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Unwinding of a eukaryotic origin of replication visualized by cryo-EM.
Henrikus, Sarah S; Gross, Marta H; Willhoft, Oliver; Pühringer, Thomas; Lewis, Jacob S; McClure, Allison W; Greiwe, Julia F; Palm, Giacomo; Nans, Andrea; Diffley, John F X; Costa, Alessandro.
Afiliación
  • Henrikus SS; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • Gross MH; Genome Stability Unit, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia.
  • Willhoft O; Chromosome Replication Laboratory, Francis Crick Institute, London, UK.
  • Pühringer T; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • Lewis JS; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • McClure AW; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • Greiwe JF; Chromosome Replication Laboratory, Francis Crick Institute, London, UK.
  • Palm G; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
  • Nans A; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • Diffley JFX; Macromolecular Machines Laboratory, Francis Crick Institute, London, UK.
  • Costa A; Structural Biology Science Technology Platform, Francis Crick Institute, London, UK.
Nat Struct Mol Biol ; 31(8): 1265-1276, 2024 Aug.
Article en En | MEDLINE | ID: mdl-38760633
ABSTRACT
To prevent detrimental chromosome re-replication, DNA loading of a double hexamer of the minichromosome maintenance (MCM) replicative helicase is temporally separated from DNA unwinding. Upon S-phase transition in yeast, DNA unwinding is achieved in two

steps:

limited opening of the double helix and topological separation of the two DNA strands. First, Cdc45, GINS and Polε engage MCM to assemble a double CMGE with two partially separated hexamers that nucleate DNA melting. In the second step, triggered by Mcm10, two CMGEs separate completely, eject the lagging-strand template and cross paths. To understand Mcm10 during helicase activation, we used biochemical reconstitution with cryogenic electron microscopy. We found that Mcm10 splits the double CMGE by engaging the N-terminal homo-dimerization face of MCM. To eject the lagging strand, DNA unwinding is started from the N-terminal side of MCM while the hexamer channel becomes too narrow to harbor duplex DNA.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Saccharomyces cerevisiae / Origen de Réplica / Microscopía por Crioelectrón / Proteínas de Saccharomyces cerevisiae / Replicación del ADN / Proteínas de Mantenimiento de Minicromosoma Idioma: En Revista: Nat Struct Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Saccharomyces cerevisiae / Origen de Réplica / Microscopía por Crioelectrón / Proteínas de Saccharomyces cerevisiae / Replicación del ADN / Proteínas de Mantenimiento de Minicromosoma Idioma: En Revista: Nat Struct Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2024 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos