The human GINS complex binds to and specifically stimulates human DNA polymerase α-primase
The human GINS complex binds to and specifically stimulates human DNA polymerase α-primase
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DOI：
10.1038/sj.embor.7400870
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发表时间：
2007-01-01
期刊：
影响因子：
7.7
通讯作者：
Pisani, Francesca M.
中科院分区：
文献类型：
--
作者：
De Falco, Mariarosaria;Ferrari, Elena;Pisani, Francesca M.
真核GINS复合物在基因组复制的起始和延伸阶段具有重要作用。它由四个旁系同源的亚基组成-Sld 5，Psf 1，Psf 2和Psf 3-它们在真核生物中普遍存在且进化上保守。在这里，我们报告的人GINS复合物（hGINS）的生化特性。四个hGINS亚基在大肠杆菌中以高度可溶的形式共表达，并作为复合物纯化。通过使用表面等离子体共振测量或通过用抗hGINS抗体进行的免疫沉淀实验，hGINS显示出与异二聚体人DNA引发酶直接相互作用。DNA聚合酶α-引发酶的合成活性被hGINS在各种引发的DNA模板上特异性地刺激。这些研究结果的意义进行了讨论，鉴于在人类复制叉的分子动力学。
The eukaryotic GINS complex has an essential role in the initiation and elongation phases of genome duplication. It is composed of four paralogous subunits-Sld5, Psf1, Psf2 and Psf3-which are ubiquitous and evolutionarily conserved in eukaryotic organisms. Here, we report the biochemical characterization of the human GINS complex (hGINS). The four hGINS subunits were coexpressed in Escherichia coli in a highly soluble form and purified as a complex. hGINS was shown to interact directly with the heterodimeric human DNA primase, by using either surface plasmon resonance measurements or by immunoprecipitation experiments carried out with anti-hGINS antibodies. The DNA polymerase alpha-primase synthetic activity was specifically stimulated by hGINS on various primed DNA templates. The significance of these findings is discussed in view of the molecular dynamics at the human replication fork.