scholarly journals MICRONUCLEAR RNA SYNTHESIS IN PARAMECIUM CAUDATUM

1967 ◽  
Vol 33 (2) ◽  
pp. 281-285 ◽  
Author(s):  
M. V. Narasimha Rao ◽  
David M. Prescott

In a generation time of 8 hr in Paramecium caudatum, the bulk of DNA synthesis detected by thymidine-3H incorporation takes place in the latter part of the cell cycle. The micronuclear cycle includes a G1 of 3 hr followed by an S period of 3–3½ hr. G2 and division occupies the remaining period of the cycle. Macronuclear RNA synthesis detected by 5'-uridine-3H incorporation is continuous throughout the cell cycle. Micronuclear RNA synthesis is restricted to the S period. Ribonuclease removes 80–90% of the incorporated label. Pulse-chase experiments showed that part of the RNA is conserved and released to the cytoplasm during the succeeding G1 period.

1974 ◽  
Vol 61 (3) ◽  
pp. 591-598 ◽  
Author(s):  
Joan Smith-Sonneborn ◽  
Michael Klass

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.


Blood ◽  
1972 ◽  
Vol 39 (4) ◽  
pp. 575-580 ◽  
Author(s):  
E. Fred Saunders

Abstract The effect of L-asparaginase on the cell cycle and nucleic acid synthesis of leukemic cells was studied in five children with acute lymphoblastic leukemia. Following an intravenous infusion of the drug, serial marrow samples were obtained for buffy coat volume, mitotic index, and autoradiographic assessment of DNA and RNA synthesis using tritiated thymidine and tritiated uridine, respectively. A rapid decline in buffy coat volume indicated a lytic effect on lymphoblasts. There was a greater kill of proliferative (blasts in the cell cycle) than nonproliferative (G0) leukemic cells. Mitotic indices changed little until 24 hr; in contrast, thymidine labeling indices decreased markedly to less than 50% of control by 6 hr. The changes in labeling indices prior to changes in mitotic indices indicated that L-asparaginase blocked the entrance of cells into the DNA synthesis period of the cell cycle. Cells already in DNA synthesis appeared to continue into mitosis. Uridine labeling indices decreased progressively in all patients. Uridine uptake was inhibited equally in both proliferative and nonproliferative blasts. Therefore, inhibition of RNA synthesis by L-asparaginase was independent of the proliferative activity of the marrow.


1984 ◽  
Vol 4 (12) ◽  
pp. 2858-2864 ◽  
Author(s):  
R K Storms ◽  
R W Ord ◽  
M T Greenwood ◽  
B Mirdamadi ◽  
F K Chu ◽  
...  

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.


1975 ◽  
Vol 18 (1) ◽  
pp. 27-39
Author(s):  
H. Fouquet ◽  
R. Bohme ◽  
R. Wick ◽  
H.W. Sauer ◽  
K. Scheller

Hydroxyurea, at concentrations of 40–60 mM, selectively and effectively blocked incorporation of thymidine into DNA. Inhibition occurred within 5–10 min of application of the agent when DNA synthesis was in progress, while the onset of replication at the beginning of S-phase and DNA synthesis in G2 phase were not affected. Uridine incorporation into TCA-precipitable material, in the presence of hydroxyurea, was significantly (up to 70%) inhibited in early S-phase of the cell cycle. Selective inhibition of RNA synthesis was confirmed for RNA separated into rRNA-rich and poly(A)-rich RNA fractions and analysed by the 2 kinds of DNA-RNA hybridization reactions. Uridine incorporation into poly (A) RNA was also inhibited under conditions where cycloheximide prevented maturation of nascent DNA molecules in early S-phase. We assume that chromatin which is replicating early DNA sequences may be a more competent template for transcription.


1976 ◽  
Vol 21 (3) ◽  
pp. 497-521
Author(s):  
R.S. Fraser ◽  
F. Moreno

The rates of polyadenylated messenger RNA and ribosomal RNA synthesis were measured in synchronously dividing cultures of fission yeast (Schizosaccharomyces pombe). Control asynchronous cultures, which had been exposed to the conditions used for preparing synchronous cultures, were investigated to check for effects of the synchronization procedure itself on RNA synthesis. After each period of DNA synthesis in synchronous culture, the rates of messenger and ribosomal RNA synthesis doubled, suggesting that gene number controls the rate of messenger and ribosomal RNA synthesis. This was confirmed by experiments with asynchronous, exponential-phase cultures in which DNA synthesis was inhibited by hydroxyurea. Both synchronous culture and hydroxyurea experiments suggested that there is a delay of 15 min (0-1 of the cell generation time) between replication of the DNA and transcription of both gene copies. A pattern of protein accumulation was calculated from changes in the rate of polyadenylated messenger RNA synthesis during synchronous culture. The simulated pattern indicates that protein is accumulated linearly, with a doubling in the rate of accumulation once per cell cycle. The simulated pattern of protein accumulation is very similar to measurements previously reported by other workers of changes in activities of 3 enzymes in synchronous cultures. It is suggested that the doubling of the rate of messenger RNA synthesis, as a consequence of the replication of the DNA once per cycle, provides the basis of a mechanism for control of the doubling of other cellular constituents during the cell cycle.


1973 ◽  
Vol 59 (3) ◽  
pp. 615-623 ◽  
Author(s):  
P. R. Gabe ◽  
L. E. de Bault

Macromolecular syntheses in encysted Tetrahymena patula were studied using Feulgen fluorescence cytophotometry, autoradiography, and inhibitors of RNA and protein synthesis. Cycloheximide significantly depressed protein synthesis and D-actinomycin effectively blocked RNA synthesis. Under these conditions, the cells within the cyst were unable to divide. Both cytophotometric measurements and autoradiographic data with tritiated thymidine show that DNA synthesis does not occur during the encystment divisions. Excysted cells placed in nutrient broth medium showed a prolonged generation time after the first cell growth cycle, and by the third generation the mean DNA content per cell was almost triple that of starved excysted cells. These findings indicate that (a) the encystment divisions require RNA and protein synthesis, which are apparently effected through turnover, (b) the encystment division cycles occur in the absence of DNA synthesis, and (c) excysted cells placed in culture medium may go through more than one DNA replication per cell cycle.


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

1973 ◽  
Vol 59 (1) ◽  
pp. 1-11 ◽  
Author(s):  
William R. Jeffery ◽  
Joseph Frankel ◽  
Lawrence E. de Bault ◽  
Leslie M. Jenkins

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.


Development ◽  
1973 ◽  
Vol 29 (3) ◽  
pp. 745-751
Author(s):  
D. B. Wilson

Radioautographic data obtained from a total of 103 chick embryos injected with [3H]thymidine suggest that the generation time of neuroepithelial cells in dorsolateral regions of the optic tectum increases from approximately 8 h at 3 days of incubation to 15 h at 6 days of incubation. The most marked change occurs between the 4th and 5th day, when the generation time increases from 9 to 13 h, respectively. Between the 3rd and 6th day of incubation the length of the DNA-synthetic (S) period and of the premitotic (G2) period remains fairly constant; however, the duration of mitosis (M) and of the postmitotic (G1) period appears to be prolonged.


1984 ◽  
Vol 4 (12) ◽  
pp. 2858-2864
Author(s):  
R K Storms ◽  
R W Ord ◽  
M T Greenwood ◽  
B Mirdamadi ◽  
F K Chu ◽  
...  

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.


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