Contribution of DNA polymerase δ to DNA replication in permeable CHO cells synchronized in S phase

Basnakian, Alexei G.; Bánfalvi, Gáspár; Sarkar, Nirmal Kumar

doi:10.1093/nar/17.12.4757

Cited by 37 publications

(15 citation statements)

References 31 publications

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“…The cells in each fraction were divided into 17 subfractions to cover the total area of the cytometric profile. C-values were calculated from the appropriate area under the flow cytometric profile and were averaged to yield the DNA content for each fraction as described previously [18].…”

Section: Facs Analysis Of Cell Cyclementioning

confidence: 99%

Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B-cells

et al. 2007

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Murine pre-B-cells grown in the presence of lower (1 microM) or higher (5 microM) concentration of cadmium chloride were separated into 13 fractions by centrifugal elutriation. The rate of DNA synthesis after cadmium treatment determined in permeable cells was dependent on cell culture density during cadmium treatment. Cell cycle analysis revealed a shift in the profile of DNA synthesis from replicative to repair DNA synthesis upon cadmium treatment. The study of the relationship between cell culture density and cell diameter at lower and higher cell densities in the presence of 1 microM cadmium chloride concentration showed that a. at 5 x 10(5) cell/ml or lower densities cells were shrinking indicating apoptotic changes, b. at higher cell culture densities the average cell size increased, c. the treatment of cells with low CdCl(2) concentration (1 microM) at higher cell culture density (>5 x 10(5) cell/ml) did not change significantly the average cell diameter. At 5 microM cadmium concentration and higher cell culture densities (>5 x 10(5) cell/ml) the average cell size decreased in each elutriated fraction. Most significant inhibition of cell growth took place in early S phase (2.0-2.5 C value). Apoptotic chromatin changes in chromatin structure after cadmium treatment were seen as large extensive disruptions, holes in the nuclear membrane and stickiness of incompletely folded chromosomes.

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Section: Facs Analysis Of Cell Cyclementioning

confidence: 99%

Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B-cells

et al. 2007

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“…C-values were calculated from the appropriate area under the flow cytometric profile and were averaged to yield the DNA content for each fraction as described previously. 10 …”

Section: Facs Analysis Of Cell Cyclementioning

confidence: 99%

Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

et al. 2005

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CHO cells were grown in the presence of 1 mu M CdCl(2) and subjected to ATP-dependent replicative DNA synthesis after permeabilization. By decreasing the density of the cell culture replicative DNA synthesis was diminishing. At higher than 2 x 10(6) cell/ml concentration Cd had virtually no effect on the rate of DNA replication. Growth at higher cell concentrations could be suppressed by increasing Cd concentration. After Cd treatment cells were synchronized by counterflow centrifugal elutriation. Cadmium toxicity on cell growth in early and mid S phase led to the accumulation of enlarged cells in late S phase. Flow cytometry showed increased cellular and nuclear sizes after Cd treatment. As the cells progressed through the S phase, 11 subpopulations of nuclear sizes were distinguished. Apoptotic chromatin changes were visualized by fluorescent microscopy in a cell cycle dependent manner. In the control untreated cells the main transitory forms of chromatin corresponded to those we have published earlier (veil-like, supercoiled chromatin, fibrous, ribboned structures, chromatin strings, elongated prechromosomes, precondensed chromosomes). Cadmium treatment caused: (a) the absence of decondensed veil-like structures and premature chromatin condensation in the form of apoptotic bodies in early S phase (2.2-2.4 average C-value), (b) the absence of fibrous structures, the lack of supercoiled chromatin, the appearance of uncoiled ribboned chromatin and perichromatin semicircles, in early mid S phase (2.5-2.9 C), (c) the presence of perichromatin fibrils and chromatin bodies in mid S phase (2.9-3.2 C), (d) early intra-nuclear inclusions, elongated forms of premature chromosomes, the extrusion and rupture of nuclear membrane later in mid S phase (3.3-3.4 C), (e) the exclusion of chromatin bodies and the formation of clusters of large-sized perichromatin granules in late S phase (3.5-3.8 C) and (f) large extensive disruptions and holes in the nuclear membrane and the clumping of incompletely folded chromosomes (3.8-4. C).

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“…The fraction collected during loading, which contained dead cells, was discarded. The last fractions (fractions [19][20] could not be used either due to their heterogeneity and low cell number. Each fraction was routinely monitored by light microscopy and even fractions were analyzed by fluorescence activated cell sorting (FACS).…”

Section: Elutriation Of Cellular Fractionsmentioning

confidence: 99%

Section: Facs Analysis Of Cell Cyclementioning

confidence: 99%

Preapoptotic chromatin changes induced by ultraviolet B irradiation in human erythroleukemia K562 cells

et al. 2007

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Exponentially growing human erythroleukemia K562 cells were permeabilized and the dose dependent decrease of DNA synthesis rate was measured after ultraviolet (UV B, 290 nm) irradiation. Cells were able to overcome 2 and 5 J/m2 UV doses, partial recovery was observed at 15 J/m2, while at high (25 J/m2) UV dose replicative DNA synthesis remained suppressed. K562 cells were subjected to synchronization prior to and after UV irradiation (24 J/m2) and 18 fractions were collected by centrifugal elutriation. Cell cycle analysis by flow cytometry did not show early apoptotic cells after UV irradiation. The gradual increase in DNA content typical for non-irradiated cells was contrasted by an early S phase block between 2.2 and 2.4 C-values after UV irradiation. Cell cycle dependent chromatin changes after ultraviolet irradiation were seen as a fine fibrillary network covering the mainly fibrous chromatin structures and incompletely folded primitive chromosomes. Based on observations after UV irradiation and on earlier results with cadmium treatment and gamma irradiation, we confirm that typical chromatin changes characteristic to genotoxic agents can be recognized and classified.

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Contribution of DNA polymerase δ to DNA replication in permeable CHO cells synchronized in S phase

Cited by 37 publications

References 31 publications

Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B-cells

Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B-cells

Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

Preapoptotic chromatin changes induced by ultraviolet B irradiation in human erythroleukemia K562 cells

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