title: Role for RNA:DNA hybrids in origin-independent replication priming in a eukaryotic system.
abstract: DNA replication initiates at defined replication origins along eukaryotic chromosomes, ensuring complete genome duplication within a single S-phase. A key feature of replication origins is their ability to control the onset of DNA synthesis mediated by DNA polymerase-α and its intrinsic RNA primase activity. Here, we describe a novel origin-independent replication process that is mediated by transcription. RNA polymerase I transcription constraints lead to persistent RNA:DNA hybrids (R-loops) that prime replication in the ribosomal DNA locus. Our results suggest that eukaryotic genomes have developed tools to prevent R-loop-mediated replication events that potentially contribute to copy number variation, particularly relevant to carcinogenesis.
authors.affiliation: Department of Molecular Biology, Centro Andaluz de Biología Molecular y Medicina Regenerativa-Universidad de Sevilla, 41092 Seville, Spain.
authors.forename: Ruth
authors.initials: R
authors.lastname: Stuckey
authors.affiliation: Department of Molecular Biology, Centro Andaluz de Biología Molecular y Medicina Regenerativa-Universidad de Sevilla, 41092 Seville, Spain.
authors.forename: Néstor
authors.initials: N
authors.lastname: García-Rodríguez
authors.affiliation: Department of Molecular Biology, Centro Andaluz de Biología Molecular y Medicina Regenerativa-Universidad de Sevilla, 41092 Seville, Spain.
authors.forename: Andrés
authors.initials: A
authors.lastname: Aguilera
authors.affiliation: Department of Molecular Biology, Centro Andaluz de Biología Molecular y Medicina Regenerativa-Universidad de Sevilla, 41092 Seville, Spain ralf.wellinger@cabimer.es.
authors.forename: Ralf Erik
authors.initials: RE
authors.lastname: Wellinger
mesh_terms: D001426:Bacterial Proteins; D002166:Camptothecin / Q000737:chemistry; D063646:Carcinogenesis / Q000378:metabolism; D002453:Cell Cycle; D002469:Cell Separation; D002875:Chromosomes / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D056915:DNA Copy Number Variations; D004261:DNA Replication; D004275:DNA, Ribosomal / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D018628:Gene Dosage; D042822:Genomic Instability; D006801:Humans; D006918:Hydroxyurea / Q000737:chemistry; D008164:Luminescent Proteins; D008741:Methyl Methanesulfonate / Q000737:chemistry; D009154:Mutation; D012313:RNA / Q000737:chemistry*; D018741:Replication Origin; D016914:Ribonuclease H / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004275:DNA, Ribosomal; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D012313:RNA; D004247:DNA; D008741:Methyl Methanesulfonate; D016914:Ribonuclease H; D006918:Hydroxyurea; D002166:Camptothecin
keywords: RNA:DNA hybrids; RNase H; replication; ribosomal DNA; topoisomerase 1