title: CRL2Lrr1promotes unloading of the vertebrate replisome from chromatin during replication termination.
abstract: A key event during eukaryotic replication termination is the removal of the CMG helicase from chromatin. CMG unloading involves ubiquitylation of its Mcm7 subunit and the action of the p97 ATPase. Using a proteomic screen inXenopusegg extracts, we identified factors that are enriched on chromatin when CMG unloading is blocked. This approach identified the E3 ubiquitin ligase CRL2Lrr1, a specific p97 complex, other potential regulators of termination, and many replisome components. We show that Mcm7 ubiquitylation and CRL2Lrr1binding to chromatin are temporally linked and occur only during replication termination. In the absence of CRL2Lrr1, Mcm7 is not ubiquitylated, CMG unloading is inhibited, and a large subcomplex of the vertebrate replisome that includes DNA Pol ε is retained on DNA. Our data identify CRL2Lrr1as a master regulator of replisome disassembly during vertebrate DNA replication termination.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: James M
authors.initials: JM
authors.lastname: Dewar
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Low
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
authors.affiliation: Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Räschle
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.|Howard Hughes Medical Institute, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Johannes C
authors.initials: JC
authors.lastname: Walter
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004265:DNA Helicases / Q000378:metabolism*; D004257:DNA Polymerase II / Q000378:metabolism; D004261:DNA Replication; D064168:Minichromosome Maintenance Complex Component 7 / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics* / Q000378:metabolism
chemical_list: D002843:Chromatin; D009687:Nuclear Proteins; D029867:Xenopus Proteins; D004247:DNA; D044767:Ubiquitin-Protein Ligases; D004257:DNA Polymerase II; D000251:Adenosine Triphosphatases; C404942:p97 ATPase; D004265:DNA Helicases; C509836:Mcm7 protein, Xenopus; D064168:Minichromosome Maintenance Complex Component 7
keywords: CMG; DNA replication; p97; replication termination; ubiquitin