Initiation of DNA Synthesis and Progression through the S Period

Cell Growth ◽  
1982 ◽  
pp. 355-364 ◽  
Author(s):  
David M. Prescott
Keyword(s):  
Dna Synthesis ◽  
S Period

Radiation effects on DNA synthesis in a defined chromosomal replicon

1994 ◽  
Vol 14 (3) ◽  
pp. 1901-1908
Author(s):  
J M Larner ◽  
H Lee ◽  
J L Hamlin
Keyword(s):  
Dna Synthesis ◽  
Cell Line ◽  
Radiation Effects ◽  
Signal Transduction Pathway ◽  
Mapping Method ◽  
G1 Arrest ◽  
Tumor Suppressor P53 ◽  
Damage Sensing ◽  
Direct Demonstration ◽  
S Period

It has recently been shown that the tumor suppressor p53 mediates a signal transduction pathway that responds to DNA damage by arresting cells in the late G1 period of the cell cycle. However, the operation of this pathway alone cannot explain the 50% reduction in the rate of DNA synthesis that occurs within 30 min of irradiation of an asynchronous cell population. We are using the amplified dihydrofolate reductase (DHFR) domain in the methotrexate-resistant CHO cell line, CHOC 400, as a model replicon in which to study this acute radiation effect. We first show that the CHOC 400 cell line retains the classical acute-phase response but does not display the late G1 arrest that characterizes the p53-mediated checkpoint. Using a two-dimensional gel replicon-mapping method, we then show that when asynchronous cultures are irradiated with 900 cGy, initiation in the DHFR locus is completely inhibited within 30 min and does not resume for 3 to 4 h. Since initiation in this locus occurs throughout the first 2 h of the S period, this result implies the existence of a p53-independent S-phase damage-sensing pathway that functions at the level of individual origins. Results obtained with the replication inhibitor mimosine define a position near the G1/S boundary beyond which cells are unable to prevent initiation at early-firing origins in response to irradiation. This is the first direct demonstration at a defined chromosomal origin that radiation quantitatively down-regulates initiation.

scholarly journals CHANGES IN THE DNA SYNTHESIS PATTERN OF PARAMECIUM WITH INCREASED CLONAL AGE AND INTERFISSION TIME

1974 ◽  
Vol 61 (3) ◽  
pp. 591-598 ◽  
Author(s):  
Joan Smith-Sonneborn ◽  
Michael Klass
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Unicellular Organism ◽  
Time Intervals ◽  
Synchronized Cells ◽  
Different Ages ◽  
Self Fertilization ◽  
Autoradiographic Analysis ◽  
S Period ◽  
First Time

The clonal age in paramecia refers to the total number of vegetative divisions a clone has undergone since its origin at autogamy (self-fertilization). As clonal age increases, the interfission time usually increases. The DNA synthesis pattern of cells of different ages was compared by autoradiographic analysis of the DNA synthesis of synchronized cells at various time intervals during the cell cycle (from one division to the next). The study showed that the G1 period (the lag in DNA synthesis post division) was constant, irrespective of interfission time or clonal age; but the duration of the DNA synthesis period increased with increased interfission time or clonal age. Therefore, we have shown for the first time that the G1 period is fixed, and the S period is increased in a eukaryotic unicellular organism as a function of interfission time and clonal age.

Division and DNA Synthesis in the Kinetoplast of Crithidia Fasciculata

1969 ◽  
Vol 4 (3) ◽  
pp. 611-620
Author(s):  
W. ANDERSON ◽  
G. C. HILL
Keyword(s):  
Dna Synthesis ◽  
Crithidia Fasciculata ◽  
S Period

In Crithidia fasciculata division of the kinetoplast occurs prior to karyokinesis. Kinetoplast division involves the lateral replication of the nucleoid, and the subsequent constriction and ‘pinching off’ of the nucleoid and mitochondrial components. Two subunits are produced. DNA synthesis in the kinetoplast and the nucleus is overlapping but asynchronous. Experiments for short labelling times indicate that the kinetoplast S period either precedes, succeeds or is simultaneous with the nuclear period of DNA synthesis. The kinetoplast divides in a manner in which some radioactivity is transmitted to each unit.

