用于全基因组映射的DNA-蛋白质交叉链接测序
Feng Tang1,2, Jun Yuan1, Bi-Feng Yuan2
1Department of Chemistry, University of California, Riverside, Riverside, California 92521-0403, United States.
Journal of the American Chemical Society
|January 3, 2022
概括
使用一种新的DNA-蛋白质交叉链接测序 (DPC-Seq) 方法绘制了全基因组图谱. 在活跃的基因中,Tg被耗尽,但在异性染色质中被丰富,为DNA修复和基因组组织提供了洞察力.
科学领域:
- 分子生物学
- 基因组学
- DNA 修复
背景情况:
- 蒂米丁糖醇 (Tg) 是一种普遍存在的氧化DNA损伤,由蒂米丁或5-甲基-2'-脱氧提丁的氧化和脱胺引起.
- 基因组范围内的Tg分布在很大程度上是未知的.
- 了解Tg分布对于阐明它在DNA损伤和修复中的作用至关重要.
研究的目的:
- 在人类细胞中开发和应用一种全基因组图谱的新方法.
- 研究Tg的基因组分布和定位.
- 了解DNA修复酶对Tg积累的影响.
主要方法:
- 使用DNA糖酶NTHL1进行Tg的特定标记的DNA-蛋白交联测序 (DPC-Seq) 方法的开发.
- 通过DPC拉低,SDS-PAGE分离和膜转移来丰富含Tg的DNA.
- 人类细胞中Tg位点的全基因组映射,包括对NTHL1和NEIL1DNA糖酶的双重切除实验.
主要成果:
- 使用DPC-Seq在人类基因组中检测到数千个Tg位点.
- 两次切除NTHL1和NEIL1显著增加了Tg峰值,证实了它们在Tg修复中的作用.
- 发现Tg在活跃转录的区域被耗尽,并在核细胞结合部位被丰富,特别是在H3K9me2标记的异染色体中.
结论:
- DPC-Seq方法提供了一个高效的丰富和全基因组映射Tg含DNA的强大工具.
- Tg 呈现出不同的基因组定位模式,在活跃的转录区域中表现不足,在异色素中表现过多.
- 这项研究为Tg的未来功能研究提供了有价值的方法,并可用于绘制其他修饰的核酸.
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