Effective, homogeneous and transient interference with cytosine methylation in plant genomic DNA by zebularine.
Effective, homogeneous and transient interference with cytosine methylation in plant genomic DNA by zebularine.
复制标题
DOI：
10.1111/j.1365-313x.2008.03699.x
复制
发表时间：
2009-02
期刊：
影响因子：
--
通讯作者：
Mittelsten Scheid O
中科院分区：
文献类型：
--
作者：
Baubec T;Pecinka A;Rozhon W;Mittelsten Scheid O
胞嘧啶残基甲基化的共价修饰是一个重要的表观遗传学特征。虽然亚硫酸氢盐转化后的序列分析允许单碱基分辨率的相关分析，但由于甲基化酶及其靶标的复杂性，通过干扰DNA甲基化进行功能分析不那么精确。胞苷类似物5-氮杂胞苷经常被用作DNA甲基转移酶的抑制剂，但它在水溶液中的快速降解在超过几个小时的培养时间内是有问题的。ZeBularine是一种更稳定的胞苷类似物，具有类似的作用模式，已成功地作为甲基化抑制剂用于脉孢子瘤和哺乳动物肿瘤细胞系，应用ZeBularine可以显著减少植物DNA甲基化，其作用方式与序列背景无关，且具有剂量依赖性和暂时性。去甲基化与异染色质的转录重新激活和部分去缩合有关。ZeBularine是一种很有前途的新的通用工具，用于研究DNA甲基化在植物转录控制、(异源)染色质的维持和形成方面的作用。
Covalent modification by methylation of cytosine residues represents an important epigenetic hallmark. While sequence analysis after bisulphite conversion allows correlative analyses with single-base resolution, functional analysis by interference with DNA methylation is less precise, due to the complexity of methylation enzymes and their targets. A cytidine analogue, 5-azacytidine, is frequently used as an inhibitor of DNA methyltransferases, but its rapid degradation in aqueous solution is problematic for culture periods of longer than a few hours. Application of zebularine, a more stable cytidine analogue with a similar mode of action that is successfully used as a methylation inhibitor in Neurospora and mammalian tumour cell lines, can significantly reduce DNA methylation in plants in a dose-dependent and transient manner independent of sequence context. Demethylation is connected with transcriptional reactivation and partial decondensation of heterochromatin. Zebularine represents a promising new and versatile tool for investigating the role of DNA methylation in plants with regard to transcriptional control, maintenance and formation of (hetero-) chromatin.