scholarly journals THE SYNTHESIS OF DNA, RNA, AND NUCLEAR PROTEIN IN NORMAL AND TUMOR STRAIN CELLS

1966 ◽  
Vol 28 (2) ◽  
pp. 263-275 ◽  
Author(s):  
John Seed
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Hela Cells ◽  
Nuclear Protein ◽  
X Rays ◽  
Normal Cells ◽  
Human Carcinoma ◽  
Physical Measurements ◽  
Cytoplasmic Rna ◽  
Nuclear Rna

Interferometric and photometric measurements have been made on HeLa cells, a strain of cells originally derived from a human carcinoma. From a study of the relations between successive physical measurements on individual cells, it was confirmed that, whereas the net syntheses of nuclear RNA and nuclear protein are closely associated during interphase, they are dissociated from DNA replication to a significant extent. These results on nuclear metabolism agree with others previously reported in cell strains derived from tumors; they contrast with results from freshly prepared normal cells, where the net syntheses of DNA, nuclear RNA, and protein are closely associated during interphase. Cytoplasmic measurements on HeLa cells showed that much of the net synthesis of cytoplasmic RNA is associated with DNA replication as in normal cells, and they failed to detect transfer from the nucleus of a stable RNA component synthesized independently from DNA replication. In auxiliary experiments, an inhibition of the onset of DNA synthesis was produced by a dose of X-rays; under these conditions it was shown that the major part of the accumulation of nuclear protein was independent of DNA replication and that the accumulation of nuclear RNA was equivalent to or slightly less than that of nuclear protein. About half the accumulation of cytoplasmic RNA was inhibited when DNA synthesis was blocked.

scholarly journals THE SYNTHESIS OF DNA, RNA, AND NUCLEAR PROTEIN IN NORMAL AND TUMOR STRAIN CELLS

1966 ◽  
Vol 28 (2) ◽  
pp. 249-256 ◽  
Author(s):  
John Seed
Keyword(s):  
Dna Synthesis ◽  
Cell Cultures ◽  
Nuclear Protein ◽  
Mouse Cell ◽  
X Rays ◽  
Normal Tissues ◽  
Physical Measurements ◽  
Cytoplasmic Rna ◽  
Nuclear Rna ◽  
Photometric Measurements

Interferometric and photometric measurements have been made on replicating embryo mouse cell cultures. From a study of the relations between successive physical measurements on individual cells, it was found that the net syntheses of DNA, nuclear RNA, nuclear protein, and cytoplasmic RNA are closely associated during interphase. In auxiliary experiments, an inhibition of the onset of DNA synthesis (produced by a dose of X-rays) was found to block the majority of the accumulation of nuclear protein and nuclear RNA. These results are consistent with others previously reported in dividing cell cultures freshly prepared from normal tissues.

scholarly journals THE SYNTHESIS OF DNA, RNA, AND NUCLEAR PROTEIN IN NORMAL AND TUMOR STRAIN CELLS

1966 ◽  
Vol 28 (2) ◽  
pp. 233-248 ◽  
Author(s):  
John Seed
Keyword(s):  
Cell Cultures ◽  
Nuclear Protein ◽  
Kidney Cells ◽  
X Rays ◽  
Normal Tissues ◽  
Physical Measurements ◽  
Cytoplasmic Rna ◽  
Human Cell Cultures ◽  
Nuclear Rna ◽  
Photometric Measurements

Interferometric and photometric measurements have been made on replicating embryo human cell cultures. From a study of the relations between successive physical measurements on individual cells, it was found that the net syntheses of DNA, nuclear RNA, nuclear protein, and cytoplasmic RNA are closely associated during interphase. Additional measurements of DNA and cytoplasmic RNA on freshly prepared replicating monkey kidney cells gave similar results. In auxiliary experiments with embryo human cells, an inhibition of the onset of DNA synthesis (produced by a dose of X-rays) was found to block the majority of the accumulation of nuclear protein and RNA and about half the accumulation of cytoplasmic RNA. These results are consistent with others previously reported in dividing cell cultures freshly prepared from normal tissues.

