title: Post-licensing Specification of Eukaryotic Replication Origins by Facilitated Mcm2-7 Sliding along DNA.
abstract: Eukaryotic genomes are replicated from many origin sites that are licensed by the loading of the replicative DNA helicase, Mcm2-7. How eukaryotic origin positions are specified remains elusive. Here we show that, contrary to the bacterial paradigm, eukaryotic replication origins are not irrevocably defined by selection of the helicase loading site, but can shift in position after helicase loading. Using purified proteins we show that DNA translocases, including RNA polymerase, can push budding yeast Mcm2-7 double hexamers along DNA. Displaced Mcm2-7 double hexamers support DNA replication initiation distal to the loading site in vitro. Similarly, in yeast cells that are defective for transcription termination, collisions with RNA polymerase induce a redistribution of Mcm2-7 complexes along the chromosomes, resulting in a corresponding shift in DNA replication initiation sites. These results reveal a eukaryotic origin specification mechanism that departs from the classical replicon model, helping eukaryotic cells to negotiate transcription-replication conflict.
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Julien
authors.initials: J
authors.lastname: Gros
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Charanya
authors.initials: C
authors.lastname: Kumar
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Gerard
authors.initials: G
authors.lastname: Lynch
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA; Weill-Cornell Graduate School of Medical Sciences, New York, NY 10065, USA.
authors.forename: Tejas
authors.initials: T
authors.lastname: Yadav
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Iestyn
authors.initials: I
authors.lastname: Whitehouse
authors.affiliation: Molecular Biology Program, Memorial Sloan-Kettering Cancer Center (MSKCC), 1275 York Avenue, New York, NY 10065, USA. Electronic address: remusd@mskcc.org.
authors.forename: Dirk
authors.initials: D
authors.lastname: Remus
mesh_terms: D004265:DNA Helicases / Q000378:metabolism; D004261:DNA Replication; D004271:DNA, Fungal / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D064110:Minichromosome Maintenance Proteins / Q000378:metabolism*; D018741:Replication Origin; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; D012321:DNA-Directed RNA Polymerases; D004265:DNA Helicases; D064110:Minichromosome Maintenance Proteins