The G2 checkpoint activated by DNA damage does not prevent genome instability in plant cells

Root growth, G2 length, and the frequency of aberrant mitoses and apoptotic nuclei were recorded after a single X-ray irradiation, ranging from 2.5 to 40 Gy, in Allium cepa L. root meristematic cells. After 72 h of recovery, root growth was reduced in a dose-dependent manner from 10 to 40 Gy, but not at 2.5 or 5 Gy doses. Flow cytometry plus TUNEL (TdT-mediated dUTP nick end labeling) showed that activation of apoptosis occurred only after 20 and 40 Gy of X-rays. Nevertheless, irrespective of the radiation dose, conventional flow cytometry showed that cells accumulated in G2 (4C DNA content). Simultaneously, the mitotic index fell, though a mitotic wave appeared later. Cell accumulation in G2 was transient and partially reversed by caffeine, thus it was checkpoint-dependent. Strikingly, the additional G2 time provided by this checkpoint was never long enough to complete DNA repair. Then, in all cases, some G2 cells with still-unrepaired DNA underwent checkpoint adaptation, i.e., they entered into the late mitotic wave with chromatid breaks. These cells and those produced by the breakage of chromosomal bridges in anaphase will reach the G1 of the next cell cycle unrepaired, ensuring the appearance of genome instabilit

Transcription of ribosomal genes can cause nondisjunction

Mitotic disjunction of the repetitive ribosomal DNA (rDNA) involves specialized segregation mechanisms dependent on the conserved phosphatase Cdc14. The reason behind this requirement is unknown. We show that rDNA segregation requires Cdc14 partly because of its physical length but most importantly because a fraction of ribosomal RNA (rRNA) genes are transcribed at very high rates. We show that cells cannot segregate rDNA without Cdc14 unless they undergo genetic rearrangements that reduce rDNA copy number. We then demonstrate that cells with normal length rDNA arrays can segregate rDNA in the absence of Cdc14 as long as rRNA genes are not transcribed. In addition, our study uncovers an unexpected role for the replication barrier protein Fob1 in rDNA segregation that is independent of Cdc14. These findings demonstrate that highly transcribed loci can cause chromosome nondisjunction

A revised view of the Canis Major stellar overdensity with DECam and Gaia: new evidence of a stellar warp of blue stars

We present DECam imaging combined with Gaia DR2 data to study the Canis Major overdensity. The presence of the so-called Blue Plume stars in a low-pollution area of the color-magnitude diagram allows us to derive the distance and proper motions of this stellar feature along the line of sight of its hypothetical core. The stellar overdensity extends on a large area of the sky at low Galactic latitudes, below the plane, and between 230$^\circ < \ell < 255^\circ$. According to the orbit derived for Canis Major, it presents an on-plane rotation around the Milky Way. Moreover, additional overdensities of Blue Plume stars are found around the plane and across the Galaxy, proving that these objects are not only associated with that structure. The spatial distribution of these stars, derived using Gaia astrometric data, confirms that the detection of the Canis Major overdensity results more from the warped structure of the Milky Way disk than from the accretion of a dwarf galaxy.Comment: 11 pages, 9 figures, accepted for publication in MNRA

Essential and checkpoint functions of budding yeast ATM and ATR during meiotic prophase are facilitated by differential phosphorylation of a meiotic adaptor protein, Hop1

A hallmark of the conserved ATM/ATR signalling is its ability to mediate a wide range of functions utilizing only a limited number of adaptors and effector kinases. During meiosis, Tel1 and Mec1, the budding yeast ATM and ATR, respectively, rely on a meiotic adaptor protein Hop1, a 53BP1/Rad9 functional analog, and its associated kinase Mek1, a CHK2/Rad53-paralog, to mediate multiple functions: control of the formation and repair of programmed meiotic DNA double strand breaks, enforcement of inter-homolog bias, regulation of meiotic progression, and implementation of checkpoint responses. Here, we present evidence that the multi-functionality of the Tel1/Mec1-to-Hop1/Mek1 signalling depends on stepwise activation of Mek1 that is mediated by Tel1/Mec1 phosphorylation of two specific residues within Hop1: phosphorylation at the threonine 318 (T318) ensures the transient basal level Mek1 activation required for viable spore formation during unperturbed meiosis. Phosphorylation at the serine 298 (S298) promotes stable Hop1-Mek1 interaction on chromosomes following the initial phospho-T318 mediated Mek1 recruitment. In the absence of Dmc1, the phospho-S298 also promotes Mek1 hyper-activation necessary for implementing meiotic checkpoint arrest. Taking these observations together, we propose that the Hop1 phospho-T318 and phospho-S298 constitute key components of the Tel1/Mec1- based meiotic recombination surveillance (MRS) network and facilitate effective coupling of meiotic recombination and progression during both unperturbed and challenged meiosis

Stellar streams around the Magellanic Clouds in 4D

We carried out a spectroscopic follow-up program of the four new stellar stream candidates detected by Belokurov & Koposov (2016) in the outskirts of the Large Magellanic Cloud (LMC) using FORS2 (VLT). The medium-resolution spectra were used to measure the line-of-sight velocities, estimate stellar metallicities and to classify stars into Blue Horizontal Branch (BHB) and Blue Straggler (BS) stars. Using the 4-D phase-space information, we attribute approximately one half of our sample to the Magellanic Clouds, while the rest is part of the Galactic foreground. Only two of the four stream candidates are confirmed kinematically. While it is impossible to estimate the exact levels of MW contamination, the phase-space distribution of the entire sample of our Magellanic stars matches the expected velocity gradient for the LMC halo and extends as far as 33 deg (angular separation) or 29 kpc from the LMC center. Our detections reinforce the idea that the halo of the LMC seems to be larger than previously expected, and its debris can be spread in the sky out to very large separations from the LMC center. Finally, we provide some kinematic evidence that many of the stars analysed here have likely come from the Small Magellanic Cloud.Comment: 15 pages, 13 figures, accepted for publication in MNRA

Effect of pH on the production of hydrolytic enzymes of industrial interest by Bacillus licheniformis using sewage sludge as a low-cost culture médium

Management of sewage sludge has become a topic of increasing importance. This is because the treatment and final destination of these wastes are not growing at the same rate at which generated what is necessary to add the limited storage capacity of wastewater treatment plants or waste transfer centers. Current policies of different governments aims, among others, promote proecological management of sewage sludge, replacing traditional methods by new methods that lead to their stabilization and safe recycling. These methods can lead to the recovery of valuable raw materials from potentially dangerous others in order to permit their use in agriculture, industry or energy production. Moreover, exists a great enzyme market, mainly those with hydrolytic activities such as proteases, lipases or cellulases. Although its use has many advantages, the competitiveness of the enzymes compared with chemicals is limited by its high production costs. The use of a less expensive culture media and the stimulation of the production of these enzymes can significantly reduce production costs. Therefore, the use of tertiary materials as alternative substrates, available in large quantities and at lower cost, is an interesting option. In this work we have studied the effect of pH on the fermentation of sewage sludge by Bacillus licheniformis, a hydrolytic enzymes producer microorganism, in order to obtain several products for specific agronomic applications

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