Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells.
Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells.
复制标题
转基因诱导的 CCWGG 甲基化不会改变人肾细胞中的 CG 甲基化模式。
DOI：
10.1093/nar/gki920
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发表时间：
2005
影响因子：
14.9
通讯作者：
Smith,StevenS
中科院分区：
文献类型：
--
作者：
Shevchuk,Taras;Kretzner,Leo;Munson,Kristofer;Axume,John;Clark,Jarrod;Dyachenko,OlgaV;Caudill,Marie;Buryanov,Yaroslav;Smith,StevenS
一些报告表明，在人类细胞中，CmCWGG甲基化倾向于不与mCG甲基化共存。我们已经询问了CCWGG位点的甲基化是否会影响CG甲基化。来自表达M.EcoRII-GFP融合体的细胞的DNA在CCWGG位点被主动甲基化。在表达转基因的细胞中，通过R.HpaII/R.MspI比率测量的CG甲基化没有变化。CmCWGG甲基化DNA的克隆代表通常包含或邻近ALU重复，表明M.EcoRII-GFP主动甲基化富含基因的R带DNA。转基因甲基转移酶将CmCWGG甲基化应用于CG二核苷酸高度甲基化的代表性人类启动子（SERPINB 5启动子）和CG二核苷酸基本未甲基化的代表性启动子（APC启动子）。在每种情况下，CG甲基化模式保持在其原始状态，未因相邻CmCWGG位点的存在而改变。Q-PCR测量结果表明，APC基因的RNA表达没有显着改变CmCWGG在其启动子中的存在。动力学研究表明，相邻的CmCWGG甲基化位点既不影响纯化的人Dnmt 1的维持，也不影响其去新甲基化活性。我们的结论是，CmCWGG甲基化不发挥CG甲基化在人肾细胞中的显着影响。
Several reports suggest that CmCWGG methylation tends not to co-exist withmCG methylation in human cells. We have asked whether or not methylation at CCWGG sites can influence CG methylation. DNA from cells expressing an M.EcoRII–GFP fusion was actively methylated at CCWGG sites. CG methylation as measured by R.HpaII/R.MspI ratios was unchanged in cells expressing the transgene. Cloned representatives of CmCWGG methylated DNA often contained, or were adjacent to an ALU repeat, suggesting that M.EcoRII-GFP actively methylated gene-rich R-band DNA. The transgenic methyltransferase applied CmCWGG methylation to a representative human promoter that was heavily methylated at CG dinucleotides (the SERPINB5 promoter) and to a representative promoter that was essentially unmethylated at CG dinucleotides (the APC promoter). In each case, the CG methylation pattern remained in its original state, unchanged by the presence of neighboring CmCWGG sites. Q-PCR measurements showed that RNA expression from the APC gene was not significantly altered by the presence of CmCWGG in its promoter. Kinetic studies suggested that an adjacent CmCWGG methylation site influences neither the maintenance nor thede novomethylation activities of purified human Dnmt1. We conclude that CmCWGG methylation does not exert a significant effect on CG methylation in human kidney cells.