Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Annu Rev Biophys. 2010;39:429-48. doi: 10.1146/annurev.biophys.093008.131327.
Replication of DNA is carried out by the replisome, a multiprotein complex responsible for the unwinding of parental DNA and the synthesis of DNA on each of the two DNA strands. The impressive speed and processivity with which the replisome duplicates DNA are a result of a set of tightly regulated interactions between the replication proteins. The transient nature of these protein interactions makes it challenging to study the dynamics of the replisome by ensemble-averaging techniques. This review describes single-molecule methods that allow the study of individual replication proteins and their functioning within the replisome. The ability to mechanically manipulate individual DNA molecules and record the dynamic behavior of the replisome while it duplicates DNA has led to an improved understanding of the molecular mechanisms underlying DNA replication.
DNA 的复制是由复制体(replicisome)完成的,它是一种负责解开亲本 DNA 并在两条 DNA 链上合成 DNA 的多蛋白复合物。复制体复制 DNA 的速度和效率令人印象深刻,这是由于复制蛋白之间一系列严格调控的相互作用的结果。这些蛋白质相互作用的瞬时性质使得通过整体平均技术研究复制体的动力学具有挑战性。这篇综述描述了单分子方法,这些方法允许研究单个复制蛋白及其在复制体中的功能。能够机械地操作单个 DNA 分子并记录复制体在复制 DNA 时的动态行为,这使得人们对 DNA 复制的分子机制有了更好的理解。