scholarly journals Inhibitor effects during the cell cycle in Chlamydomonas reinhardtii. Determination of transition points in asynchronous cultures.

1975 ◽  
Vol 67 (1) ◽  
pp. 126-135 ◽  
Author(s):  
S H Howell ◽  
W J Blaschko ◽  
C M Drew
Keyword(s):  
Cell Cycle ◽  
Protein Synthesis ◽  
Cell Division ◽  
Chlamydomonas Reinhardtii ◽  
Macromolecular Synthesis ◽  
Synchronous Cultures ◽  
Organellar Dna ◽  
Rna And Protein Synthesis ◽  
Transition Points

A wide variety of inhibitors (drugs, antibiotics, and antimetabolites) will block cell division within an ongoing cell cycle in autotrophic cultures of Chlamydomonas reinhardtii. To determine when during the cell cycle a given inhibitor is effective in preventing cell division, a technique is described which does not rely on the use of synchronous cultures. The technique permits the measurement of transition points, the cell cycle stage at which the subsequent cell division becomes insensitive to the effects of an inhibitor. A map of transition points in the cell cycle reveals that they are grouped into two broad periods, the second and fourth quarters. In general, inhibitors which block organellar DNA, RNA, and protein synthesis have second-quarter transition points, while those which inhibit nuclear cytoplasmic macromolecular synthesis have fourth-quarter transition points. The specific grouping of these transition points into two periods suggests that the synthesis of organellar components is completed midway through the cell cycle and that the synthesis of nonorganellar components required for cell division is not completed until late in the cell cycle.

scholarly journals To Divide or Not to Divide? How Deuterium Affects Growth and Division of Chlamydomonas reinhardtii

Biomolecules ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 861
Author(s):  
Veronika Kselíková ◽  
Vilém Zachleder ◽  
Kateřina Bišová
Keyword(s):  
Cell Cycle ◽  
Chlamydomonas Reinhardtii ◽  
Cell Cycle Progression ◽  
Model Organism ◽  
Cycle Progression ◽  
Synchronous Cultures ◽  
Multiple Fission ◽  
First Choice ◽  
High Doses

Extensive in vivo replacement of hydrogen by deuterium, a stable isotope of hydrogen, induces a distinct stress response, reduces cell growth and impairs cell division in various organisms. Microalgae, including Chlamydomonas reinhardtii, a well-established model organism in cell cycle studies, are no exception. Chlamydomonas reinhardtii, a green unicellular alga of the Chlorophyceae class, divides by multiple fission, grows autotrophically and can be synchronized by alternating light/dark regimes; this makes it a model of first choice to discriminate the effect of deuterium on growth and/or division. Here, we investigate the effects of high doses of deuterium on cell cycle progression in C. reinhardtii. Synchronous cultures of C. reinhardtii were cultivated in growth medium containing 70 or 90% D2O. We characterize specific deuterium-induced shifts in attainment of commitment points during growth and/or division of C. reinhardtii, contradicting the role of the “sizer” in regulating the cell cycle. Consequently, impaired cell cycle progression in deuterated cultures causes (over)accumulation of starch and lipids, suggesting a promising potential for microalgae to produce deuterated organic compounds.

Oxygen uptake during the cell cycle of the fission yeast Schizosaccharomyces pombe

1978 ◽  
Vol 33 (1) ◽  
pp. 399-411
Author(s):  
J. Creanor
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Oxygen Uptake ◽  
Dna Synthesis ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nuclear Division ◽  
Synchronous Culture ◽  
Synchronous Cultures ◽  
The Many

Oxygen uptake was measured in synchronous cultures of the fission yeast Schizosaccharomyces pombe. The rate of oxygen uptake was found to increase in a step-wise manner at the beginning of the cycle and again in the middle of the cycle. The increases in rate were such that overall, oxygen uptake doubled in rate once per cell cycle. Addition of inhibitors of DNA synthesis or nuclear division to a synchronous culture did not affect the uptake of oxygen. In an induced synchronous culture, in which DNA synthesis, cell division, and nuclear division, but not ‘growth’ were synchronized, oxygen uptake increased continuously in rate and did not show the step-wise rises which were shown in the selection-synchronized culture. These results were compared with previous measurements of oxygen uptake in yeast and an explanation is suggested for the many different patterns which have been reported.

