oriD structure controls RepD initiation during rolling-circle replication View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2018-12

AUTHORS

Algirdas Toleikis, Martin R. Webb, Justin E. Molloy

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. More... »

PAGES

1206

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-017-18817-6

DOI

http://dx.doi.org/10.1038/s41598-017-18817-6

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1100406225

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/29352198


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