factor at a closely adjacent DNA site.
一个含有SV40大T抗原高亲和力结合位点的基因组小鼠DNA片段(Q300),在体内和体外研究中作为顺式作用转录元件。我们进行了实验,以研究结合的T抗原是否能调节Q300元件的启动子增强子活性。体内研究表明,T抗原对氯霉素乙酰转移酶报告基因的Q300驱动表达有负面影响。凝胶迁移和DNA酶I保护实验表明,T抗原和一种核蛋白(可能是CCAAT结合因子)能同时结合到Q300 DNA上紧密相邻的位点。体外转录研究表明,结合的T抗原抑制Q300元件的转录增强子效应。我们对这些模型研究结果的解释是,与DNA结合的T抗原能够影响细胞顺式作用转录元件的功能。结合的T抗原可能会影响紧密相邻DNA位点上细胞转录因子的活性。