School of Biology, University of Nottingham, Queen's Medical Centre, Nottingham, NG7 2UH, UK.
DeepSeq, University of Nottingham, Queen's Medical Centre, Nottingham, NG7 2UH, UK.
Nature. 2013 Nov 28;503(7477):544-547. doi: 10.1038/nature12650. Epub 2013 Nov 3.
DNA replication initiates at defined sites called origins, which serve as binding sites for initiator proteins that recruit the replicative machinery. Origins differ in number and structure across the three domains of life and their properties determine the dynamics of chromosome replication. Bacteria and some archaea replicate from single origins, whereas most archaea and all eukaryotes replicate using multiple origins. Initiation mechanisms that rely on homologous recombination operate in some viruses. Here we show that such mechanisms also operate in archaea. We use deep sequencing to study replication in Haloferax volcanii and identify four chromosomal origins of differing activity. Deletion of individual origins results in perturbed replication dynamics and reduced growth. However, a strain lacking all origins has no apparent defects and grows significantly faster than wild type. Origin-less cells initiate replication at dispersed sites rather than at discrete origins and have an absolute requirement for the recombinase RadA, unlike strains lacking individual origins. Our results demonstrate that homologous recombination alone can efficiently initiate the replication of an entire cellular genome. This raises the question of what purpose replication origins serve and why they have evolved.
DNA 复制从称为起点的特定位置起始,这些位置作为起始蛋白的结合位点,招募复制机制。起点在生命的三个领域中的数量和结构都不同,它们的特性决定了染色体复制的动态。细菌和一些古菌从单个起点复制,而大多数古菌和所有真核生物都使用多个起点复制。依赖同源重组的起始机制在一些病毒中起作用。在这里,我们表明这种机制也在古菌中起作用。我们使用深度测序来研究 Haloferax volcanii 中的复制,并鉴定出四个具有不同活性的染色体起点。单个起点的缺失会导致复制动力学受到干扰和生长减少。然而,缺乏所有起点的菌株没有明显的缺陷,并且比野生型生长得快得多。没有起点的细胞在分散的位点而不是在离散的起点开始复制,并且与缺乏单个起点的菌株不同,它们绝对需要重组酶 RadA。我们的结果表明,同源重组可以有效地启动整个细胞基因组的复制。这就提出了一个问题,即复制起点的作用是什么,以及它们为什么会进化。