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oriD structure controls RepD initiation during rolling-circle replication
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Toleikis, Algirdas, Webb, Martin R. and Molloy, Justin E. (2018) oriD structure controls RepD initiation during rolling-circle replication. Scientific Reports, 8 (1). p. 1206. 1206. doi:10.1038/s41598-017-18817-6
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Official URL: https://doi.org/10.1038/s41598-017-18817-6
Abstract
Bacterial antibiotic resistance is often carried by circular DNA plasmids that are copied separately from the genomic DNA and can be passed to other bacteria, spreading the resistance. The chloramphenicol-resistance plasmid pC221 from Staphylococcus aureus is duplicated by a process called asymmetric rolling circle replication. It is not fully understood how the replication process is regulated but its initiation requires a plasmid-encoded protein called RepD that nicks one strand of the parent plasmid at the double-stranded origin of replication (oriD). Using magnetic tweezers to control the DNA linking number we found RepD nicking occurred only when DNA was negatively supercoiled and that binding of a non-nicking mutant (RepDY188F) stabilized secondary structure formation at oriD. Quenched-flow experiments showed the inverted complementary repeat sequence, ICRII, within oriD was most important for rapid nicking of intact plasmids. Our results show that cruciform formation at oriD is an important control for initiation of plasmid replication.
| Item Type: | Journal Article | ||||||
|---|---|---|---|---|---|---|---|
| Divisions: | Faculty of Medicine > Warwick Medical School | ||||||
| SWORD Depositor: | Library Publications Router | ||||||
| Journal or Publication Title: | Scientific Reports | ||||||
| Publisher: | Nature Publishing Group | ||||||
| ISSN: | 2045-2322 | ||||||
| Official Date: | 19 January 2018 | ||||||
| Dates: |
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| Volume: | 8 | ||||||
| Number: | 1 | ||||||
| Page Range: | p. 1206 | ||||||
| Article Number: | 1206 | ||||||
| DOI: | 10.1038/s41598-017-18817-6 | ||||||
| Status: | Peer Reviewed | ||||||
| Publication Status: | Published | ||||||
| Publisher Statement: | ** From PubMed via Jisc Publications Router. ** History: received 07-09-2017; accepted 18-12-2017. | ||||||
| Access rights to Published version: | Open Access |
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