Chromatin structure and methylation of rat rRNA genes studied by formaldehyde fixation and psoralen cross-linking.
Chromatin structure and methylation of rat rRNA genes studied by formaldehyde fixation and psoralen cross-linking.
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通过甲醛固定和补骨脂素交联研究大鼠 rRNA 基因的染色质结构和甲基化。
DOI：
10.1093/nar/25.9.1727
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发表时间：
1997
影响因子：
14.9
通讯作者：
J. Sogo
中科院分区：
文献类型：
--
作者：
R. Lucchini;Theodor Ko ller;J. Sogo
通过使用组蛋白与DNA的甲醛交联和凝胶延迟试验，我们表明甲醛固定，类似于先前建立的补骨脂素光交联，区分核糖体RNA基因的核小体填充(非活性)和非核小体(活性)部分。通过这两种交联技术，我们能够从琼脂糖凝胶中纯化与rRNA基因的编码序列、增强子序列和启动子序列相对应的片段，并从DNA甲基化方面进行了进一步的研究。这种方法使我们能够通过Hpa II和Msp I限制性内切酶的组合，独立而详细地分析活跃和不活跃的rRNA基因拷贝的甲基化模式。我们发现CpG甲基化主要存在于非活性rRNA基因拷贝的增强子和启动子区域。位于启动子区域的单个HPA II位点的甲基化与转录活性显示出特别强的相关性。
By using formaldehyde cross-linking of histones to DNA and gel retardation assays we show that formaldehyde fixation, similar to previously established psoralen photocross-linking, discriminates between nucleosome- packed (inactive) and nucleosome-free (active) fractions of ribosomal RNA genes. By both cross-linking techniques we were able to purify fragments from agarose gels, corresponding to coding, enhancer and promoter sequences of rRNA genes, which were further investigated with respect to DNA methylation. This approach allows us to analyse independently and in detail methylation patterns of active and inactive rRNA gene copies by the combination of Hpa II and Msp I restriction enzymes. We found CpG methylation mainly present in enhancer and promoter regions of inactive rRNA gene copies. The methylation of one single Hpa II site, located in the promoter region, showed particularly strong correlation with the transcriptional activity.
DOI：
10.1073/pnas.82.19.6470
发表时间：
1985-01-01
影响因子：
11.1
作者：
SOLOMON, MJ;VARSHAVSKY, A
通讯作者：
VARSHAVSKY, A
非洲爪蟾 rDNA 中 HpaII 位点的体外甲基化不影响其在卵母细胞中的转录。
影响因子：
14.9
作者：
Pennock,DG;Reeder,RH
通讯作者：
Reeder,RH