oriD structure controls RepD initiation during rolling-circle replication
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...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2018-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-18817-6 |
| _version_ | 1818996594477891584 |
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| author | Algirdas Toleikis Martin R. Webb Justin E. Molloy |
| author_facet | Algirdas Toleikis Martin R. Webb Justin E. Molloy |
| author_sort | Algirdas Toleikis |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-20T21:32:13Z |
| format | Article |
| id | doaj.art-5247f0a763f74d18a7499e61c6092437 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T21:32:13Z |
| publishDate | 2018-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-5247f0a763f74d18a7499e61c60924372022-12-21T19:26:01ZengNature PortfolioScientific Reports2045-23222018-01-018111210.1038/s41598-017-18817-6oriD structure controls RepD initiation during rolling-circle replicationAlgirdas Toleikis0Martin R. Webb1Justin E. Molloy2The Francis Crick InstituteThe Francis Crick InstituteThe Francis Crick InstituteAbstract 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.https://doi.org/10.1038/s41598-017-18817-6 |
| spellingShingle | Algirdas Toleikis Martin R. Webb Justin E. Molloy oriD structure controls RepD initiation during rolling-circle replication Scientific Reports |
| title | oriD structure controls RepD initiation during rolling-circle replication |
| title_full | oriD structure controls RepD initiation during rolling-circle replication |
| title_fullStr | oriD structure controls RepD initiation during rolling-circle replication |
| title_full_unstemmed | oriD structure controls RepD initiation during rolling-circle replication |
| title_short | oriD structure controls RepD initiation during rolling-circle replication |
| title_sort | orid structure controls repd initiation during rolling circle replication |
| url | https://doi.org/10.1038/s41598-017-18817-6 |
| work_keys_str_mv | AT algirdastoleikis oridstructurecontrolsrepdinitiationduringrollingcirclereplication AT martinrwebb oridstructurecontrolsrepdinitiationduringrollingcirclereplication AT justinemolloy oridstructurecontrolsrepdinitiationduringrollingcirclereplication |