checkpoint Saccharomyces cerevisiae Rad9 protein contains a tandem tudor domain that recognizes DNA

• Published in: Nucleic acids research 2007-09, Vol.35 (17), p.5898-5912
• Main Authors: Lancelot, Nathalie, Charier, Gaëlle, Couprie, Joël, Duband-Goulet, Isabelle, Alpha-Bazin, Béatrice, Quémeneur, Eric, Ma, Emilie, Marsolier-Kergoat, Marie-Claude, Ropars, Virginie, Charbonnier, Jean-Baptiste, Miron, Simona, Craescu, Constantin T, Callebaut, Isabelle, Gilquin, Bernard, Zinn-Justin, Sophie
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Biochemistry, Molecular Biology; Cell Cycle Proteins - chemistry; Cell Cycle Proteins - metabolism; Checkpoint Kinase 2; DNA - chemistry; DNA - metabolism; DNA Breaks, Double-Stranded; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - metabolism; G1 Phase; Histones - chemistry; Histones - metabolism; Humans; Intracellular Signaling Peptides and Proteins - chemistry; Life Sciences; Metazoa; Mice; Molecular Sequence Data; Nuclear Proteins - chemistry; Protein Structure, Tertiary; Protein-Serine-Threonine Kinases - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - radiation effects; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - metabolism; Schizosaccharomyces pombe; Schizosaccharomyces pombe Proteins - chemistry; Sequence Alignment; Structural Biology; Tumor Suppressor p53-Binding Protein 1
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkm607

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.