Localization of Bacterial DNA Polymerase: Evidence for a Factory Model of Replication

TLDR

DNA replication has been described by two models: a moving polymerase model and a stationary polymerase factory model in which DNA is pulled through. The study aims to distinguish between the moving and stationary polymerase models. The authors visualized Bacillus subtilis DNA polymerase by fusing its catalytic subunit PolC to GFP and observing its localization in living cells. PolC‑GFP localized to discrete midcell positions, indicating the polymerase is anchored and supporting the stationary factory model.

Abstract

Two general models have been proposed for DNA replication. In one model, DNA polymerase moves along the DNA (like a train on a track); in the other model, the polymerase is stationary (like a factory), and DNA is pulled through. To distinguish between these models, we visualized DNA polymerase of the bacterium Bacillus subtilis in living cells by the creation of a fusion protein containing the catalytic subunit (PolC) and green fluorescent protein (GFP). PolC-GFP was localized at discrete intracellular positions, predominantly at or near midcell, rather than being distributed randomly. These results suggest that the polymerase is anchored in place and thus support the model in which the DNA template moves through the polymerase.