title: Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery.
abstract: An essential feature of meiosis is Spo11 catalysis of programmed DNA double strand breaks (DSBs). Evidence suggests that the number of DSBs generated per meiosis is genetically determined and that this ability to maintain a pre-determined DSB level, or "DSB homeostasis", might be a property of the meiotic program. Here, we present direct evidence that Rec114, an evolutionarily conserved essential component of the meiotic DSB-machinery, interacts with DSB hotspot DNA, and that Tel1 and Mec1, the budding yeast ATM and ATR, respectively, down-regulate Rec114 upon meiotic DSB formation through phosphorylation. Mimicking constitutive phosphorylation reduces the interaction between Rec114 and DSB hotspot DNA, resulting in a reduction and/or delay in DSB formation. Conversely, a non-phosphorylatable rec114 allele confers a genome-wide increase in both DSB levels and in the interaction between Rec114 and the DSB hotspot DNA. These observations strongly suggest that Tel1 and/or Mec1 phosphorylation of Rec114 following Spo11 catalysis down-regulates DSB formation by limiting the interaction between Rec114 and DSB hotspots. We also present evidence that Ndt80, a meiosis specific transcription factor, contributes to Rec114 degradation, consistent with its requirement for complete cessation of DSB formation. Loss of Rec114 foci from chromatin is associated with homolog synapsis but independent of Ndt80 or Tel1/Mec1 phosphorylation. Taken together, we present evidence for three independent ways of regulating Rec114 activity, which likely contribute to meiotic DSBs-homeostasis in maintaining genetically determined levels of breaks.
authors.affiliation: Department of Life Sciences, Genome Damage and Stability Centre, University of Sussex, Falmer, United Kingdom. jac56@sussex.ac.uk
authors.forename: Jesús A
authors.initials: JA
authors.lastname: Carballo
authors.forename: Silvia
authors.initials: S
authors.lastname: Panizza
authors.forename: Maria Elisabetta
authors.initials: ME
authors.lastname: Serrentino
authors.forename: Anthony L
authors.initials: AL
authors.lastname: Johnson
authors.forename: Marco
authors.initials: M
authors.lastname: Geymonat
authors.forename: Valérie
authors.initials: V
authors.lastname: Borde
authors.forename: Franz
authors.initials: F
authors.lastname: Klein
authors.forename: Rita S
authors.initials: RS
authors.lastname: Cha
mesh_terms: D002843:Chromatin; D053903:DNA Breaks, Double-Stranded; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008540:Meiosis; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D045522:Recombinases / Q000235:genetics* / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D013573:Synaptonemal Complex / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D000099257:Meiotic Recombination Protein SPO11
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D047908:Intracellular Signaling Peptides and Proteins; D017346:Protein Serine-Threonine Kinases; D045522:Recombinases; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D000099257:Meiotic Recombination Protein SPO11; C097071:NDT80 protein, S cerevisiae; C080622:REC114 protein, S cerevisiae; C088937:MEC1 protein, S cerevisiae; C513623:TEL1 protein, S cerevisiae