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Title: The consequences of replicating in the wrong orientation: Bacterial chromosome duplication without an active replication origin
Authors: Dimude, JU
Stockum, A
Midgley-Smith, SL
Upton, AL
Foster, HA
Khan, A
Saunders, NJ
Retkute, R
Rudolph, CJ
Keywords: DNA replication;RecG;RNase HI;Replication fork fusions;Replication-transcription conflicts;Origin-independent replication
Issue Date: 2015
Publisher: The American Society for Microbiology
Citation: mBio, 6(6): e01294-15, (2015)
Abstract: Chromosome replication is regulated in all organisms at the assembly stage of the replication machinery at specific origins. In Escherichia coli the DnaA initiator protein regulates the assembly of replication forks at oriC. This regulation can be undermined by defects in nucleic acid meta¬bolism. In cells lacking RNase HI replication initiates indepen¬dently of DnaA and oriC, presumably at persisting R-loops. A similar mechanism was assumed for origin-independent synthesis in cells lacking RecG. However, recently we suggested that this synthesis initiates at intermediates resulting from replication fork fusions. Here we present data suggesting that in cells lacking RecG or RNase HI origin-independent synthesis arises by different mechanisms, indicative of these two proteins having different roles in vivo. Our data support the idea that RNase HI processes R-loops, while RecG is required to process replication fork fusion intermediates. However, regardless of how origin-independent synthesis is initiated, a fraction of forks will proceed in an orientation opposite to normal. We show that the resulting head-on encounters with transcription threaten cell viability, especially if taking place in highly-transcribed areas. Thus, despite their different functions, RecG and RNase HI are both important factors for maintaining replication control and orientation. Their absence causes severe replication problems, highlighting the advantages of the normal chromosome arrangement, which exploits a single origin to control the number of forks and their orientation relative to transcription, and a defined termination area to contain fork fusions. Any changes to this arrangement endanger cell cycle control, chromosome dynamics and, ultimately, cell viability.
ISSN: 2161-2129
Appears in Collections:Dept of Life Sciences Research Papers

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