Nuclear Morphology and the Ultra-Structural Localization of Deoxyribonucleic Acid Synthesis during Interphase

1969 ◽  
Vol 4 (3) ◽  
pp. 569-582
Author(s):  
GILLIAN R. MILNER
Keyword(s):  
Epithelial Cells ◽  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Ultrastructural Localization ◽  
Cell Types ◽  
Acid Synthesis ◽  
Lung Fibroblasts ◽  
Nuclear Morphology ◽  
Structural Localization ◽  
Electron Microscope Autoradiography

The ultrastructural localization of deoxyribonucleic acid (DNA) synthesis was studied by electron-microscope autoradiography in human transforming lymphocytes, embryonic lung fibroblasts, epithelial cells and normoblasts. Euchromatin was found to be active in DNA synthesis in all cell types studied, whereas heterochromatin was inactive. However, DNA synthesis was also prominent in the regions where heterochromatin was thought to be decondensing to form euchromatin. Analysis of sequential changes in nuclear morphology of the transforming lymphocyte suggested that there is decondensation of heterochromatin during the S-phase until none is left. In nuclei with no heterochromatin a prominent localization of DNA synthesis was at the nuclear membrane. This sequence of complete decondensation of heterochromatin also seemed likely for fibroblasts and epithelial cells. Normoblasts however showed no stage where the nucleus was wholly euchromatic and it is suggested that in this cell decondensation of heterochromatin for replication is localized and transient.

scholarly journals Correlation between nuclear morphology and rate of deoxyribonucleic acid synthesis in a normal cell line.

1979 ◽  
Vol 27 (1) ◽  
pp. 188-192 ◽  
Author(s):  
P Dörmer ◽  
W Abmayr
Keyword(s):  
Dna Synthesis ◽  
Cell Line ◽  
Chinese Hamster ◽  
Cell Types ◽  
Acid Synthesis ◽  
High Rate ◽  
Second Step ◽  
Synthesis Rate ◽  
Nuclear Morphology ◽  
Deoxyribonucleic Acid Synthesis

Chinese hamster fibroblasts were investigated for the existence of correlations between proliferative activity and nuclear morphology. As a proliferative parameter, the rate of DNA synthesis of individual cells was determined by quantitative 14C-autoradiography. In a second step the images of the Feulgen-stained nuclei were digitized for extraction of features of morphology and texture. These features were correlated with the corresponding DNA synthesis rate values. The following relationships were found: Round nuclei have higher rates of DNA synthesis than flat ones. The more chromatin is packed at the nuclear rim, possibly representing heterochromatin, the lower the rate of DNA synthesis. The DNA synthesis rate also correlates with the graininess of chromatin. Larger areas of condensed chromatin are associated with lower rate values. A fine and irregular network of chromatin, as is typical of immature cell types, is associated with a high rate of DNA synthesis. Although these results are presently confined to the cell line investigated, parallels seem to exist to other cell types, such as erythropoietic cells, which await further investigation.

scholarly journals Ornithine decarboxylase activity and the onset of deoxyribonucleic acid synthesis in regenerating liver

1978 ◽  
Vol 170 (1) ◽  
pp. 123-127 ◽  
Author(s):  
J A McGowan ◽  
N Fausto
Keyword(s):  
Dna Synthesis ◽  
Ornithine Decarboxylase ◽  
Time Course ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
Decarboxylase Activity ◽  
Ornithine Decarboxylase Activity ◽  
Low Protein ◽  
Deoxyribonucleic Acid Synthesis ◽  
Second Peak

Compared with normally fed animals, rats fed on a low-protein diet for 3 days exhibit a considerable delay in DNA synthesis after partial hepatectomy. In the regenerating livers of these animals (a) the timing of the first peak of ornithine decarboxylase activity is not altered and (b) the second peak of enzyme activity is delayed by a few hours, but polyamine concentrations are similar to those of normally fed rats. The results suggest that regardless of the possible effect of polyamines on DNA synthesis, the time course of ornithine decarboxylase activity appears to be independent of the onset of DNA replication in regenerating livers.

scholarly journals Effect of drugs on deoxyribonucleic acid synthesis in isolated mammalian cell nuclei. Comparison with partially purified deoxyribonucleic acid polymerases

1979 ◽  
Vol 178 (3) ◽  
pp. 621-626 ◽  
Author(s):  
J F Burke ◽  
P M Duff ◽  
C K Pearson
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
Cell Nuclei ◽  
Isolated Nuclei ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta ◽  
Deoxyribonucleic Acid Synthesis