scholarly journals THE SYNTHESIS OF DNA, RNA, AND NUCLEAR PROTEIN IN NORMAL AND TUMOR STRAIN CELLS

1966 ◽  
Vol 28 (2) ◽  
pp. 257-261 ◽  
Author(s):  
John Seed
Keyword(s):  
Dna Synthesis ◽  
Nuclear Protein ◽  
Ascites Tumor ◽  
Significant Extent ◽  
Physical Measurements ◽  
Nuclear Rna ◽  
X Irradiation ◽  
Mouse Ascites ◽  
Cell Strains ◽  
Photometric Measurements

Interferometric and photometric measurements have been made on replicating mouse ascites tumor cell cultures. From a study of the relations between successive physical measurements on individual cells, it was found that whereas the net syntheses of nuclear RNA and nuclear protein are closely associated during interphase, they are dissociated from DNA replication to a significant extent. These results agree with others reported in replicating cell strains derived from tumors. In auxiliary experiments an attempt was made to block the initiation of DNA synthesis by X-irradiation: although large amounts of nuclear protein accumulated in some cells in the absence of DNA synthesis, the inability to hold the DNA block for an interphase time prevented a quantitative analysis of the results.

A human nuclear protein with sequence homology to a family of early S phase proteins is required for entry into S phase and for cell division

1994 ◽  
Vol 107 (1) ◽  
pp. 253-265 ◽  
Author(s):  
I.T. Todorov ◽  
R. Pepperkok ◽  
R.N. Philipova ◽  
S.E. Kearsey ◽  
W. Ansorge ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Amino Acid ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Mammalian Cells ◽  
Nuclear Protein ◽  
Amino Acid Level ◽  
S Phase ◽  
Significant Similarity

Molecular cloning and characterisation of a human nuclear protein designated BM28 is reported. On the amino acid level this 892 amino acid protein, migrating on SDS-gels as a 125 kDa polypeptide, shares areas of significant similarity with a recently defined family of early S phase proteins. The members of this family, the Saccharomyces cerevisiae Mcm2p, Mcm3p, Cdc46p/Mcm5p, the Schizosaccharomyces pombe Cdc21p and the mouse protein P1 are considered to be involved in the onset of DNA replication. The highest similarity was found with Mcm2p (42% identity over the whole length and higher than 75% over a conservative region of 215 amino acid residues), suggesting that BM28 could represent the human homologue of the S. cerevisiae MCM2. Using antibodies raised against the recombinant BM28 the corresponding antigen was found to be localised in the nuclei of various mammalian cells. Microinjection of anti-BM28 antibody into synchronised mouse NIH3T3 or human HeLa cells presents evidence for the involvement of the protein in cell cycle progression. When injected in G1 phase the anti-BM28 antibody inhibits the onset of subsequent DNA synthesis as tested by the incorporation of bromodeoxyuridine. Microinjection during the S phase had no effect on DNA synthesis, but inhibits cell division. The data suggest that the nuclear protein BM28 is required for two events of the cell cycle, for the onset of DNA replication and for cell division.

Acid-Soluble Deoxynucleotides and DNA Synthesis in Growing Yeast After X-Irradiation, I. Diphenylamine Positive Material in Synchronized and Asynchronously Growing Cells

1971 ◽  
Vol 26 (12) ◽  
pp. 1266-1270
Author(s):  
Hans Eckstein
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Normal Values ◽  
X Rays ◽  
Considerable Decrease ◽  
Positive Material ◽  
Normal Degree ◽  
X Irradiation ◽  
Diphenylamine Reaction

In synchronized growing yeast, the level of acid-soluble purine deoxynucleotides, as determined by the diphenylamine reaction, fluctuates rhythmically: A rapid increase each time before DNA starts to replicate is followed by a considerable decrease during DNA augmentation. Delay of DNA replication by irradiation of synchronized yeast with 50 kr of X-rays results in a stepwise augmentation of deoxyribose derivatives. The fluctuating behaviour is restored, when DNA starts to increase, again. During asynchronous growth acid-soluble deoxyribosidic compounds are augmented with a considerably increased rate, when DNA replication is delayed by X-rays. The resulting unusual pool seize is maintained, even if DNA replication is resumed to a normal degree, but the rate of further increase is reduced to normal values. Parts of the rapidly accumulated deoxyribosidic material differ with some properties from purine deoxynucleotides.

