A Primase-Induced Conformational Switch Controls the Stability of the Bacterial Replisome

Authors

Enrico Monachino, University of Wollongong
Slobodan Jergic, University of WollongongFollow
Jacob S. Lewis, University of WollongongFollow
Zhi-Qiang Xu, University of WollongongFollow
Allen Lo, University of Wollongong
Valerie O'Shea
James Berger
Nicholas E. Dixon, University of WollongongFollow
Antoine M. van Oijen, University of WollongongFollow

RIS ID

143874

Publication Details

Monachino, E., Jergic, S., Lewis, J., Xu, Z., Lo, A., O'Shea, V., Berger, J., Dixon, N. & van Oijen, A. (2020). A Primase-Induced Conformational Switch Controls the Stability of the Bacterial Replisome. Molecular Cell,

Abstract

© 2020 Elsevier Inc. Recent studies of bacterial DNA replication have led to a picture of the replisome as an entity that freely exchanges DNA polymerases and displays intermittent coupling between the helicase and polymerase(s). Challenging the textbook model of the polymerase holoenzyme acting as a stable complex coordinating the replisome, these observations suggest a role of the helicase as the central organizing hub. We show here that the molecular origin of this newly found plasticity lies in the 500-fold increase in strength of the interaction between the polymerase holoenzyme and the replicative helicase upon association of the primase with the replisome. By combining in vitro ensemble-averaged and single-molecule assays, we demonstrate that this conformational switch operates during replication and promotes recruitment of multiple holoenzymes at the fork. Our observations provide a molecular mechanism for polymerase exchange and offer a revised model for the replication reaction that emphasizes its stochasticity.