Effect of potassium on DNA methylation of aldosterone synthase gene
Effect of potassium on DNA methylation of aldosterone synthase gene
复制标题
钾对醛固酮合酶基因DNA甲基化的影响
DOI：
10.1097/hjh.0000000000002742
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发表时间：
2021
影响因子：
4.9
通讯作者：
Takeda Yoshiyu
中科院分区：
文献类型：
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作者：
Takeda Yoshimichi;Demura Masashi;Wang Fen;Karashima Shigehiro;Yoneda Takashi;Kometani Mitsuhiro;Aomo Daisuke;Hashimoto Atsushi;Horike Shin-ichi;Meguro-Horike Makiko;Takeda Yoshiyu
背景：醛固酮合成酶基因CYP 11B 2受钾离子和血管紧张素II（Ang II）的调节。我们已经报道了Ang II可以改变转录因子结合位点和转录起始位点（TSS）周围的DNA甲基化状态，并激活CYP 11B 2的表达。类似于血管紧张素II，细胞外钾水平的小幅增加也增加CYP 11B 2 mRNA的水平。方法和结果：肾上腺皮质H295 R细胞用不同剂量的钾。用亚硫酸氢盐测序法对CYP 11B 2启动子区进行甲基化分析。CYP 11B 2 mRNA和蛋白水平，染色质可及性，甲基化和去甲基化活性进行了估计。检测CYP 11B 2基因启动子甲基化前后的转录能力。钾刺激引起DNA去甲基化周围的环磷酸腺苷反应元件结合蛋白1（CREB 1）和核受体亚家族4组A（NR 4A）家族结合位点和TSS;去甲基化伴随着CREB 1和NR 4A 1的招聘和增加染色质的可及性CYP 11B 2启动子。细胞核DNA甲基化活性降低。在钾（16 mmol/l）刺激后2至4天内检测到CpG 1（Ad 1）、CpG 2（Ad 5）和CpG 3的DNA去甲基化。这些变化在第7天达到最高水平。CpG 2（Ad 5）和CpG 3处的DNA在第9天重新甲基化至与未处理细胞相似的水平。钾处理显着降低DNA甲基化活性在第7和第9天。结论：钾离子可可逆地诱导DNA去甲基化，使CYP 11B 2表达表型从失活状态转变为活性状态。
Background:Aldosterone synthase gene, CYP11B2 is regulated by potassium and angiotensin II (Ang II). We have reported that Ang II could change the DNA methylation status around transcription factor-binding sites and a transcription start site (TSS) and activate expression of CYP11B2. Similar to Ang II, small increases in extracellular potassium levels also increase CYP11B2 mRNA levels.Methods and results:Adrenocortical H295R cells were treated with different doses of potassium. Methylation analysis of CYP11B2 promoter region was done by bisulfite sequencing. CYP11B2 mRNA and protein levels, chromatin accessibility, methylation and demethylation activity were estimated. The transcriptional ability of CYP11B2 promoter with or without methylation was assessed. Potassium stimulation caused DNA demethylation around cyclic AMP responsive element binding protein 1 (CREB1) and nuclear receptor subfamily 4 group A (NR4A) family-binding sites and a TSS; demethylation was accompanied by recruitment of CREB1 and NR4A1 and increased chromatin accessibility of the CYP11B2 promoter. DNA methylation activity decreased in the nucleus. DNA demethylation at CpG1 (Ad1), CpG2 (Ad5) and CpG3 were detected within 2 to 4 days after potassium (16 mmol/l) stimulation. The changes reached a maximum level by day 7. DNA at CpG2 (Ad5) and CpG3 was re-methylated to levels that were similar to those of nontreated cells at day 9. Potassium treatment significantly reduced DNA methylation activity at days 7 and 9. DNA demethylation activity was not changed by potassium.Conclusion: Potassium induced reversibly DNA demethylation, which switches the phenotype of CYP11B2 expression from an inactive to an active state.