scholarly journals Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 K-bZIP Modulates Latency-Associated Nuclear Protein-Mediated Suppression of Lytic Origin-Dependent DNA Synthesis

2009 ◽  
Vol 83 (17) ◽  
pp. 8492-8501 ◽  
Author(s):  
Cyprian Rossetto ◽  
Irena Yamboliev ◽  
Gregory S. Pari
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Nuclear Protein ◽  
Human Herpesvirus ◽  
Lytic Replication ◽  
Human Herpesvirus 8 ◽  
Dependent Manner ◽  
Herpesvirus 8 ◽  
Replication Assay ◽  
Lytic Dna Replication

ABSTRACT The original cotransfection replication assay identified eight human herpesvirus 8 (HHV8)-encoded proteins required for origin-dependent lytic DNA replication. Previously, we demonstrated that under conditions where K-Rta is overexpressed, a K-bZIP knockout bacmid displayed an aberrant subcellular localization pattern for the latency-associated nuclear protein (LANA). Additionally, these same studies demonstrated that K-bZIP interacts with LANA in the absence of K-Rta and that K-bZIP does not directly participate in, but may facilitate, the initiation of lytic DNA synthesis. We developed a modification of the transient cotransfection replication assay wherein both lytic (oriLyt) and latent (terminal repeat) DNA replication are evaluated simultaneously. We now show that LANA represses origin-dependent lytic DNA replication in a dose dependent manner when added to the cotransfection replication assay. This repression was overcome by increasing amounts of a K-bZIP expression plasmid in the cotransfection mixture or by dominant-negative inhibition of the interaction of LANA with K-bZIP by the overexpression of the K-bZIP-LANA binding domain. Chromatin immunoprecipitation assays show that LANA interacts with oriLyt in the absence of K-bZIP expression, suggesting that suppression of lytic replication by LANA is mediated by direct binding. The interaction of K-bZIP with oriLyt was dependent upon the expression of LANA; however, LANA interacted with oriLyt independently of K-bZIP expression. These data suggest that the interaction of LANA with K-bZIP modulates lytic and latent replication and that K-bZIP facilitates lytic DNA replication and modulates the switch from the latent phase of the virus.

scholarly journals Aphidicolin does not inhibit DNA repair synthesis in ultraviolet-irradiated HeLa cells. A radioautographic study

1981 ◽  
Vol 199 (2) ◽  
pp. 453-455 ◽  
Author(s):  
N Hardt ◽  
G Pedrali-Noy ◽  
F Focher ◽  
S Spadari
Keyword(s):  
Dna Repair ◽  
Dna Replication ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Hela Cells ◽  
Nuclear Dna ◽  
Detectable Effect ◽  
Repair Synthesis ◽  
Dna Polymerase Alpha ◽  
Dna Repair Synthesis

A radioautographic examination of nuclear DNA synthesis in unirradiated and u.v.-irradiated HeLa cells, in the presence and in the absence of aphidicolin, showed that aphidicolin inhibits nuclear DNA replication and has no detectable effect on DNA repair synthesis. Although the results establish that in u.v.-irradiated HeLa cells most of the DNA repair synthesis is not due to DNA polymerase alpha, they do not preclude a significant role for this enzyme in DNA repair processes.

scholarly journals Differences in sequence content of nuclear and cytoplasmic polyribosomal RNA from adenovirus-infected cells

1984 ◽  
Vol 218 (2) ◽  
pp. 583-590
Author(s):  
N K Chatterjee ◽  
C Tuchowski ◽  
G E Eagan ◽  
T M Haley
Keyword(s):  
Hela Cells ◽  
Sucrose Gradient ◽  
Actinomycin D ◽  
Rna Molecules ◽  
Filter Hybridization ◽  
Cytoplasmic Rna ◽  
Blot Technique ◽  
Nuclear Rna ◽  
Infected Cells ◽  
Two Populations