Cytofluorimetric analysis of nuclear DNA during meiosis, fertilization and macronuclear development in the ciliate Tetrahymena pyriformis, syngen 1

1975 ◽  
Vol 17 (3) ◽  
pp. 471-493 ◽  
Author(s):  
F.P. Doerder ◽  
L.E. Debault
Keyword(s):  
Cell Division ◽  
Dna Synthesis ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Tetrahymena Pyriformis ◽  
Cell Cycles ◽  
Macronuclear Development ◽  
Sister Chromatids ◽  
Nuclear Development ◽  
S Period

Fluorescence cytophotometry was used to study nuclear DNA content and synthesis patterns during meiosis, fertilization and macronuclear development in the ciliated protozoon, Tetrahymena pyriformis, syngen 1. It was found that cells entered conjugation with a G1 (45C) macronucleus and a G2 (4C) micronucleus. During meiosis the micronucleus was reduced to 4 haploid nuclei, each with a 1C amount of DNA; each meiotic product then replicated to 2C, but only the nucleus next to the attachment membrane in each conjugant divided to form the two 1C gametic nuclei. The gametic nuclei replicated to 2C prior to fertilization; hence there was no S-period in the 4C fertilization nucleus (synkaryon). The first postzygotic division products immediately entered an S-period to become 4C, and at the second postzygotic division, each of the two 4C nuclei in each conjugant divided to form one 2C micronucleus and one 2C macronuclear Anlage. The macronuclear Anlagen began DNA synthesis immediately and were about 8C at the completion of conjugation; the micronuclei did not undergo rapid DNA doubling and measured between 2C and 3C when the conjugants separated. The old macronucleus did not participate in any S-period during conjugation and began to decompose after the second postzygotic division; it contained an average of 24C at the end of conjugation. From this sequence of nuclear divisions a pattern emerges that, unless a general cytoplasmic signal for DNA synthesis is suppressed, DNA synthesis always occurs in micronuclear division products immediately following separation of sister chromatids. Nuclear development continued in the first two cell cycles after conjugation. In exconjugants (the first cycle), macronuclear Anlagen underwent two rounds of DNA synthesis to become 32C and both micronuclei also underwent DNA synthesis. However, prior to the first cell division, one micronucleus and the old macronucleus completely disintegrated, and at the first cell division the remaining 4C micronucleus divided and one macronuclear Anlage was distributed to each resulting caryonide. At the end of the second cell cycle, the dividing macronucleus of each caryonide contained about 128C. These results relate to the question of ploidy of macronuclear subunits. It is argued that the G1 macronucleus contains 22 or 23 diploid subunits, each subunit being a copy of the diploid micronuclear genome. It is suggested that unequal macronuclear division relates to the question of subunit ploidy by playing a role in the phenomenon of macronuclear assortment.

scholarly journals THE RELATIONSHIP BETWEEN DEOXYRIBONUCLEIC ACID REPLICATION AND CELL DIVISION IN HEAT-SYNCHRONIZED TETRAHYMENA

1970 ◽  
Vol 46 (3) ◽  
pp. 533-543 ◽  
Author(s):  
William R. Jeffery ◽  
Kenneth D. Stuart ◽  
Joseph Frankel
Keyword(s):  
Cell Division ◽  
Heat Shock ◽  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Heat Shock Treatment ◽  
Heat Shocks ◽  
Prolonged Heat ◽  
S Period ◽  
C 50 ◽  
The Relationship

