Nucleotide binding halts diffusion of the eukaryotic replicative helicase during activation.

ABSTRACT: The eukaryotic replicative helicase CMG centrally orchestrates the replisome and leads the way at the front of replication forks. Understanding the motion of CMG on the DNA is therefore key to our understanding of DNA replication. In vivo, CMG is assembled and activated through a cell-cycle-regulated mechanism involving 36 polypeptides that has been reconstituted from purified proteins in ensemble biochemical studies. Conversely, single-molecule studies of CMG motion have thus far relied on pre-formed CMG assembled through an unknown mechanism upon overexpression of individual constituents. Here, we report the activation of CMG fully reconstituted from purified yeast proteins and the quantification of its motion at the single-molecule level. We observe that CMG can move on DNA in two ways:

SUBMITTER: Ramirez Montero D 

PROVIDER: S-EPMC10104875 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature