Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli Journal Article
| Authors: | Gupta, M. K.; Guy, C. P.; Yeeles, J. T. P.; Atkinson, J.; Bell, H.; Lloyd, R. G.; Marians, K. J.; McGlynn, P. |
| Article Title: | Protein-DNA complexes are the primary sources of replication fork pausing in Escherichia coli |
| Abstract: | Replication fork pausing drives genome instability, because any loss of paused replisome activity creates a requirement for reloading of the replication machinery, a potentially mutagenic process. Despite this importance, the relative contributions to fork pausing of different replicative barriers remain unknown. We show here that Deinococcus radiodurans RecD2 helicase inactivates Escherichia coli replisomes that are paused but still functional in vitro, preventing continued fork movement upon barrier removal or bypass, but does not inactivate elongating forks. Using RecD2 to probe replisome pausing in vivo, we demonstrate that most pausing events do not lead to replisome inactivation, that transcription complexes are the primary sources of this pausing, and that an accessory replicative helicase is critical for minimizing the frequency and/or duration of replisome pauses. These findings reveal the hidden potential for replisome inactivation, and hence genome instability, inside cells. They also demonstrate that efficient chromosome duplication requires mechanisms that aid resumption of replication by paused replisomes, especially those halted by protein- DNA barriers such as transcription complexes. |
| Keywords: | unclassified drug; dna-binding proteins; nonhuman; dna replication; cell death; dna repair; protein binding; in vivo study; transcription, genetic; in vitro study; genomic instability; escherichia coli; helicase; multienzyme complexes; dna, bacterial; dna helicases; escherichia coli proteins; genome stability; rna polymerase; recombination; dna-directed dna polymerase; replisome; deinococcus radiodurans; dna protein complex; chromosome duplication; rep; recd2 helicase; nucleoproteins |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 110 |
| Issue: | 18 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 2013-04-30 |
| Start Page: | 7252 |
| End Page: | 7257 |
| Language: | English |
| DOI: | 10.1073/pnas.1303890110 |
| PROVIDER: | scopus |
| PMCID: | PMC3645559 |
| PUBMED: | 23589869 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 3 June 2013" - "CODEN: PNASA" - "Source: Scopus" |
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