ArticleCurrent genetics2014
The mutagenic footprint of low-fidelity Pol I ColE1 plasmid replication in E. coli reveals an extensive interplay between Pol I and Pol III.
Article in Current genetics, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 13 citations in OpenAlex.
- Ultrahigh-throughput screening-assisted in vivo directed evolution for enzyme engineering.Biotechnology for biofuels and bioproducts · 2024Article
- Mechanisms of Theta Plasmid Replication in Enterobacteria and Implications for Adaptation to Its Host.EcoSal Plus · 2020Review
- rAAV Engineering for Capsid-Protein Enzyme Insertions and Mosaicism Reveals Resilience to Mutational, Structural and Thermal Perturbations.International journal of molecular sciences · 2019Article
- Fluorescence-Based Reporters for Detection of Mutagenesis in E. coli.Methods in enzymology · 2017Article
- Development of potent in vivo mutagenesis plasmids with broad mutational spectra.Nature communications · 2015Article
- Mechanisms of Theta Plasmid Replication.Microbiology spectrum · 2015Review
- Random mutagenesis by error-prone pol plasmid replication in Escherichia coli.Methods in molecular biology (Clifton, N.J.) · 2014Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
Abstract
ColE1 plasmid replication is unidirectional and requires two DNA polymerases: DNA polymerase I (Pol I) and DNA polymerase III (Pol III). Pol I initiates leading-strand synthesis by extending an RNA primer, allowing the Pol III holoenzyme to assemble and finish replication of both strands. The goal of the present work is to study the interplay between Pol I and Pol III during ColE1 plasmid replication, to gain new insights into Pol I function in vivo. Our approach consists of using mutations generated by a low-fidelity mutant of Pol I (LF-Pol I) during replication of a ColE1 plasmid as a footprint for Pol I replication. This approach allowed mapping areas of Pol I replication on the plasmid with high resolution. In addition, we were able to approximate the strandedness of Pol I mutations throughout the plasmid, allowing us to estimate the spectrum of the LF-Pol I in vivo. Our study produced the following three mechanistic insights: (1) we identified the likely location of the polymerase switch at ~200 bp downstream of replication initiation; (2) we found evidence suggesting that Pol I can replicate both strands, supporting earlier studies indicating a functional redundancy between Pol I and Pol III (3) we found evidence pointing to a specific role of Pol I during termination of lagging-strand replication. In addition, we illustrate how our strand-specific footprinting approach can be used to dissect factors modulating Pol I fidelity in vivo.
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Registered trials
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