Localization of proteins bound to a replication origin of human DNA along the cell cycle
Localization of proteins bound to a replication origin of human DNA along the cell cycle
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DOI：
10.1093/emboj/cdg404
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发表时间：
2003-08-15
期刊：
影响因子：
11.4
通讯作者：
Falaschi, A
中科院分区：
文献类型：
--
作者：
Abdurashidova, G;Danailov, MB;Falaschi, A
利用两种基于免疫沉淀的新方法，在用脉冲激光光源进行UV照射后，研究了在体内结合在人核纤层蛋白B2 DNA复制起点和其精确结合位点的蛋白质沿细胞周期的沿着。在G（1）中，复制前复合物含有CDC 6、MCM 3、ORC 1和ORC 2蛋白;其中，S期的复制后复合物仅含有ORC 2;在M期，它们都不结合。精确的核苷酸的结合被确定为两个ORC和CDC 6蛋白附近的起始位点的前导链合成;从复制前到复制后复合物的过渡伴随着一个17 bp的位移的ORC 2蛋白朝向起始位点。
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.