Identification of novel initiation sites for human DNA replication around ARSH1, a previously characterized yeast replicator
Identification of novel initiation sites for human DNA replication around ARSH1, a previously characterized yeast replicator
复制标题
DOI：
10.1016/j.bbrc.2003.12.040
复制
发表时间：
2004-01-23
影响因子：
3.1
通讯作者：
Valenzuela, MS
中科院分区：
文献类型：
--
作者：
Hu, L;Xu, X;Valenzuela, MS
哺乳动物染色体的复制依赖于沿染色体分布的大量DNA复制起始点的激活。我们将注意力集中在人类DNA区域，名为ARSH1，定位于2号染色体上，之前已被证明在酵母酵母中起着附带起源的作用。在目前的研究中，我们使用了新生链DNA丰度测定来绘制体内人类染色体中包含ARSH1的5kb区域内DNA复制的起始位点。该分析应用于从正常皮肤成纤维细胞中分离的1-1.4 kb新生DNA链片段，发现在ARSH1区域周围存在两个主要起始位点。从HeLa细胞中获得等效的DNA片段，除了这些位点外，还观察到广泛的起始谱，其中包括ARSH1区域。然而，该DNA区域不足以支持转染到HeLa细胞中的含有arsh1的质粒的体外复制。(C) 2003 Elsevier Inc.版权所有。
Replication of mammalian chromosomes depends on the activation of a large number of origins of DNA replication distributed along the chromosomes. We have focused our attention on a human DNA region, named ARSH1, localized to chromosome 2, that had been previously shown to act as an episomal origin in the yeast Saccharomyces cerevisiae. In the present study we have used a nascent strand DNA abundance assay to map initiation sites for DNA replication in in vivo human chromosomes around a 5 kb region encompassing ARSH1. This analysis applied to a 1-1.4 kb nascent DNA strand fraction isolated from normal skin fibroblasts revealed the presence of two major initiations sites surrounding the ARSH1 region. With an equivalent DNA fraction obtained from HeLa cells, in addition to these sites, a broad initiation profile was observed which included the ARSH1 region. This DNA region however was not sufficient to support episomal replication of an ARSH1-containing plasmid transfected into HeLa cells. (C) 2003 Elsevier Inc. All rights reserved.