In order to ascertain the identity of the DNA-dependent DNA polymerase responsible for the observed DNA synthesis in nuclei isolated from baby-hamster kidney (BHK-21/C13) cells a comparative study was carried out on the effects of some drugs, reported to influence DNA synthesis, on DNA synthesis catalysed by these nuclei and by partially purified DNA polymerase-alpha and -beta. In all cases DNA synthesis by isolated nuclei and polymerase-alpha was inhibited to similar extents by N-ethylmaleimide, p-hydroxymercuribenzoate, novobiocin, heparin and phosphonoacetic acid; polymerase-beta was much less affected by these compounds. Ethidium bromide inhibited all DNA synthesis to similar extents, although at low concentrations (about 2 microgram/ml) synthesis in isolated nuclei was stimulated. The results are discussed in relation to the proposal that DNA polymerase-alpha catalyses the covalent extension of Okazaki fragments that these nuclei carry out in vitro.

Discordant regulatory changes in monocrotaline-induced megalocytosis of lung arterial endothelial and alveolar epithelial cells

2006 ◽  
Vol 290 (6) ◽  
pp. L1216-L1226 ◽  
Author(s):  
Somshuvra Mukhopadhyay ◽  
Pravin B. Sehgal
Keyword(s):  
Cell Cycle ◽  
Epithelial Cells ◽  
Dna Synthesis ◽  
A549 Cells ◽  
Cell Types ◽  
Alveolar Epithelial Cells ◽  
Cell Type ◽  
Alveolar Epithelial ◽  
Unfolded Protein ◽  
Cell Cycle Regulatory Proteins

Monocrotaline (MCT) causes pulmonary hypertension in the rat by a mechanism characterized by megalocytosis (enlarged cells with enlarged endoplasmic reticulum and Golgi and a cell cycle arrest) of pulmonary arterial endothelial (PAEC), arterial smooth muscle, and type II alveolar epithelial cells. In cell culture, although megalocytosis is associated with a block in entry into mitosis in both lung endothelial and epithelial cells, DNA synthesis is stimulated in endothelial but inhibited in epithelial cells. The molecular mechanism(s) for this dichotomy are unclear. While MCTP-treated PAEC and lung epithelial (A549) cells both showed an increase in the “promitogenic” transcription factor STAT3 levels and in the IL-6-induced nuclear pool of PY-STAT3, this was transcriptionally inactive in A549 but not in PAEC cells. This lack of transcriptional activity of STAT3 in A549 cells correlated with the cytoplasmic sequestration of the STAT3 coactivators CBP/p300 and SRC1/NcoA in A549 cells but not in PAEC. Both cell types displayed a Golgi trafficking block, loss of caveolin-1 rafts, and increased nuclear Ire1α, but an incomplete unfolded protein response (UPR) with little change in levels of UPR-induced chaperones including GRP78/BiP. There were discordant alterations in cell cycle regulatory proteins in the two cell types such as increase in levels of both cyclin D1 and p21 simultaneously, but with a decrease in cdc2/cdk1, a kinase required for entry into mitosis. While both cell types showed increased cytoplasmic geminin, the DNA synthesis-initiating protein Cdt1 was predominantly nuclear in PAEC but remained cytoplasmic in A549 cells, consistent with the stimulation of DNA synthesis in the former but an inhibition in the latter cell type. Thus differences in cell type-specific alterations in subcellular trafficking of critical regulatory molecules (such as CBP/p300, SRC1/NcoA, Cdt1) likely account for the dichotomy of the effects of MCTP on DNA synthesis in endothelial and epithelial cells.

scholarly journals Deoxyribonucleic acid synthesis in mammalian nuclei. Incorporation of deoxyribonucleotides and chain-terminating nucleotide analogues

1971 ◽  
Vol 121 (5) ◽  
pp. 803-809 ◽  
Author(s):  
M. A. Waqar ◽  
L. A. Burgoyne ◽  
M. R. Atkinson
Keyword(s):  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
Deoxyribonucleic Acid Synthesis ◽  
Dna Chain ◽  
Relationship Of ◽  
The Relationship ◽  
Neighbour Analysis

The properties of a nuclear preparation from rat liver and thymus are described. (1) Nearest-neighbour analysis after incorporation of 32P-labelled nucleotide residues from dATP, dCTP, dGTP, dTTP and arabinofuranosyl analogues of CTP and ATP shows template-dependent DNA synthesis. (2) Where primer termini are limiting, incorporation of arabinofuranosyl analogues of AMP and CMP residues proceeds to a limit indicating that both of these analogues are DNA chain terminators. (3) No large differences have been found between the priming potentialities or the intrinsic DNA polymerase activities of nuclei from resting or regenerating liver and the relationship of this DNA synthesis in vitro to DNA replication or repair in vivo is briefly discussed.

