Budding yeast ATM/ATR control meiotic double-strand break (DSB) levels by down-regulating Rec114, an essential component of the DSB-machinery

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

Budding Yeast Pch2, a Widely Conserved Meiotic Protein, Is Involved in the Initiation of Meiotic Recombination

Budding yeast Pch2 protein is a widely conserved meiosis-specific protein whose role is implicated in the control of formation and displacement of meiotic crossover events. In contrast to previous studies where the function of Pch2 was implicated in the steps after meiotic double-strand breaks (DSBs) are formed, we present evidence that Pch2 is involved in meiotic DSB formation, the initiation step of meiotic recombination. The reduction of DSB formation caused by the pch2 mutation is most prominent in the sae2 mutant background, whereas the impact remains mild in the rad51 dmc1 double mutant background. The DSB reduction is further pronounced when pch2 is combined with a hypomorphic allele of SPO11. Interestingly, the level of DSB reduction is highly variable between chromosomes, with minimal impact on small chromosomes VI and III. We propose a model in which Pch2 ensures efficient formation of meiotic DSBs which is necessary for igniting the subsequent meiotic checkpoint responses that lead to proper differentiation of meiotic recombinants

Physical properties of Centaur (60558) 174P/Echeclus from stellar occultations

The Centaur (60558) Echeclus was discovered on March 03, 2000, orbiting between the orbits of Jupiter and Uranus. After exhibiting frequent outbursts, it also received a comet designation, 174P. If the ejected material can be a source of debris to form additional structures, studying the surroundings of an active body like Echeclus can provide clues about the formation scenarios of rings, jets, or dusty shells around small bodies. Stellar occultation is a handy technique for this kind of investigation, as it can, from Earth-based observations, detect small structures with low opacity around these objects. Stellar occultation by Echeclus was predicted and observed in 2019, 2020, and 2021. We obtain upper detection limits of rings with widths larger than 0.5 km and optical depth of $\tau$ = 0.02. These values are smaller than those of Chariklo's main ring; in other words, a Chariklo-like ring would have been detected. The occultation observed in 2020 provided two positive chords used to derive the triaxial dimensions of Echeclus based on a 3D model and pole orientation available in the literature. We obtained $a = 37.0\pm0.6$ km, $b = 28.4 \pm 0.5$ km, and $c= 24.9 \pm 0.4$ km, resulting in an area-equivalent radius of $30.0 \pm 0.5$ km. Using the projected limb at the occultation epoch and the available absolute magnitude ($\rm{H}_{\rm{v}} = 9.971 \pm 0.031$), we calculate an albedo of $p_{\rm{v}} = 0.050 \pm 0.003$. Constraints on the object's density and internal friction are also proposed.Comment: Corrected and typeset versio

A High Throughput Genetic Screen Identifies New Early Meiotic Recombination Functions in Arabidopsis thaliana

Meiotic recombination is initiated by the formation of numerous DNA double-strand breaks (DSBs) catalysed by the widely conserved Spo11 protein. In Saccharomyces cerevisiae, Spo11 requires nine other proteins for meiotic DSB formation; however, unlike Spo11, few of these are conserved across kingdoms. In order to investigate this recombination step in higher eukaryotes, we took advantage of a high-throughput meiotic mutant screen carried out in the model plant Arabidopsis thaliana. A collection of 55,000 mutant lines was screened, and spo11-like mutations, characterised by a drastic decrease in chiasma formation at metaphase I associated with an absence of synapsis at prophase, were selected. This screen led to the identification of two populations of mutants classified according to their recombination defects: mutants that repair meiotic DSBs using the sister chromatid such as Atdmc1 or mutants that are unable to make DSBs like Atspo11-1. We found that in Arabidopsis thaliana at least four proteins are necessary for driving meiotic DSB repair via the homologous chromosomes. These include the previously characterised DMC1 and the Hop1-related ASY1 proteins, but also the meiotic specific cyclin SDS as well as the Hop2 Arabidopsis homologue AHP2. Analysing the mutants defective in DSB formation, we identified the previously characterised AtSPO11-1, AtSPO11-2, and AtPRD1 as well as two new genes, AtPRD2 and AtPRD3. Our data thus increase the number of proteins necessary for DSB formation in Arabidopsis thaliana to five. Unlike SPO11 and (to a minor extent) PRD1, these two new proteins are poorly conserved among species, suggesting that the DSB formation mechanism, but not its regulation, is conserved among eukaryotes

Physical properties of Centaur (60558) 174P/Echeclus from stellar occultations

peer reviewedThe Centaur (60558) Echeclus was discovered on 2000 March 03, orbiting between the orbits of Jupiter and Uranus. After exhibiting frequent outbursts, it also received a comet designation, 174P. If the ejected material can be a source of debris to form additional structures, studying the surroundings of an active body like Echeclus can provide clues about the formation scenarios of rings, jets, or dusty shells around small bodies. Stellar occultation is a handy technique for this kind of investigation, as it can, from Earth-based observations, detect small structures with low opacity around these objects. Stellar occultation by Echeclus was predicted and observed in 2019, 2020, and 2021. We obtain upper detection limits of rings with widths larger than 0.5 km and optical depth of τ = 0.02. These values are smaller than those of Chariklo's main ring; in other words, a Chariklo-like ring would have been detected. The occultation observed in 2020 provided two positive chords used to derive the triaxial dimensions of Echeclus based on a 3D model and pole orientation available in the literature. We obtained a = 37.0 ± 0.6 km, b = 28.4 ± 0.5 km, and c = 24.9 ± 0.4 km, resulting in an area-equivalent radius of 30.0 ± 0.5 km. Using the projected limb at the occultation epoch and the available absolute magnitude (Hv= 9.971 +- 0.031), we calculate an albedo of pv = 0.050 ± 0.003. Constraints on the object's density and internal friction are also proposed

Break-Induced Replication Requires DNA Damage-Induced Phosphorylation of Pif1 and Leads to Telomere Lengthening

Broken replication forks result in DNA breaks that are normally repaired via homologous recombination or break induced replication (BIR). Mild insufficiency in the replicative ligase Cdc9 in budding yeast Saccharomyces cerevisiae resulted in a population of cells with persistent DNA damage, most likely due to broken replication forks, constitutive activation of the DNA damage checkpoint and longer telomeres. This telomere lengthening required functional telomerase, the core DNA damage signaling cascade Mec1-Rad9-Rad53, and the components of the BIR repair pathway - Rad51, Rad52, Pol32, and Pif1. The Mec1-Rad53 induced phosphorylation of Pif1, previously found necessary for inhibition of telomerase at double strand breaks, was also important for the role of Pif1 in BIR and telomere elongation in cdc9-1 cells. Two other mutants with impaired DNA replication, cdc44-5 and rrm3Δ, were similar to cdc9-1: their long telomere phenotype was dependent on the Pif1 phosphorylation locus. We propose a model whereby the passage of BIR forks through telomeres promotes telomerase activity and leads to telomere lengthening