Whole genome RNAi screens reveal a critical role of REV3 in coping with replication stress

REV3, the catalytic subunit of translesion polymerase zeta (polζ), is commonly associated with DNA damage bypass and repair. Despite sharing accessory subunits with replicative polymerase δ, very little is known about the role of polζ in DNA replication. We previously demonstrated that inhibition of REV3 expression induces persistent DNA damage and growth arrest in cancer cells. To reveal determinants of this sensitivity and obtain insights into the cellular function of REV3, we performed whole human genome RNAi library screens aimed at identification of synthetic lethal interactions with REV3 in A549 lung cancer cells. The top confirmed hit was RRM1, the large subunit of ribonucleotide reductase (RNR), a critical enzyme of de novo nucleotide synthesis. Treatment with the RNR-inhibitor hydroxyurea (HU) synergistically increased the fraction of REV3-deficient cells containing single stranded DNA (ssDNA) as indicated by an increase in replication protein A (RPA). However, this increase was not accompanied by accumulation of the DNA damage marker γH2AX suggesting a role of REV3 in counteracting HU-induced replication stress (RS). Consistent with a role of REV3 in DNA replication, increased RPA staining was confined to HU-treated S-phase cells. Additionally, we found genes related to RS to be significantly enriched among the top hits of the synthetic sickness/lethality (SSL) screen further corroborating the importance of REV3 for DNA replication under conditions of RS

Short-Term Oxidation of HfB₂-SiC Based UHTC in Supersonic Flow of Carbon Dioxide Plasma

The short-term (5 min) exposure to the supersonic flow of carbon dioxide plasma on ultrahigh-temperature ceramics of HfB2-30vol.%SiC composition has been studied. It was shown that, when established on the surface at a temperature of 1615–1655 °C, the beginning of the formation of an oxidized layer takes place. Raman spectroscopy and scanning electron microscopy studies showed that the formation of a porous SiC-depleted region is not possible under the HfO2-SiO2 surface oxide layer. Numerical modeling based on the Navier–Stokes equations and experimental probe measurements of the test conditions were performed. The desirability of continuing systematic studies on the behavior of ultrahigh-temperature ZrB2/HfB2-SiC ceramics, including those doped with various components under the influence of high-enthalpy gas flows, was noted

Diagnostic Mirrors for ITER: Research in the Frame of International Tokamak Physics Activity

Mirrors will be used as first plasma-viewing elements in optical and laser-based diagnostics in ITER. Deterioration of the mirror performance due to e.g. sputtering of the mirror surface by plasma particles or deposition of impurities will hamper the entire performance of the affected diagnostic and thus affect ITER operation. The Specialists Working Group on First Mirrors (FM SWG) in the Topical Group on Diagnostics of the International Tokamak Physics Activity (ITPA) plays an important role in finding solutions for diagnostic first mirrors. Sound progress in research and development of diagnostic mirrors in ITER was achieved since the last overview in 2009. Single crystal (SC) rhodium (Rh) mirrors became available. SC rhodium and molybdenum (Mo) mirrors survived in conditions corresponding to ~200 cleaning cycles with a negligible degradation of reflectivity. These results are important for a mirror cleaning system which is presently under development. The cleaning system is based on sputtering of contaminants by plasma. Repetitive cleaning was tested on several mirror materials. Experiments comprised contamination/cleaning cycles. The reflectivity SC Mo and Rh mirrors has changed insignificantly after 80 cycles. First in situ cleaning using radiofrequency (RF) plasma was conducted in EAST tokamak with a mock-up plate of ITER edge Thomson Scattering (ETS) with five inserted mirrors. Contaminants from the mirrors were removed. Physics of cleaning discharge was studied both experimentally and by modeling. Mirror contamination can also be mitigated by protecting diagnostic ducts. A deposition mitigation (DeMi) duct system was exposed in KSTAR. The real-time measurement of deposition in the diagnostic duct was pioneered during this experiment. Results evidenced the dominating effect of the wall conditioning and baking on contamination inside the duct. A baffled cassette with mirrors was exposed at the main wall of JET for 23,6 plasma hours. No significant degradation of reflectivity was measured on mirrors located in the ducts. Predictive modeling was further advanced. A model for the particle transport, deposition and erosion at the port-plug was used in selecting an optical layout of several ITER diagnostics. These achievements contributed to the focusing of the first mirror research thus accelerating the diagnostic development. Modeling requires more efforts. Remaining crucial issues will be in a focus of the future work of the FM SWG