Wachstumsparameter in normalen und durch Actinomycin verlängerten Zellzyklen von Tetrahymena / Growth Parameters in Normal and Actinomycin-induced prolonged Cell Cycles of Tetrahymena

1969 ◽  
Vol 24 (12) ◽  
pp. 1624-1629 ◽  
Author(s):  
Günter Cleffmann
Keyword(s):  
Cell Cycle ◽  
Protein Synthesis ◽  
Cell Division ◽  
Dna Replication ◽  
Cell Growth ◽  
Rna Synthesis ◽  
Growth Parameters ◽  
Average Duration ◽  
Cell Cycles ◽  
Growing Cell

Actinomycin in low concentration (0,2 μg/ml — 0,5 μg/ml) prolongs the average duration of the cell cycle of Tetrahymena considerably, but does not inhibit cell division completely. Some parameters of the growing cell have been tested in cell cycles extended in this way and compared to those of normally growing cells. The RNA synthesis of treated cells is reduced to such an extent that the RNA content per cell decreases during the prolonged cell cycle. Nevertheless cell growth, protein synthesis and DNA replication proceed at almost the same rate as in untreated cells. These findings indicate that the presence of actinomycin does not interfere with RNA fractions necessary for growth but reduce the synthesis of RNA fractions which are essential for cell division. Therefore a longer period is needed for their accumulation.

scholarly journals GROWTH INHIBITION OF MURINE TUMOR CELLS, IN VITRO, BY PUROMYCIN, [6N]O2'-DIBUTYRYL 3',5'-ADENOSINE MONOPHOSPHATE, OR ADENOSINE

1973 ◽  
Vol 57 (2) ◽  
pp. 397-405 ◽  
Author(s):  
D. B. Thomas ◽  
Gay Medley ◽  
C. A. Lingwood
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Cell Growth ◽  
Growth Inhibition ◽  
Tumor Cells ◽  
Growth Arrest ◽  
Adenosine Monophosphate ◽  
Synchronous Cultures ◽  
Murine Tumor Cells

The cytostatic effects of puromycm, [6N]O2'-dibutyryl 3',5'-adenosine monophosphate, and adenosine on asynchronous and synchronous cultures of the murine mastocytoma, P815Y, have been studied. Cell growth was arrested after a minimum of one further division. A model is proposed for the inhibition of cell division in which the periods of inhibition and growth arrest are separated in time by one cell cycle.

Regulation of K+ Influx in Barley Roots: Evidence for Direct Control by Internal K+

1977 ◽  
Vol 4 (2) ◽  
pp. 313 ◽  
Author(s):  
ADM Glass
Keyword(s):  
Protein Synthesis ◽  
Cell Division ◽  
Actinomycin D ◽  
Direct Control ◽  
Pretreatment Period ◽  
Rna And Protein Synthesis ◽  
Lag Period ◽  
Limited Protein ◽  
K Concentration ◽  
Gamma Irradiated

Values for plasmalemma influx of K+ into excised barley roots, from solutions containing 0.05 mM KCl plus 0.5 mM CaSO4, were reduced by 50-60% following a 6-h pretreatment period in 50 mM KCl plus 0.05 mM CaSO4 solution. This reduction of influx, associated with increased internal K+ concentration, was independent of DNA, RNA and protein synthesis during the pretreatment period as indicated by its insensitivity to the presence of 5-fluorodeoxyuridine, actinomycin D, cycloheximide, p-fluorophenylalanine or anisomycin in the pretreatment solutions. Roots of plantlets grown from gamma-irradiated barley seeds, which were incapable of under-going cell division and capable of only limited protein synthesis, were nevertheless able to reduce K+ influx values in response to increased internal K+ concentration. The measurement of K+ influx from 0.05 mM KCl solutions following pretreatment periods as short as 15 min in 50 mM KCl gave no evidence of any lag period in the development of reduced influx associated with increased internal K+ status. The above experiments are discussed in terms of a model for the regulation of K+ influx which ascribes a direct 'allosteric' role to internal K+ in controlling influx.

Nucleic acid and protein synthesis and pattern regulation in hydra

Development ◽  
1969 ◽  
Vol 21 (1) ◽  
pp. 55-70
Author(s):  
S. G. Clarkson
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Experimental Approach ◽  
Direct Evidence ◽  
Animal Cells ◽  
Rna And Protein Synthesis ◽  
Metabolic Activities ◽  
Time Required ◽  
Pattern Regulation

In a previous paper (Clarkson, 1969) data were presented which indicate that hypostome determination is accompanied by a large and rapid burst of RNA synthesis, a slight stimulation of protein synthesis, and no increase in DNA synthesis. More direct evidence concerning the relative importance of these metabolic activities in hypostome determination is reported in this paper. The experimental approach made use of the transplantation test of Webster & Wolpert (1966) in conjunction with some inhibitors of DNA, RNA and protein synthesis, the rationale being that if these metabolic activities play important roles in the determination of the hypostome, then their inhibition would be expected to have severe effects on the time required for this process. Regarding the inhibitors, hydroxyurea (HU) inhibits DNA synthesis in a variety of animal cells without altering rates of formation of RNA or protein (Young & Hodas, 1964; Yarbro, Kennedy & Barnum, 1965; Schwartz, Garofalo, Sternberg & Philips, 1965).

