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Mutations in the Saccharomyces cerevisiae CDC1 gene affect double-strand-break-induced intrachromosomal recombination. 总被引:1,自引:0,他引:1
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To isolate Saccharomyces cerevisiae mutants defective in recombinational DNA repair, we constructed a strain that contains duplicated ura3 alleles that flank LEU2 and ADE5 genes at the ura3 locus on chromosome V. When a HO endonuclease cleavage site is located within one of the ura3 alleles, Ura+ recombination is increased over 100-fold in wild-type strains following HO induction from the GAL1, 10 promoter. This strain was used to screen for mutants that exhibited reduced levels of HO-induced intrachromosomal recombination without significantly affecting the spontaneous frequency of Ura+ recombination. One of the mutations isolated through this screen was found to affect the essential gene CDC1. This mutation, cdc1-100, completely eliminated HO-induced Ura+ recombination yet maintained both spontaneous preinduced recombination levels and cell viability, cdc1-100 mutants were moderately sensitive to killing by methyl methanesulfonate and gamma irradiation. The effect of the cdc1-100 mutation on recombinational double-strand break repair indicates that a recombinationally silent mechanism other than sister chromatid exchange was responsible for the efficient repair of DNA double-strand breaks. 相似文献
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DNA-dependent protein kinase (DNA-PK), composed of Ku70, Ku80, and the catalytic subunit (DNA-PKcs), is involved in double-strand break (DSB) repair by non-homologous end joining (NHEJ). DNA-PKcs defects confer ionizing radiation sensitivity and increase homologous recombination (HR). Increased HR is consistent with passive shunting of DSBs from NHEJ to HR. We therefore predicted that inhibiting the DNA-PKcs kinase would increase HR. A novel DNA-PKcs inhibitor (1-(2-hydroxy-4-morpholin-4-yl-phenyl)-ethanone; designated IC86621) increased ionizing radiation sensitivity but surprisingly decreased spontaneous and DSB-induced HR. Wortmannin also inhibits DNA-PKcs and reduces DSB-induced HR. IC86621 did not affect HR product outcome, indicating that it affects HR initiation. Thus, HR is increased in the absence of DNA-PKcs, but decreased when DNA-PKcs is catalytically inactive, suggesting interactive competition between HR and NHEJ. The effects of IC86621 and wortmannin were proportional to the level of DNA-PKcs, consistent with inhibited DNA-PKcs acting in a dominant negative manner. We propose that inhibition of DNA-PKcs blocks its autophosphorylation, prevents dissociation of DNA-PKcs from DNA ends, and thereby blocks both HR and NHEJ. By blocking the two major DSB repair pathways, DNA-PKcs inhibitors should radiosensitize at all cell-cycle stages and are therefore excellent candidates for augmenting cancer radiotherapy. 相似文献
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Stephanie C. Plautz Richard S. Halbrook Donald W. Sparling 《The Journal of wildlife management》2011,75(4):779-785
Previous field studies of hunter-harvested mourning doves (Zenaida macroura) have reported the percentage of birds with ingested lead shot as 0.2–6.5%. To reduce the uncertainty concerning the number of doves that ingest shot, we conducted an experiment to test the proportion of mourning doves that ingested lead shot on the bare soil of a disked field (typical of a managed dove field) to simulate more natural feeding conditions. In each of 3 treatment groups of 80 birds, we exposed 35 birds to low-density lead shot (1.5 million shot/ha), and35 birds to high-density lead shot (29.5 million shot/ha), and 10 birds served as controls (no shot). We dosed 5 positive control birds with 2 lead shot each in trials 2 and 3. We scattered lead shot and mixed seed on the loosely packed soil of treatment cages and after 4 days of exposure, 2.9% of doves voluntarily ingested ≥1 lead shot. The proportion of birds that ingested shot when exposed to the high-density shot treatment (4.9%) was not different (P = 0.098) from that of the low-density shot treatment (1.0%). Lead concentrations in liver, kidneys, and blood reached maxima of 94.402 ppm, 346.033 ppm, and 13.883 ppm wet mass, respectively. Differences in delta-aminolevulinic acid dehydratase (ALAD) activity, packed cell volume, and heterophil:lymphocyte ratio (H:L) were greater posttreatment in doves that had ingested shot than in those that did not. The risk posed to mourning doves from lead shot ingestion can be reduced by banning lead shot on management areas or dove fields or disking fields after hunting season to reduce shot availability. © 2011 The Wildlife Society. 相似文献
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Anna Bratcher Nicole A. Hoff Reena H. Doshi Adva Gadoth Megan Halbrook Patrick Mukadi Kamy Musene Benoit Ilunga-Kebela DAndre Spencer Matthew S. Bramble David McIlwan J. Daniel Kelly Daniel Mukadi Placide Mbala Kingebeni Steve Ahuka Emile Okitolonda-Wemakoy Jean-Jacques Muyembe-Tamfum Anne W. Rimoin 《PLoS neglected tropical diseases》2021,15(8)
BackgroundEbola virus (EBOV) is a zoonotic filovirus spread through exposure to infected bodily fluids of a human or animal. Though EBOV is capable of causing severe disease, referred to as Ebola Virus Disease (EVD), individuals who have never been diagnosed with confirmed, probable or suspected EVD can have detectable EBOV antigen-specific antibodies in their blood. This study aims to identify risk factors associated with detectable antibody levels in the absence of an EVD diagnosis.MethodologyData was collected from September 2015 to August 2017 from 1,366 consenting individuals across four study sites in the DRC (Boende, Kabondo-Dianda, Kikwit, and Yambuku). Seroreactivity was determined to EBOV GP IgG using Zaire Ebola Virus Glycoprotein (EBOV GP antigen) ELISA kits (Alpha Diagnostic International, Inc.) in Kinshasa, DRC; any result above 4.7 units/mL was considered seroreactive. Among the respondents, 113 (8.3%) were considered seroreactive. Several zoonotic exposures were associated with EBOV seroreactivity after controlling for age, sex, healthcare worker status, location, and history of contact with an EVD case, namely: ever having contact with bats, ever having contact with rodents, and ever eating non-human primate meat. Contact with monkeys or non-human primates was not associated with seroreactivity.ConclusionsThis analysis suggests that some zoonotic exposures that have been linked to EVD outbreaks can also be associated with EBOV GP seroreactivity in the absence of diagnosed EVD. Future investigations should seek to clarify the relationships between zoonotic exposures, seroreactivity, asymptomatic infection, and EVD. 相似文献
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Abigail S. Krall Peter J. Mullen Felicia Surjono Milica Momcilovic Ernst W. Schmid Christopher J. Halbrook Apisadaporn Thambundit Steven D. Mittelman Costas A. Lyssiotis David B. Shackelford Simon R.V. Knott Heather R. Christofk 《Cell metabolism》2021,33(5):1013-1026.e6
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