The effect of supraoptimal temperature on macronuclear DNA synthesis in Tetrahymena was studied by radioautography during prolonged heat and heat-shock synchronization treatments. Prolonged heat treatments (34°C) delayed the initiation of S, but did not appreciably delay DNA synthesis in progress. Return to optimal temperature (28°C) 50 or 100 min later resulted in initiation of S, in delayed cells, at a rate greater than in controls. During the synchronization treatment, most cells were unable to enter S during a heat shock, but initiated S with a slight delay during the following intershock period. These cells were not appreciably delayed in completion of S by subsequent heat shocks. Supraoptimal temperature appears to affect the DNA synthetic cycle near the G1 to S transition. Cells subjected to the heat-shock treatment in early G1 all participated in one S period, and many underwent a succession of two S periods. DNA synthesis occurred in about 50% of the cells between EST and the first synchronous division, with the likelihood of DNA synthesis becoming greater the longer the interval between these two events. In some cells no detectable DNA synthesis occurred between EST and the second synchronous division. It was concluded that a precise temporal alternation of DNA replication and cell division is not obligatory in Tetrahymena.

scholarly journals Cell cycle-dependent expression of thymidylate synthase in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (12) ◽  
pp. 2858-2864 ◽  
Author(s):  
R K Storms ◽  
R W Ord ◽  
M T Greenwood ◽  
B Mirdamadi ◽  
F K Chu ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Dna Synthesis ◽  
Thymidylate Synthase ◽  
S Phase ◽  
Mrna Levels ◽  
Activity Profile ◽  
S Period ◽  
Cycle Dependent ◽  
Specific Mrna

Synchronous populations of Saccharomyces cerevisiae cells, generated by two independent methods, have been used to show that thymidylate synthase, in contrast to the vast majority of cellular proteins thus far examined, fluctuates periodically during the S. cerevisiae cell cycle. The enzyme, as assayed by two different methods, accumulated during S period and peaked in mid to late S phase, and then its level dropped. These observations suggest that both periodic synthesis and the instability of the enzyme contribute to the activity profile seen during the cell cycle. Accumulation of thymidylate synthase is determined at the level of its transcript, with synthase-specific mRNA levels increasing at least 10-fold to peak near the beginning of S period and then falling dramatically to basal levels after the onset of DNA synthesis. This mRNA peak coincided with the time during the cell cycle when thymidylate synthase levels were increasing maximally and immediately preceded the peak of DNA synthesis, for which the enzyme provides precursor dTMP.

scholarly journals RADIOAUTOGRAPHIC STUDIES OF PLAQUE-FORMING CELLS

1968 ◽  
Vol 128 (2) ◽  
pp. 235-257 ◽  
Author(s):  
Aurelia M. C. Koros ◽  
John M. Mazur ◽  
Margaret J. Mowery
Keyword(s):  
Cell Proliferation ◽  
Dna Synthesis ◽  
Tritiated Thymidine ◽  
Red Cells ◽  
G1 Phase ◽  
Spleen Cells ◽  
Antigen Dose ◽  
Sheep Red Cells ◽  
Large Populations ◽  
S Period

A method has been described for obtaining radioautographs of plaque-forming cells. The method permits radioautographic analyses of small numbers of plaque-forming cells amidst large populations of non-plaque-forming cells. Spleen cells that were pulse-labeled with tritiated thymidine could be categorized readily as labeled or not labeled. Using this method it was found that (a) at least 55% of plaque-forming cells which appear 3 days after a maximal stimulus of 4 x 108 sheep red cells are still capable of DNA synthesis, and must have arisen by cell proliferation; (b) the rate of proliferation of plaque-forming cells is proportional to the log of the dose of antigen; (c) the S period of plaque-forming cells is at least 2 hr, appears to be constant, and is not influenced by antigen dose. The results suggest that antigen stimulates proliferation of plaque-forming cells by hastening their transit through the G1 phase of the generative cycle.

Comparative Analysis of the Kinetics of DNA Synthesis after Exposure during Different Phases of the Cell Cycle S Period

2013 ◽  
Vol 156 (2) ◽  
pp. 260-265 ◽  
Author(s):  
I. P. Shabalkin ◽  
E. Yu. Grigor’eva ◽  
P. I. Shabalkin ◽  
M. V. Gudkova ◽  
A. S. Yagubov
Keyword(s):  
Cell Cycle ◽  
Comparative Analysis ◽  
Dna Synthesis ◽  
S Period ◽  
Kinetics Of

scholarly journals ANALYSIS OF THE SCHEDULE OF DNA REPLICATION IN HEAT-SYNCHRONIZED TETRAHYMENA

