Cryo-EM of dynamic protein complexes in eukaryotic DNA replication.

ABSTRACT: DNA replication in Eukaryotes is a highly dynamic process that involves several dozens of proteins. Some of these proteins form stable complexes that are amenable to high-resolution structure determination by cryo-EM, thanks to the recent advent of the direct electron detector and powerful image analysis algorithm. But many of these proteins associate only transiently and flexibly, precluding traditional biochemical purification. We found that direct mixing of the component proteins followed by 2D and 3D image sorting can capture some very weakly interacting complexes. Even at 2D average level and at low resolution, EM images of these flexible complexes can provide important biological insights. It is often necessary to positively identify the feature-of-interest in a low resolution EM str

SUBMITTER: Sun J 

PROVIDER: S-EPMC5192969 | biostudies-literature | 2017 Jan

REPOSITORIES: biostudies-literature