Work

DciA, the Bacterial Replicative Helicase Loader, Promotes LLPS in the Presence of ssDNA

Year: 2024

Type: article

Abstract: • This study shows that DciA, the bacterial replicative helicase loader, promotes condensates in the presence of DNA. • This opens the way to the possibility of non-membrane compartments in bacteria. • The goal could be to concentrate the players involved in replication and thus facilitate it. The loading of the bacterial replicative helicase DnaB is an essential step for genome replication and depends on the assistance of accessory proteins. Several of these proteins have been identified across the bacterial phyla. DciA is the most common loading protein in bacteria, yet the one whose mechanism is the least understood. We have previously shown that DciA from Vibrio cholerae is composed of a globular domain followed by an unfolded extension and demonstrated its strong affinity for DNA. Here, we characterize the condensates formed by Vc DciA upon interaction with a short single-stranded DNA substrate. We demonstrate the fluidity of these condensates using light microscopy and address their network organization through electron microscopy, thereby bridging events to conclude on a liquid–liquid phase separation behavior. Additionally, we observe the recruitment of DnaB in the droplets, concomitant with the release of DciA. We show that the well-known helicase loader DnaC from Escherichia coli is also competent to form these phase-separated condensates in the presence of ssDNA. Our phenomenological data are still preliminary as regards the existence of these condensates in vivo , but open the way for exploring the potential involvement of DciA in the formation of non-membrane compartments within the bacterium to facilitate the assembly of replication players on chromosomal DNA.

Source: Journal of Molecular Biology

DOI: 10.1016/j.jmb.2024.168873