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Iben B. Bentsen Ida Nielsen Michael Lisby Helena B. Nielsen Souvik Sen Gupta Kamilla Mundbjerg Anni H. Andersen Lotte Bjergbaek 《Nucleic acids research》2013,41(5):3173-3189
To address how eukaryotic replication forks respond to fork stalling caused by strong non-covalent protein–DNA barriers, we engineered the controllable Fob-block system in Saccharomyces cerevisiae. This system allows us to strongly induce and control replication fork barriers (RFB) at their natural location within the rDNA. We discover a pivotal role for the MRX (Mre11, Rad50, Xrs2) complex for fork integrity at RFBs, which differs from its acknowledged function in double-strand break processing. Consequently, in the absence of the MRX complex, single-stranded DNA (ssDNA) accumulates at the rDNA. Based on this, we propose a model where the MRX complex specifically protects stalled forks at protein–DNA barriers, and its absence leads to processing resulting in ssDNA. To our surprise, this ssDNA does not trigger a checkpoint response. Intriguingly, however, placing RFBs ectopically on chromosome VI provokes a strong Rad53 checkpoint activation in the absence of Mre11. We demonstrate that proper checkpoint signalling within the rDNA is restored on deletion of SIR2. This suggests the surprising and novel concept that chromatin is an important player in checkpoint signalling. 相似文献
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A wide-ranging examination of plastid (pt)DNA sequence homologies within
higher plant nuclear genomes (promiscuous DNA) was undertaken. Digestion
with methylation-sensitive restriction enzymes and Southern analysis was
used to distinguish plastid and nuclear DNA in order to assess the extent
of variability of promiscuous sequences within and between plant species.
Some species, such as Gossypium hirsutum (cotton), Nicotiana tabacum
(tobacco), and Chenopodium quinoa, showed homogenity of these sequences,
while intraspecific sequence variation was observed among different
cultivars of Pisum sativum (pea), Hordeum vulgare (barley), and Triticum
aestivum (wheat). Hypervariability of plastid sequence homologies was
identified in the nuclear genomes of Spinacea oleracea (spinach) and Beta
vulgaris (beet), in which individual plants were shown to possess a unique
spectrum of nuclear sequences with ptDNA homology. This hypervariability
apparently extended to somatic variation in B. vulgaris. No sequences with
ptDNA homology were identified by this method in the nuclear genome of
Arabidopsis thaliana.
相似文献
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A survey was conducted to determine the levels of fumonisins B1 and B2 in corn and corn-based products available in Colombia
for human and animal consumption. A total of 120 samples were analyzed by acetonitrile-water extraction, cleanup with a strong-anion-exchange
column, and liquid chromatography with o-phthaldialdehyde-2-mercaptoethanol derivatization and fluorescence detection. The
samples of corn and corn-based products for animal intake were taken at different feed manufacturing plants, whereas the samples
used for human foods where purchased from local retail stores. The number of positive samples for fumonisin B1 was 20.0% higher
in corn and corn-based products for animal intake (75.0%) than in corn and corn-based products for human consumption (55.0%).
The levels of fumonisin B1 were also higher in corn and corn-based products for animal intake (mean = 694 μg/kg; range = 32–2964
μg/kg), than in corn and corn-based products for human intake (mean = 218 μg/kg; range = 24–2170 μg/ kg). The incidence and
levels of fumonisin B2 were lower than those for fumonisin B1. Corn and corn-based products for animal consumption had an
incidence of fumonisin B2 of 58.3%, with a mean value of 283 μg/kg, and a range of 44–987 μg/kg. The incidence of fumonisin
B2 in corn-based products for human intake was 35.0%, with a mean value of 118 μg/kg and a range of 21–833 μg/kg. The highest
incidence and levels of fumonisins were found in samples of hominy feed, with concentrations ranging from 86 to 2964 μg/kg
fumonisin B1 and 57 to 987 μg/kg fumonisin B2. 相似文献
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Kamilla Mundbjerg Signe W. J?rgensen Jacob Freds?e Ida Nielsen Jakob Madsen Pedersen Iben Bach Bentsen Michael Lisby Lotte Bjergbaek Anni H Andersen 《PLoS genetics》2015,11(12)
Faithful DNA replication with correct termination is essential for genome stability and transmission of genetic information. Here we have investigated the potential roles of Topoisomerase II (Top2) and the RecQ helicase Sgs1 during late stages of replication. We find that cells lacking Top2 and Sgs1 (or Top3) display two different characteristics during late S/G2 phase, checkpoint activation and accumulation of asymmetric X-structures, which are both independent of homologous recombination. Our data demonstrate that checkpoint activation is caused by a DNA structure formed at the strongest rDNA replication fork barrier (RFB) during replication termination, and consistently, checkpoint activation is dependent on the RFB binding protein, Fob1. In contrast, asymmetric X-structures are formed independent of Fob1 at less strong rDNA replication fork barriers. However, both checkpoint activation and formation of asymmetric X-structures are sensitive to conditions, which facilitate fork merging and progression of replication forks through replication fork barriers. Our data are consistent with a redundant role of Top2 and Sgs1 together with Top3 (Sgs1-Top3) in replication fork merging at rDNA barriers. At RFB either Top2 or Sgs1-Top3 is essential to prevent formation of a checkpoint activating DNA structure during termination, but at less strong rDNA barriers absence of the enzymes merely delays replication fork merging, causing an accumulation of asymmetric termination structures, which are solved over time. 相似文献
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Jette Junge Kirsten D Bentsen Per Christoffersen Marianne Orholm Thorkild I A S?rensen Thomas Horn 《BMJ (Clinical research ed.)》1988,296(6637):1629-1630
In a study of 142 male alcohol abusers without evidence of cirrhosis the presence of intralobular fibronectin in the liver was investigated in relation to the subsequent development of the disease. All 142 initial biopsy samples showed preserved architecture. During a follow up period of 10-13 years 23 patients (16%) developed cirrhosis. Twelve of 110 patients with normal or slightly increased amounts of parenchymal fibronectin in the initial biopsy specimen developed cirrhosis, whereas eight out of 27 patients with moderately increased amounts and three out of five with significantly increased amounts later developed the disease (p<0·005).Semiquantitative assessment of the amount of parenchymal fibronectin at an early stage of alcoholic liver disease is of definite predictive value for the development of cirrhosis. 相似文献
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