scholarly journals Microbubbles in replicating nuclear deoxyribonucleic acid from Physarum polycephalum

1979 ◽  
Vol 183 (2) ◽  
pp. 477-480 ◽  
Author(s):  
D A F Gillespie ◽  
N Hardman
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Dna Synthesis ◽  
Physarum Polycephalum ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
S Phase ◽  
Dna Structures ◽  
Nuclear Deoxyribonucleic Acid

Clusters of microbubbles, represent probable sites of newly initiated DNA synthesis, were identified in nuclear DNA from Physarum polycephalum by using the electron microscope. Their presence is associated specifically with S-phase. Each microbubble corresponds in size to a replicating segment of DNA about 100-5000 nucleotide residues in length. The DNA structures containing microbubbles are metastable, and revert to native DNA in the presence of moderate concentrations of formamide used to prepare samples for electron microscopy. It is suggested that each cluster of microbubbles may correspond to a unit of replication (a replicon) in Physarum DNA.

scholarly journals LIMITED DNA SYNTHESIS IN THE ABSENCE OF PROTEIN SYNTHESIS IN PHYSARUM POLYCEPHALUM

1966 ◽  
Vol 31 (3) ◽  
pp. 577-583 ◽  
Author(s):  
J. E. Cummins ◽  
H. P. Rusch
Keyword(s):  
Protein Synthesis ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Physarum Polycephalum ◽  
Nuclear Dna ◽  
S Phase ◽  
Early Prophase ◽  
Late Prophase ◽  
Removal Experiments ◽  
Inhibitor Of Protein Synthesis

Actidione (cycloheximide), an antibiotic inhibitor of protein synthesis, blocked the incorporation of leucine and lysine during the S phase of Physarum polycephalum. Actidione added during the early prophase period in which mitosis is blocked totally inhibited the initiation of DNA synthesis. Actidione treatment in late prophase, which permitted mitosis in the absence of protein synthesis, permitted initiation of a round of DNA replication making up between 20 and 30% of the unreplicated nuclear DNA. Actidione treatment during the S phase permitted a round of replication similar to the effect at the beginning of S. The DNA synthesized in the presence of actidione was replicated semiconservatively and was stable through at least the mitosis following antibiotic removal. Experiments in which fluorodeoxyuridine inhibition was followed by thymidine reversal in the presence of actidione suggest that the early rounds of DNA replication must be completed before later rounds are initiated.

Continuous Nucleolar DNA Synthesis in Late-Interphase Nuclei of Physarum Polycephalum after Transplantation into Postmitotic Plasmodia

1974 ◽  
Vol 15 (1) ◽  
pp. 131-143
Author(s):  
E. GUTTES
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Physarum Polycephalum ◽  
Nuclear Dna ◽  
S Phase ◽  
Interphase Nuclei ◽  
G2 Phase

In the myxomycete, Physarum polycephalum, nuclear DNA synthesis commences immediately upon completion of mitosis. While the synthesis of extranucleolar DNA is completed within a few hours, nucleolar DNA synthesis occurs during most of the S-phase and the entire G2 phase of the intermitotic period. When large (polyploid), late-interphase nuclei were allowed to bypass mitosis by transplantation into recipient plasmodia which were at early interphase and which belonged to a strain having smaller nuclei, the nucleolar DNA of the transplanted nuclei continued to be labelled (autoradiographs) after incubation of the host plasmodia with [3H]thymidine until they entered prophase along with the nuclei of the host plasmodium, approximately one intermitotic period later. This labelling was DNase-sensitive and RNase-resistant. When late-interphase nuclei were labelled with [3H]thymidine just prior to transplantation, there was no decrease of label after transplantation during the additional intermitotic period. We conclude from these experiments that there is no obligatory alternation between nucleolar DNA duplication and mitosis in Physarum polycephalum and that nucleolar DNA replication might exhibit amplification during an experimentally prolonged intermitotic period.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

Continuous DNA replication in the nucleus of the dinoflagellate Prorocentrum micans (Ehrenberg)

1977 ◽  
Vol 27 (1) ◽  
pp. 81-90
Author(s):  
S.A. Filfilan ◽  
D.C. Sigee
Keyword(s):  
Electron Microscope ◽  
Dna Synthesis ◽  
Nuclear Dna ◽  
Continuous Process ◽  
Prorocentrum Micans ◽  
Interphase Nuclei ◽  
Electron Microscope Autoradiography ◽  
Dividing Cells ◽  
High Degree ◽  
Very High