USE OF I125-LABELED 5-IODO-2′-DEOXYURIDINE FOR THE MEASUREMENT OF DNA SYNTHESIS IN MAMMALIAN CELLS IN VITRO

1963 ◽  
Vol 41 (11) ◽  
pp. 2343-2351 ◽  
Author(s):  
S. Mak ◽  
J. E. Till
Keyword(s):  
Dna Synthesis ◽  
Mammalian Cells ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
X Rays ◽  
Beta Particles ◽  
Deoxyribonucleic Acid Synthesis ◽  
Gamma Emitter

The use of isotopically labeled 5-iodo-2′-deoxyuridine (I125UdR) for determination of the rate of deoxyribonucleic acid synthesis in mammalian cells in vitro has been investigated. The results obtained indicate that for this purpose I125UdR is a suitable substitute for the more commonly used DNA precursor, tritium-labeled thymidine (H3TdR). I125UdR appears to be incorporated specifically into the DNA of cells in culture, and has been demonstrated to compete with H3TdR, although the Km for H3TdR was smaller than that of I125UdR by a factor of approximately 4. The amount of label incorporated into DNA of cells increased linearly with time. When the rate of DNA synthesis was reduced by exposure of the cells to various doses of X-rays, the ratio of I125UdR incorporation to H3TdR incorporation into DNA of cells was found to be a constant, which supports the view that uptake of the analogue provides as reliable an indication of effects upon the rate of DNA synthesis as does that of H3TdR. The chief advantage of I125UdR over H3TdR is that I125 is a gamma emitter, so that the difficulties encountered in detection of the low energy beta particles from H3 may be avoided.

scholarly journals TRANSIENT PHOSPHORYLATION OF DEOXYRIBOSIDES AND REGULATION OF DEOXYRIBONUCLEIC ACID SYNTHESIS

1961 ◽  
Vol 11 (2) ◽  
pp. 311-319 ◽  
Author(s):  
Yasuo Hotta ◽  
Herbert Stern
Keyword(s):  
Dna Synthesis ◽  
Enzyme Induction ◽  
Deoxyribonucleic Acid ◽  
Lilium Longiflorum ◽  
Acid Synthesis ◽  
Deoxyribonucleic Acid Synthesis ◽  
Patterns Of Behavior

Microspores isolated from Lilium longiflorum and Trillium erectum were studied with respect to their capacities for phosphorylating deoxyribosides in vitro. It was found that such capacities are manifest only during brief intervals of time adjacent to periods of DNA synthesis, and that none of the neighboring cells in the anther acquire them. The observed patterns of behavior are interpreted in terms of enzyme induction as a device for regulating DNA synthesis.

Rat mesothelial and tracheal epithelial cells show equal DNA sensitivity to hydrogen peroxide-induced oxidant injury

1995 ◽  
Vol 268 (5) ◽  
pp. L832-L838 ◽  
Author(s):  
A. Churg ◽  
B. Keeling ◽  
B. Gilks ◽  
S. Porter ◽  
P. Olive
Keyword(s):  
Hydrogen Peroxide ◽  
Epithelial Cells ◽  
Similar Pattern ◽  
Mesothelial Cell ◽  
Cell Types ◽  
Lung Fibroblasts ◽  
Continuous Exposure ◽  
Strand Breaks ◽  
Oxidant Damage ◽  
Tracheal Epithelial

To study the relative sensitivity of rat tracheal epithelial and mesothelial cell DNA to oxidant damage, we used the comet assay, a gel microelectrophoresis method that allows visual determination of DNA strand breaks on a cell-by-cell basis, to evaluate damage after hydrogen peroxide exposure. By both a qualitative and a quantitative assay, tracheal epithelial mesothelial cells demonstrated a similar dose-response increase in the number of cells showing strand breaks and the number of breaks per cell after exposure to increasing concentrations of hydrogen peroxide; but even at the highest concentration, some cells failed to show damage. By contrast, 100% of cultured V79 lung fibroblasts showed evidence of damage. Catalase and deferoxamine largely prevented the formation of strand breaks, while superoxide dismutase was not protective. To evaluate DNA repair, cells were exposed to 10 microM hydrogen peroxide for 10 min, washed, and maintained in culture medium; by 2 h the proportion of mesothelial and epithelial cells showing comets had returned to control levels for both cell types. Both cell types also showed a similar pattern of increasing damage after continuous exposure to 10 microM hydrogen peroxide for periods up to 2 h. We conclude that, in this system, 1) mesothelial and tracheobronchial epithelial cells show a similar pattern of DNA injury and repair after hydrogen peroxide exposure; 2) hydrogen peroxide damages DNA of both cell types via a mechanism probably related to the iron-catalyzed formation of hydroxyl radical; and 3) both types of cells appear to be heterogeneous in their sensitivity to oxidant damage, with some cells showing extreme resistance to such damage.

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