Structures of pMV158 replication initiator RepB with and without DNA reveal a flexible dual-function protein

DNA replication is essential to all living organisms as it ensures the fidelity of genetic material for the next generation of dividing cells. One of the simplest replication initiation mechanisms is the rolling circle replication. In the streptococcal plasmid pMV158, which confers antibiotic resist...

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Detalles Bibliográficos
Autores: Machón Sobrado, Cristina, Ruiz Masó, José Angel, Amodio, Juliana, Boer. D. Roeland, Bordanaba Ruiseco, Lorena, Bury, Katarzyna, Konieczny, Igor, Solar, Gloria del, Coll Capella, Miquel, 1955-
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/200288
Acceso en línea:https://hdl.handle.net/2445/200288
Access Level:acceso abierto
Palabra clave:Duplicació de l'ADN
Plasmidis
DNA replication
Plasmids
Descripción
Sumario:DNA replication is essential to all living organisms as it ensures the fidelity of genetic material for the next generation of dividing cells. One of the simplest replication initiation mechanisms is the rolling circle replication. In the streptococcal plasmid pMV158, which confers antibiotic resistance to tetracycline, replication initiation is catalysed by RepB protein. The RepB N-terminal domain or origin binding domain binds to the recognition sequence (bind locus) of the double-strand origin of replication and cleaves one DNA strand at a specific site within the nic locus. Using biochemical and crystallographic analyses, here we show how the origin binding domain recognises and binds to the bind locus using structural elements removed from the active site, namely the recognition α helix, and a β-strand that organises upon binding. A new hexameric structure of full-length RepB that highlights the great flexibility of this protein is presented, which could account for its ability to perform different tasks, namely bind to two distinct loci and cleave one strand of DNA at the plasmid origin.© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.