The uptake of tritiated thymine into cells of a heterogeneous population of Prorocentrum micans was investigated using light-microscope and electron-microscope autoradiography. Specificity of thymine uptake into DNA was demonstrated by the specific removal of label from wax-embedded material using DNase and by the high degree of localization of nuclear label to chromosomes in the electron-microscope autoradiographs. All nuclei, including both dividing and non-dividing cells, showed a substantial uptake of label, indicating that nuclear DNA synthesis in Prorocentrum micans is a continuous process. The level of DNA synthesis does show considerable variation, however, with very high levels in some interphase nuclei. The continuous replication of nuclear DNA provides further evidence of dinoflagellate affinity to the prokaryotes, and indicates that Prorocentrum micans is a very primitive eukaryote cell.

scholarly journals MICRONUCLEAR RIBONUCLEIC ACID IN TETRAHYMENA PYRIFORMIS

1970 ◽  
Vol 47 (2) ◽  
pp. 460-467 ◽  
Author(s):  
Kuruganti G. Murti ◽  
David M. Prescott
Keyword(s):  
Electron Microscope ◽  
Dna Synthesis ◽  
Ribonucleic Acid ◽  
Tetrahymena Pyriformis ◽  
S Phase ◽  
Rna Labeling ◽  
Labeled Rna

The presence of RNA in the micronucleus of Tetrahymena pyriformis was detected by electron microscope radioautography after incubation with tritiated precursors. The specificity of RNA labeling was shown by ribonuclease digestion. The period of appearance of labeled RNA in the micronucleus is approximately coincident with the DNA synthesis period for the micronucleus. Pulse-chase experiments showed that the micronuclear RNA disappears during the interphase period. The experiments do not distinguish whether the micronuclear RNA is synthesized in situ or acquired by migration from the macronucleus. In either case it is notable that the appearance of labeled RNA is detected in the micronucleus only during the micronuclear S phase.

scholarly journals The diagnostic significance of nuclear deoxyribonucleic acid measurement in automated cytology.

1979 ◽  
Vol 27 (1) ◽  
pp. 520-521 ◽  
Author(s):  
E Sprenger ◽  
S Witte
Keyword(s):  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Malignant Cell ◽  
Cell Populations ◽  
Diagnostic Significance ◽  
Flow Cytometric ◽  
Screening Systems ◽  
Nuclear Deoxyribonucleic Acid

Flow cytometric analysis of cytologic samples from four different organs shows that nuclear DNA content of malignant cell populations depends to a large extent on organ of origin of the tumor. This fact must be considered in planning screening systems.

scholarly journals Cell population heterogeneity in the inducibility of DNA synthesis in human diploid fibroblasts.

1981 ◽  
Vol 89 (2) ◽  
pp. 194-197 ◽  
Author(s):  
M V Rao
Keyword(s):  
Dna Synthesis ◽  
Cytochalasin B ◽  
Nuclear Dna ◽  
S Phase ◽  
Population Heterogeneity ◽  
Diploid Fibroblasts ◽  
Binucleate Cells ◽  
Human Diploid Fibroblasts ◽  
Significant Difference ◽  
Sister Cells

The initiation of nuclear DNA synthesis has been studied in cytochalasin B (CB)-induced binucleate human diploid fibroblasts (WI-38 cells). Mitotic cells from different passage levels were rendered binucleate by a brief pulse of CB. The cells were then washed free of the drug, and DNA synthesis was studied by [3H]thymidine labeling. The results showed that, in a small percentage of binucleate cells, one nucleus was labeled (S phase) and the other nucleus was unlabeled (G1 phase). There was no significant difference in the percentage of these cells with increasing passage levels. The results of this study suggest that some WI-38 cells retire from the cell cycle at different passage levels, and thereby become refractory to inducers of nuclear DNA synthesis generated by sister cells in S phase.

scholarly journals REGULATION OF DNA REPLICATION IN THE NUCLEI OF THE SLIME MOLD PHYSARUM POLYCEPHALUM

1968 ◽  
Vol 37 (3) ◽  
pp. 761-772 ◽  
Author(s):  
Sophie Guttes ◽  
Edmund Guttes
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Physarum Polycephalum ◽  
Slime Mold ◽  
S Phase ◽  
G2 Phase

Nuclei in G2 phase of the slime mold Physarum polycephalum, when transplanted, by plasmodial coalescence, into an S-phase plasmodium, failed to start another round of DNA synthesis. In the reciprocal combination, S-phase nuclei in a G2-phase host continued DNA synthesis for several hours without appreciable decrease in rate. It is suggested that the beginning of DNA replication is determined by an event, either during or shortly after mitosis, which renders the chromosomes structurally competent for DNA replication.

Some evidence for replication-transcription coupling in Physarum polycephalum

1975 ◽  
Vol 18 (1) ◽  
pp. 27-39
Author(s):  
H. Fouquet ◽  
R. Bohme ◽  
R. Wick ◽  
H.W. Sauer ◽  
K. Scheller
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Dna Sequences ◽  
Physarum Polycephalum ◽  
Rna Synthesis ◽  
Selective Inhibition ◽  
S Phase ◽  
Dna Molecules ◽  
Uridine Incorporation ◽  
G2 Phase

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.

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