细胞周期中与人类DNA复制起点结合的蛋白质的定位
Localization of proteins bound to a replication origin of human DNA along the cell cycle.
作者信息
Abdurashidova Gulnara, Danailov Miltcho B, Ochem Alexander, Triolo Gianluca, Djeliova Vera, Radulescu Sorina, Vindigni Alessandro, Riva Silvano, Falaschi Arturo
机构信息
International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34012 Trieste, Italy.
出版信息
EMBO J. 2003 Aug 15;22(16):4294-303. doi: 10.1093/emboj/cdg404.
Abstract
The proteins bound in vivo at the human lamin B2 DNA replication origin and their precise sites of binding were investigated along the cell cycle utilizing two novel procedures based on immunoprecipitation following UV irradiation with a pulsed laser light source. In G(1), the pre-replicative complex contains CDC6, MCM3, ORC1 and ORC2 proteins; of these, the post-replicative complex in S phase contains only ORC2; in M phase none of them are bound. The precise nucleotide of binding was identified for the two ORC and the CDC6 proteins near the start sites for leading-strand synthesis; the transition from the pre- to the post-replicative complex is accompanied by a 17 bp displacement of the ORC2 protein towards the start site.
摘要
利用基于脉冲激光光源紫外线照射后免疫沉淀的两种新方法，在细胞周期中研究了人lamin B2 DNA复制起点在体内结合的蛋白质及其精确结合位点。在G(1)期，复制前复合物包含CDC6、MCM3、ORC1和ORC2蛋白；其中，S期的复制后复合物仅包含ORC2；在M期，它们均未结合。确定了前导链合成起始位点附近两种ORC和CDC6蛋白的精确结合核苷酸；从复制前复合物到复制后复合物的转变伴随着ORC2蛋白向起始位点的17 bp位移。
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