Structural mechanism for replication origin binding and remodeling by a metazoan origin recognition complex and its co-loader Cdc6.
AAA Domain
Adenosine Triphosphate
/ metabolism
Animals
Cell Cycle Proteins
/ chemistry
Cryoelectron Microscopy
DNA
/ chemistry
Drosophila Proteins
/ chemistry
Drosophila melanogaster
/ genetics
Hydrolysis
Minichromosome Maintenance Proteins
/ chemistry
Models, Molecular
Origin Recognition Complex
/ chemistry
Protein Binding
Recombinant Proteins
/ chemistry
Replication Origin
/ genetics
Saccharomyces cerevisiae
/ genetics
Saccharomyces cerevisiae Proteins
/ chemistry
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
26 08 2020
26 08 2020
Historique:
received:
16
06
2020
accepted:
03
08
2020
entrez:
28
8
2020
pubmed:
28
8
2020
medline:
15
9
2020
Statut:
epublish
Résumé
Eukaryotic DNA replication initiation relies on the origin recognition complex (ORC), a DNA-binding ATPase that loads the Mcm2-7 replicative helicase onto replication origins. Here, we report cryo-electron microscopy (cryo-EM) structures of DNA-bound Drosophila ORC with and without the co-loader Cdc6. These structures reveal that Orc1 and Orc4 constitute the primary DNA binding site in the ORC ring and cooperate with the winged-helix domains to stabilize DNA bending. A loop region near the catalytic Walker B motif of Orc1 directly contacts DNA, allosterically coupling DNA binding to ORC's ATPase site. Correlating structural and biochemical data show that DNA sequence modulates DNA binding and remodeling by ORC, and that DNA bending promotes Mcm2-7 loading in vitro. Together, these findings explain the distinct DNA sequence-dependencies of metazoan and S. cerevisiae initiators in origin recognition and support a model in which DNA geometry and bendability contribute to Mcm2-7 loading site selection in metazoans.
Identifiants
pubmed: 32848132
doi: 10.1038/s41467-020-18067-7
pii: 10.1038/s41467-020-18067-7
pmc: PMC7450096
doi:
Substances chimiques
Cdc6 protein, Drosophila
0
Cell Cycle Proteins
0
Drosophila Proteins
0
Origin Recognition Complex
0
Recombinant Proteins
0
Saccharomyces cerevisiae Proteins
0
Adenosine Triphosphate
8L70Q75FXE
DNA
9007-49-2
Minichromosome Maintenance Proteins
EC 3.6.4.12
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
4263Références
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