title: Cryo-EM structure of the replisome reveals multiple interactions coordinating DNA synthesis.
abstract: We present a structure of the ∼650-kDa functional replisome of bacteriophage T7 assembled on DNA resembling a replication fork. A structure of the complex consisting of six domains of DNA helicase, five domains of RNA primase, two DNA polymerases, and two thioredoxin (processivity factor) molecules was determined by single-particle cryo-electron microscopy. The two molecules of DNA polymerase adopt a different spatial arrangement at the replication fork, reflecting their roles in leading- and lagging-strand synthesis. The structure, in combination with biochemical data, reveals molecular mechanisms for coordination of leading- and lagging-strand synthesis. Because mechanisms of DNA replication are highly conserved, the observations are relevant to other replication systems.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; arek@hms.harvard.edu ccr@hms.harvard.edu.
authors.forename: Arkadiusz W
authors.initials: AW
authors.lastname: Kulczyk
authors.affiliation: Department of Molecular Biology and Genetics, The Danish Research Institute of Translational Neuroscience, Aarhus University, 8000 Aarhus, Denmark.
authors.forename: Arne
authors.initials: A
authors.lastname: Moeller
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
authors.forename: Peter
authors.initials: P
authors.lastname: Meyer
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
authors.forename: Piotr
authors.identifier: 0000-0002-6522-0835
authors.initials: P
authors.lastname: Sliz
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115; arek@hms.harvard.edu ccr@hms.harvard.edu.
authors.forename: Charles C
authors.initials: CC
authors.lastname: Richardson
mesh_terms: D017123:Bacteriophage T7 / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D020285:Cryoelectron Microscopy; D004247:DNA / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D004261:DNA Replication / Q000235:genetics*; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D000072417:Protein Domains; D013879:Thioredoxins / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure
chemical_list: D009097:Multienzyme Complexes; D013879:Thioredoxins; D004247:DNA
keywords: DNA polymerase; DNA replication; coordination of leading- and lagging-strands synthesis; cryo-EM structure; replisome