1973 ◽  
Vol 59 (1) ◽  
pp. 1-11 ◽  
Author(s):  
William R. Jeffery ◽  
Joseph Frankel ◽  
Lawrence E. de Bault ◽  
Leslie M. Jenkins
Keyword(s):  
Cell Cycle ◽  
High Temperature ◽  
Dna Replication ◽  
Heat Shock ◽  
Dna Synthesis ◽  
Shock Treatment ◽  
Heat Shock Treatment ◽  
Dividing Cells ◽  
S Period ◽  
Selection Of

The temporal schedule of DNA replication in heat-synchronized Tetrahymena was studied by autoradiographic and cytofluorometric methods. It was shown that some cells, which were synchronized by selection of individual dividing cells or by temporary thymidine starvation, incorporated [3H]thymidine into macronuclei in a periodic fashion during the heat-shock treatment. It was concluded that supernumerary S periods occurred while cell division was blocked by high temperature. The proportion of cells which initiated supernumerary S periods was found to be dependent on the duration of the heat-shock treatment and on the cell cycle stage when the first heat shock was applied. Cytofluorometric measurements of Feulgen-stained macronuclei during the heat-shock treatment indicated that the DNA complement of these cells was substantially increased and probably duplicated during the course of each S period. Estimates of DNA content also suggested that the rate of DNA synthesis progressively declined during long heat-shock treatments. These results indicate that the mechanism which brings about heat-induced division synchrony is not an interruption of the process of DNA replication. Further experiments were concerned with the regulation of DNA synthesis during the first synchronized division cycle. It was shown that participation in DNA synthesis at this time increased as more cells were able to conclude the terminal S period during the preceding heat-shock treatment. It is suggested that a discrete period of time is necessary after the completion of DNA synthesis before another round of DNA synthesis can be initiated.

scholarly journals INHIBITION OF MYOBLAST FUSION AFTER ONE ROUND OF DNA SYNTHESIS IN 5-BROMODEOXYURIDINE

1970 ◽  
Vol 44 (1) ◽  
pp. 134-150 ◽  
Author(s):  
Richard Bischoff ◽  
Howard Holtzer
Keyword(s):  
Dna Synthesis ◽  
Cell Number ◽  
S Phase ◽  
Minor Effect ◽  
Myogenic Cells ◽  
Normal Morphology ◽  
Low Concentrations ◽  
S Period ◽  
A Minor ◽  
Multinucleated Myotubes

The thymidine analogue 5-bromodeoxyuridine (BUdR) has a differential effect on the synthesis of tissue-specific products and molecules required for growth and division. Proliferating myogenic cells cultured in BUdR fail to fuse and fail to initiate the synthesis of contractile protein filaments. Conversely, BUdR has but a minor effect on cell viability and reproductive integrity. Low concentrations of BUdR result in an enhancement of cell number relative to the controls; higher concentrations are cytotoxic. Suppression of myogenesis is reversible after at least 10 cell generations of growth in the analogue. Cells that do not synthesize DNA, such as postmitotic myoblasts and myotubes, are not affected by BUdR. Incorporation of BUdR for one round of DNA synthesis was accomplished by first incubating myogenic cells, prior to fusion, in 5-fluorodeoxyuridine (FUdR) to block DNA synthesis and collect cells in the presynthetic phase. The cells were then allowed to synthesize either normal DNA or BU-DNA for one S period by circumventing the FUdR block with BUdR or BUdR plus thymidine (TdR). The cultures were continued in FUdR to prevent dilution of the incorporated analogue by further division. After 3 days, the cultures from the FUdR-BUdR series showed the typical BUdR effect; the cells were excessively flattened and few multinucleated myotubes formed. Cells in the control cultures were of normal morphology, and multinucleated myotubes were present. These results were confirmed in another experiment in which BUdR-3H was added to 2-day cultures in which myotubes were forming. Fusion of thymidine-3H-labeled cells begins at 8 hr after the preceding S phase. In contrast, cells which incorporate BUdR-3H for one S period do not fuse with normal myotubes.

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