title: The checkpoint Saccharomyces cerevisiae Rad9 protein contains a tandem tudor domain that recognizes DNA.
abstract: DNA damage checkpoints are signal transduction pathways that are activated after genotoxic insults to protect genomic integrity. At the site of DNA damage, 'mediator' proteins are in charge of recruiting 'signal transducers' to molecules 'sensing' the damage. Budding yeast Rad9, fission yeast Crb2 and metazoan 53BP1 are presented as mediators involved in the activation of checkpoint kinases. Here we show that, despite low sequence conservation, Rad9 exhibits a tandem tudor domain structurally close to those found in human/mouse 53BP1 and fission yeast Crb2. Moreover, this region is important for the resistance of Saccharomyces cerevisiae to different genotoxic stresses. It does not mediate direct binding to a histone H3 peptide dimethylated on K79, nor to a histone H4 peptide dimethylated on lysine 20, as was demonstrated for 53BP1. However, the tandem tudor region of Rad9 directly interacts with single-stranded DNA and double-stranded DNAs of various lengths and sequences through a positively charged region absent from 53BP1 and Crb2 but present in several yeast Rad9 homologs. Our results argue that the tandem tudor domains of Rad9, Crb2 and 53BP1 mediate chromatin binding next to double-strand breaks. However, their modes of chromatin recognition are different, suggesting that the corresponding interactions are differently regulated.
authors.affiliation: Institut de Biologie et Technologies de Saclay, CEA Saclay, 91191 Gif-sur-Yvette, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Lancelot
authors.forename: Gaëlle
authors.initials: G
authors.lastname: Charier
authors.forename: Joël
authors.initials: J
authors.lastname: Couprie
authors.forename: Isabelle
authors.initials: I
authors.lastname: Duband-Goulet
authors.forename: Béatrice
authors.initials: B
authors.lastname: Alpha-Bazin
authors.forename: Eric
authors.initials: E
authors.lastname: Quémeneur
authors.forename: Emilie
authors.initials: E
authors.lastname: Ma
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: Marsolier-Kergoat
authors.forename: Virginie
authors.initials: V
authors.lastname: Ropars
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Charbonnier
authors.forename: Simona
authors.initials: S
authors.lastname: Miron
authors.forename: Constantin T
authors.initials: CT
authors.lastname: Craescu
authors.forename: Isabelle
authors.initials: I
authors.lastname: Callebaut
authors.forename: Bernard
authors.initials: B
authors.lastname: Gilquin
authors.forename: Sophie
authors.initials: S
authors.lastname: Zinn-Justin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism; D064447:Checkpoint Kinase 2; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D016193:G1 Phase; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry; D016415:Sequence Alignment; D000071857:Tumor Suppressor p53-Binding Protein 1
chemical_list: D018797:Cell Cycle Proteins; C109867:Crb2 protein, S pombe; D004268:DNA-Binding Proteins; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; C498301:TP53BP1 protein, human; D000071857:Tumor Suppressor p53-Binding Protein 1; C069930:rad9 protein; D004247:DNA; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae