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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Autores principales:	Algirdas Toleikis, Martin R. Webb, Justin E. Molloy
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5247f0a763f74d18a7499e61c6092437
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spelling	oai:doaj.org-article:5247f0a763f74d18a7499e61c60924372021-12-02T15:08:28ZoriD structure controls RepD initiation during rolling-circle replication10.1038/s41598-017-18817-62045-2322https://doaj.org/article/5247f0a763f74d18a7499e61c60924372018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-18817-6https://doaj.org/toc/2045-2322Abstract 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.Algirdas ToleikisMartin R. WebbJustin E. MolloyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Algirdas Toleikis Martin R. Webb Justin E. Molloy oriD structure controls RepD initiation during rolling-circle replication
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.
format	article
author	Algirdas Toleikis Martin R. Webb Justin E. Molloy
author_facet	Algirdas Toleikis Martin R. Webb Justin E. Molloy
author_sort	Algirdas Toleikis
title	oriD structure controls RepD initiation during rolling-circle replication
title_short	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_sort	orid structure controls repd initiation during rolling-circle replication
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/5247f0a763f74d18a7499e61c6092437
work_keys_str_mv	AT algirdastoleikis oridstructurecontrolsrepdinitiationduringrollingcirclereplication AT martinrwebb oridstructurecontrolsrepdinitiationduringrollingcirclereplication AT justinemolloy oridstructurecontrolsrepdinitiationduringrollingcirclereplication
_version_	1718388120301338624

oriD structure controls RepD initiation during rolling-circle replication

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