Jost Jean-Pierre, Thiry Stéphane, Siegmann Michel
Friedrich Miescher Institute, Maulbeerstrasse 66, CH-4058, Basel, Switzerland.
FEBS Lett. 2002 Sep 11;527(1-3):63-6. doi: 10.1016/s0014-5793(02)03166-6.
At a concentration of 5 x 10(-9) M of hemi-methylated DNA (one order of magnitude below the K(m)), MCF-7 (a human breast carcinoma cell line) nuclear extracts potentiate the activity of 5-methylcytosine DNA glycosylase (5-MCDG, alias G/T mismatch DNA glycosylase). Depending on the ratio between MCF-7 nuclear extracts and 5-MCDG, there is an up to 10-fold increase in 5-MCDG activity. The potentiation of 5-MCDG by MCF-7 nuclear extracts requires an estradiol response element adjacent to the hemi-methylated site. Depletion of the estradiol receptor from MCF-7 nuclear extracts with specific antibodies abolishes the potentiation of 5-MCDG activity. The estradiol receptor present in MCF-7 nuclear extracts can be precipitated with antibodies directed against 5-MCDG. Reciprocally, antibodies directed against the estradiol receptor precipitate 5-MCDG. The results indicate the formation of a complex between the estradiol receptor and 5-MCDG.
在半甲基化DNA浓度为5×10⁻⁹ M(比米氏常数低一个数量级)时,MCF-7(一种人乳腺癌细胞系)核提取物可增强5-甲基胞嘧啶DNA糖基化酶(5-MCDG,别名G/T错配DNA糖基化酶)的活性。根据MCF-7核提取物与5-MCDG的比例,5-MCDG活性可提高多达10倍。MCF-7核提取物对5-MCDG的增强作用需要半甲基化位点附近存在雌激素反应元件。用特异性抗体从MCF-7核提取物中去除雌激素受体可消除5-MCDG活性的增强。MCF-7核提取物中存在的雌激素受体可用针对5-MCDG的抗体沉淀。相反,针对雌激素受体的抗体可沉淀5-MCDG。结果表明雌激素受体与5-MCDG之间形成了复合物。