TY - JOUR TI - Transcription leads to pervasive replisome instability in bacteria AU - Mangiameli, Sarah M AU - Merrikh, Christopher N AU - Wiggins, Paul A AU - Merrikh, Houra A2 - van Oijen, Antoine M VL - 6 PY - 2017 DA - 2017/01/16 SP - e19848 C1 - eLife 2017;6:e19848 DO - 10.7554/eLife.19848 UR - https://doi.org/10.7554/eLife.19848 AB - The canonical model of DNA replication describes a highly-processive and largely continuous process by which the genome is duplicated. This continuous model is based upon in vitro reconstitution and in vivo ensemble experiments. Here, we characterize the replisome-complex stoichiometry and dynamics with single-molecule resolution in bacterial cells. Strikingly, the stoichiometries of the replicative helicase, DNA polymerase, and clamp loader complexes are consistent with the presence of only one active replisome in a significant fraction of cells (>40%). Furthermore, many of the observed complexes have short lifetimes (<8 min), suggesting that replisome disassembly is quite prevalent, possibly occurring several times per cell cycle. The instability of the replisome complex is conflict-induced: transcription inhibition stabilizes these complexes, restoring the second replisome in many of the cells. Our results suggest that, in contrast to the canonical model, DNA replication is a largely discontinuous process in vivo due to pervasive replication-transcription conflicts. KW - replisome KW - DNA replication KW - replication-transcription conflicts KW - replication rates JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -