title: RPA prevents G-rich structure formation at lagging-strand telomeres to allow maintenance of chromosome ends.
abstract: Replication protein A (RPA) is a highly conserved heterotrimeric single-stranded DNA-binding protein involved in DNA replication, recombination, and repair. In fission yeast, the Rpa1-D223Y mutation provokes telomere shortening. Here, we show that this mutation impairs lagging-strand telomere replication and leads to the accumulation of secondary structures and recruitment of the homologous recombination factor Rad52. The presence of these secondary DNA structures correlates with reduced association of shelterin subunits Pot1 and Ccq1 at telomeres. Strikingly, heterologous expression of the budding yeast Pif1 known to efficiently unwind G-quadruplex rescues all the telomeric defects of the D223Y cells. Furthermore, in vitro data show that the identical D to Y mutation in human RPA specifically affects its ability to bind G-quadruplex. We propose that RPA prevents the formation of G-quadruplex structures at lagging-strand telomeres to promote shelterin association and facilitate telomerase action at telomeres.
authors.affiliation: Cancer Research Center of Marseille (CRCM), U1068 Inserm, UMR7258 CNRS, Institut Paoli-Calmettes, Aix Marseille University (AMU), Marseille, France Ligue Nationale contre le Cancer (LNCC) (Equipe Labellisée), Paris, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Audry
authors.affiliation: Cancer Research Center of Marseille (CRCM), U1068 Inserm, UMR7258 CNRS, Institut Paoli-Calmettes, Aix Marseille University (AMU), Marseille, France Ligue Nationale contre le Cancer (LNCC) (Equipe Labellisée), Paris, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Maestroni
authors.affiliation: Structure des Acides Nucléiques, Télomères et Evolution, Inserm U1154, CNRS UMR 7196, Muséum National d'Histoire Naturelle, Paris Cedex 05, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Delagoutte
authors.affiliation: Laboratoire de Biologie Cellulaire et Moléculaire du Controle de la Prolifération UMR5088, Université de Toulouse, Toulouse, France.
authors.forename: Tiphaine
authors.initials: T
authors.lastname: Gauthier
authors.affiliation: Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, IL, USA.
authors.forename: Toru M
authors.initials: TM
authors.lastname: Nakamura
authors.affiliation: Laboratoire de Biologie Cellulaire et Moléculaire du Controle de la Prolifération UMR5088, Université de Toulouse, Toulouse, France.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Gachet
authors.affiliation: Structure des Acides Nucléiques, Télomères et Evolution, Inserm U1154, CNRS UMR 7196, Muséum National d'Histoire Naturelle, Paris Cedex 05, France UPMC Univ Paris 06, UFR927, Sorbonne Universités, Paris, France.
authors.forename: Carole
authors.initials: C
authors.lastname: Saintomé
authors.affiliation: Cancer Research Center of Marseille (CRCM), U1068 Inserm, UMR7258 CNRS, Institut Paoli-Calmettes, Aix Marseille University (AMU), Marseille, France Ligue Nationale contre le Cancer (LNCC) (Equipe Labellisée), Paris, France vincent.geli@inserm.fr stephane.coulon@inserm.fr.
authors.forename: Vincent
authors.initials: V
authors.lastname: Géli
authors.affiliation: Cancer Research Center of Marseille (CRCM), U1068 Inserm, UMR7258 CNRS, Institut Paoli-Calmettes, Aix Marseille University (AMU), Marseille, France Ligue Nationale contre le Cancer (LNCC) (Equipe Labellisée), Paris, France vincent.geli@inserm.fr stephane.coulon@inserm.fr.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Coulon
mesh_terms: D015825:Chromosomes, Fungal / Q000378:metabolism*; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004256:DNA Polymerase I / Q000378:metabolism; D004257:DNA Polymerase II / Q000378:metabolism; D004261:DNA Replication; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D054856:G-Quadruplexes; D009154:Mutation; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D000089804:Shelterin Complex; D016615:Telomere / Q000737:chemistry / Q000378:metabolism*; D059506:Telomere Shortening; D034501:Telomere-Binding Proteins / Q000378:metabolism
chemical_list: C529647:Ccq1 protein, S pombe; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C497167:RPA1 protein, human; D051716:Replication Protein A; D029702:Schizosaccharomyces pombe Proteins; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins; C499082:pot1 protein, S pombe; C085412:rad52 protein, S pombe; D004256:DNA Polymerase I; D004257:DNA Polymerase II; D004265:DNA Helicases; C468287:Pfh1 protein, S pombe
keywords: G‐quadruplex; RPA; Schizosaccharomyces pombe; replication; telomeres