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目的 探讨谷胱甘肽 S-转移酶 (GST- P)基因表达调控机制的多样性及其与化学致癌的关系 ,筛选与大鼠 GST- P增强子元件 GPE 相互作用的调控因子。方法 采用酵母单杂交体系筛选与 GPE 核心序列相互作用的转录激活因子 ,应用 DNA序列测定及计算机分析等手段对所测的 DNA序列进行分析。结果 共获得两个阳性克隆 p YGPE1和 p YGPE2。 DNA测序分析表明 :p YGPE1的插入片段与大鼠原癌基因 c- jun c DNA具有 99%的同源性 ,其编码的氨基酸序列与大鼠 c- Jun蛋白具有 10 0 %的同源性。 p YGPE2的插入片段与大鼠线粒体腺苷酸转位酶 c DNA具有 99%的同源性 ,其编码的氨基酸序列与大鼠腺苷酸转位酶具有 10 0 %的同源性。结论 大鼠 c- Jun蛋白和线粒体腺苷酸转位酶在酵母细胞内与大鼠 GST- P增强子 GPE 核心序列结合 ,可能是作用于 GPE 的反式作用因子。
Objective To investigate the diversity of glutathione S-transferase (GST-P) gene expression regulation mechanism and its relationship with chemical carcinogenesis, and to screen the regulatory factors that interact with GST-P enhancer element GPE in rats. Methods Yeast single-hybrid system was used to screen the transcriptional activators interacting with GPE core sequences. The DNA sequences were analyzed by DNA sequencing and computer analysis. Results A total of two positive clones p YGPE1 and p YGPE2 were obtained. DNA sequencing analysis showed that the insert of pYGPE1 is 99% homologous to the rat c-jun c DNA, and its encoded amino acid sequence is 100% homologous to rat c-Jun protein. The insert of pYGPE2 is 99% homologous to rat mitochondrial adenylate transposase c DNA, and its encoded amino acid sequence is 10% homologous to rat adenylate transposase. Conclusion The binding of c-Jun protein and mitochondrial adenylate transaminase in rat cells to the GPE core sequence of rat GST-P enhancer may be a trans-acting factor acting on GPE.