Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
Nucleic Acids Res. 2012 Jul;40(12):5487-96. doi: 10.1093/nar/gks217. Epub 2012 Mar 8.
Analyses of the DNA replication-associated proteins of hyperthermophilic archaea have yielded considerable insight into the structure and biochemical function of these evolutionarily conserved factors. However, little is known about the regulation and progression of DNA replication in the context of archaeal cells. In the current work, we describe the generation of strains of Sulfolobus solfataricus and Sulfolobus acidocaldarius that allow the incorporation of nucleoside analogues during DNA replication. We employ this technology, in conjunction with immunolocalization analyses of replisomes, to investigate the sub-cellular localization of nascent DNA and replisomes. Our data reveal a peripheral localization of replisomes in the cell. Furthermore, while the two replication forks emerging from any one of the three replication origins in the Sulfolobus chromosome remain in close proximity, the three origin loci are separated.
对嗜热古菌的 DNA 复制相关蛋白的分析为这些进化上保守的因子的结构和生化功能提供了重要的认识。然而，关于古菌细胞中 DNA 复制的调控和进程知之甚少。在目前的工作中，我们描述了生成能够在 DNA 复制过程中掺入核苷类似物的 Sulfolobus solfataricus 和 Sulfolobus acidocaldarius 菌株的方法。我们结合复制体的免疫定位分析，利用这项技术来研究新生 DNA 和复制体的亚细胞定位。我们的数据揭示了复制体在细胞中的外周定位。此外，尽管 Sulfolobus 染色体上的三个复制起点中的任何一个产生的两个复制叉仍然保持紧密接近，但三个原点位点是分开的。
