High-throughput mapping of origins of replication in human cells
High-throughput mapping of origins of replication in human cells
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DOI：
10.1038/sj.embor.7401026
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发表时间：
2007-08-01
期刊：
影响因子：
7.7
通讯作者：
Le Beau, Michelle M.
中科院分区：
文献类型：
--
作者：
Lucas, Isabelle;Palakodeti, Aparna;Le Beau, Michelle M.
在高等真核生物中，绘制复制起点的图谱一直是一个挑战。我们开发了一种快速的、全基因组的方法来绘制异步人类细胞的复制起始图，方法是将新生的链丰度分析与高平铺的微阵列平台相结合，并通过两种独立的分析验证了这项技术。我们应用这种方法分析了三个50kb区域的新生DNA的浓缩情况，这三个区域包含MYC、层蛋白B2(LMNB2)和血红蛋白β(HBB)基因的已知复制起点，一个200kb的区域包含罕见的脆性位点FRAXA，以及22号染色体上的一个1075kb的区域；我们检测到了大多数已知起点和28个新的起点。令人惊讶的是，这28个新来源的大小很小，主要位于基因内。我们的研究还表明，起始复制时间和染色质乙酰化之间存在很强的相关性。
Mapping origins of replication has been challenging in higher eukaryotes. We have developed a rapid, genome-wide method to map origins of replication in asynchronous human cells by combining the nascent strand abundance assay with a highly tiled microarray platform, and we validated the technique by two independent assays. We applied this method to analyse the enrichment of nascent DNA in three 50-kb regions containing known origins of replication in the MYC, lamin B2 ( LMNB2) and haemoglobin beta (HBB) genes, a 200-kb region containing the rare fragile site, FRAXA, and a 1,075-kb region on chromosome 22; we detected most of the known origins and also 28 new origins. Surprisingly, the 28 new origins were small in size and located predominantly within genes. Our study also showed a strong correlation between origin replication timing and chromatin acetylation.