Cell-cycle-dependent effects of sodium-n-butyrate in Physarum polycephalum

1982 ◽  
Vol 58 (1) ◽  
pp. 303-311
Author(s):  
P. Loidl ◽  
P. Grobner ◽  
A. Csordas ◽  
B. Puschendorf
Keyword(s):  
Cell Cycle ◽  
Fatty Acids ◽  
Protein Synthesis ◽  
Physarum Polycephalum ◽  
Mitotic Cycle ◽  
Short Chain Fatty Acids ◽  
Time Interval ◽  
Retarding Effect ◽  
Rna And Protein Synthesis ◽  
Cycle Dependent

Sodium-n-butyrate affects the length of the mitotic cycle of Physarum polycephalum. Application during S- or early G2-period results in a delay of the subsequent mitosis, whereas application later in the cycle has no delaying effect. Interestingly, the second mitotic cycle after application is considerably shortened when butyrate has been administered during S- or early G2-period of the preceding cycle. In comparison, other homologous short-chain fatty acids were tested; the retarding effect on mitosis increases with the number of carbon atoms, although only butyrate can shorten the second mitotic cycle. It is shown that butyrate causes an immediate depression of synthesis of DNA, RNA and protein. After a certain time-interval the plasmodium overcomes the butyrate block. DNA synthesis is fully recovered and the inhibition of RNA and protein synthesis is even overcompensated until the next mitosis, as reflected by elevated levels of RNA and protein.

Tubulin gene expression in the Chlamydomonas reinhardtii cell cycle: elimination of environmentally induced artifacts and the measurement of tubulin mRNA levels

1988 ◽  
Vol 89 (3) ◽  
pp. 397-403
Author(s):  
D.S. Nicholl ◽  
J.A. Schloss ◽  
P.C. John
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Division ◽  
Chlamydomonas Reinhardtii ◽  
Environmental Conditions ◽  
Mrna Levels ◽  
Tubulin Gene ◽  
Mrna Accumulation ◽  
Tubulin Mrna ◽  
Flagellar Proteins

To investigate the involvement of tubulin gene expression in controlling cell division events in Chlamydomonas reinhardtii we have measured tubulin mRNA levels during the cell cycle under different environmental conditions. In C. reinhardtii cells grown under the synchronizing conditions of 14 h of light followed by 10 h of darkness, mRNAs for tubulin and associated flagellar proteins were found to accumulate periodically with a peak just prior to cell division. This was not seen when previously synchronized cells were transferred to constant environmental conditions in a turbidostat, suggesting that dramatic changes in tubulin mRNA levels are not required for successful completion of the cell cycle. A hypothesis to explain the patterns of tubulin mRNA accumulation found under different environmental conditions is presented.

Patterns of protein synthesis during the cell cycle of the fission yeast Schizosaccharomyces pombe

1982 ◽  
Vol 58 (1) ◽  
pp. 263-285
Author(s):  
J. Creanor ◽  
J.M. Mitchison
Keyword(s):  
Cell Cycle ◽  
Protein Synthesis ◽  
Schizosaccharomyces Pombe ◽  
Messenger Rna ◽  
Model Fitting ◽  
Selection Procedure ◽  
Periodic Pattern ◽  
Synchronous Cultures ◽  
Metabolic Pool ◽  
Cdc 2

The rate of protein synthesis through the cell cycle of Schizosaccharomyces pombe has been determined from the incorporation of pulses of [3H]tryptophan in synchronous cultures prepared by selection in an elutriating rotor. This selection procedure caused minimal perturbations as judged by asynchronous control cultures, which had also been put through the rotor. The rate of synthesis showed a periodic pattern rather than a smooth exponential increase. There was a sharp increase in the rate at an ‘acceleration point’ at about 0.9 of the cycle. Model-fitting by a novel procedure suggests that the average single cell has an increasing rate of protein synthesis for the first 60% of the cycle and a constant rate for the remaining 40%. The same pattern was shown in less extensive experiments with [3H]leucine and [3H]phenylalanine. It was also shown in a series of size mutants, which indicates that the pattern is not size-related, in contrast to earlier work on the rates of synthesis of messenger RNA. However, one large mutant (cdc 2.M35r20) had a significantly earlier acceleration point. Care was taken to justify the assumption that the rate of incorporation of tryptophan was a valid measure of the rate of protein synthesis. A tryptophan auxotroph was used to eliminate the problem of endogenous supply and the size of the metabolic pool was measured through the cycle. This pool did not show cell-cycle related fluctuations. An operational model of the pools is presented.

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