Inhibition of REV3 Expression Induces Persistent DNA Damage and Growth Arrest in Cancer Cells12

REV3 is the catalytic subunit of DNA translesion synthesis polymerase ζ. Inhibition of REV3 expression increases the sensitivity of human cells to a variety of DNA-damaging agents and reduces the formation of resistant cells. Surprisingly, we found that short hairpin RNA-mediated depletion of REV3 per se suppresses colony formation of lung (A549, Calu-3), breast (MCF-7, MDA-MB-231), mesothelioma (IL45 and ZL55), and colon (HCT116 +/-p53) tumor cell lines, whereas control cell lines (AD293, LP9-hTERT) and the normal mesothelial primary culture (SDM104) are less affected. Inhibition of REV3 expression in cancer cells leads to an accumulation of persistent DNA damage as indicated by an increase in phospho-ATM, 53BP1, and phospho-H2AX foci formation, subsequently leading to the activation of the ATM-dependent DNA damage response cascade. REV3 depletion in p53-proficient cancer cell lines results in a G1 arrest and induction of senescence as indicated by the accumulation of p21 and an increase in senescence-associated β-galactosidase activity. In contrast, inhibition of REV3 expression in p53-deficient cells results in growth inhibition and a G2/M arrest. A small fraction of the p53-deficient cancer cells can overcome the G2/M arrest, which results in mitotic slippage and aneuploidy. Our findings reveal that REV3 depletion per se suppresses growth of cancer cell lines from different origin, whereas control cell lines and a mesothelial primary culture were less affected. Thus, our findings indicate that depletion of REV3 not only can amend cisplatin-based cancer therapy but also can be applied for susceptible cancers as a potential monotherapy

Energy calibration and resolution of the CMS electromagnetic calorimeter in pp collisions at s\sqrt{s} = 7 TeV

The energy calibration and resolution of the electromagnetic calorimeter (ECAL) of the CMS detector have been determined using proton-proton collision data from LHC operation in 2010 and 2011 at a centre-of-mass energy of sqrt(s)=7 TeV with integrated luminosities of about 5 inverse femtobarns. Crucial aspects of detector operation, such as the environmental stability, alignment, and synchronization, are presented. The in-situ calibration procedures are discussed in detail and include the maintenance of the calibration in the challenging radiation environment inside the CMS detector. The energy resolution for electrons from Z-boson decays is better than 2% in the central region of the ECAL barrel (for pseudorapidity abs(eta)<0.8) and is 2-5% elsewhere. The derived energy resolution for photons from 125 GeV Higgs boson decays varies across the barrel from 1.1% to 2.6% and from 2.2% to 5% in the entraps. The calibration of the absolute energy is determined from Z to e+e- decays to a precision of 0.4% in the barrel and 0.8% in the endcaps

Search for long-lived particles in events with photons and missing energy in proton\u2013proton collisions at sqrt(s)=7 TeV

Results are presented from a search for long-lived neutralinos decaying into a photon and an invisible particle, a signature associated with gauge-mediated supersymmetry breaking in supersymmetric models. The analysis is based on a 4.9 inverse femtobarn sample of proton-proton collisions at 1as = 7 TeV, collected with the CMS detector at the LHC. The missing transverse energy and the time of arrival of the photon at the electromagnetic calorimeter are used to search for an excess of events over the expected background. No significant excess is observed, and lower limits at the 95% confidence level are obtained on the mass of the lightest neutralino, m(neutralino) > 220 GeV (for c tau 6000 mm (for m(neutralino) < 150 GeV)