RNA molecules from nuclear and cytoplasmic polyribosomes of adenovirus-infected HeLa cells were compared by hybridization to analyse the sequence content. Nuclear polyribosomes were released by exposure of intact detergent-washed nuclei to poly(U) and purified. Cytoplasmic polyribosomes were also purified from the same cells. To show that nuclear polyribosomes contain ribosomes linked by mRNA, polyribosomes were labelled with methionine and uridine in the presence of actinomycin D in adenovirus-infected cells. Purified nuclear polyribosomes were treated with EDTA under conditions which dissociate polyribosomes into ribosomes and subunits with a simultaneous release of mRNA, and sedimented. The treatment dissociated these polyribosomes, releasing the mRNA from them. Radiolabelled total RNA from each polyribosome population was fractionated in sucrose gradients into several pools or hybridized to intact adenovirus DNA to select virus-specific RNA. Sucrose-gradient-fractionated pool-3 RNA (about 28S) and virus-specific RNA were then hybridized to fragments of adenovirus DNA cleaved by restriction endonucleases SmaI, HindIII and EcoRI by the Southern-blot technique and by filter hybridization. The results showed that nuclear RNA contained sequences, from about 0 to 18 map units, which were essentially absent from cytoplasmic RNA. Furthermore, the amount of virus-specific RNA for a particular sequence was also different in the two populations.

Wirkung von Lithium- und Rhodanidionen auf den Nukleinsäure-Stoffwechsel von Tetrahymena pyriformis, normalen und neoplastischen Säugerzellen / Effect of Lithium and Thiocyanate Ions on the Metabolism of Nucleic Acids of Tetrahymena pyriformis, Normal and Neoplastic Mammalian Cells

1973 ◽  
Vol 28 (7-8) ◽  
pp. 460-462 ◽  
Author(s):  
Klaus Wayss ◽  
Manfred Volm
Keyword(s):  
Nucleic Acids ◽  
Dna Synthesis ◽  
Tumor Cells ◽  
Hela Cells ◽  
Mammalian Cells ◽  
Tetrahymena Pyriformis ◽  
Ascites Tumor ◽  
Yoshida Sarcoma ◽  
Normal Cells ◽  
I Cells

An antagonistic influence of lithium and thiocyanate ions on the metabolism of nucleic acids has been demonstrated in Tetrahymena pyriformis. This effect corresponds to morphogenetic observations on other ciliates. Similar investigations on mammalian cells (L-cells, CV I-cells, Chang-livercells, HeLa-cells, and ascites tumor cells of Walker-carcinosarcoma 256, Yoshida-sarcoma, Zajdelahepatoma) showed, in contrast to the results in Tetrahymena, no antagonistic influence of lithium and thiocyanate ions: Both ions inhibit the DNA-synthesis, in tumor cells as well as in normal cells.

The supply of exogenous deoxyribonucleotides accelerates the speed of the replication fork in early S-phase

2001 ◽  
Vol 114 (4) ◽  
pp. 747-750
Author(s):  
J. Malinsky ◽  
K. Koberna ◽  
D. Stanek ◽  
M. Masata ◽  
I. Votruba ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Hela Cells ◽  
Replication Fork ◽  
S Phase ◽  
Limiting Factor ◽  
Replication Forks ◽  
Replication Process ◽  
Average Speed ◽  
Rate Limiting

Earlier studies have established that the average speed of a replication fork is two to three times slower in early S-phase than in late S-phase and that the intracellular 2′-deoxyribonucleoside 5′-triphosphate pools grow during S-phase. In this study, the effect of the exogenous 2′-deoxyribonucleoside 5′-triphosphate (dNTP) supply on the average replication speed in a synchronised population of human HeLa cells was tested. The speed of replication fork movement was measured on extended DNA fibers labelled with 2′-deoxythymidine analogues 5-chloro-2′-deoxyuridine and 5-iodo-2′-deoxyuridine. We show that the introduction of exogenous dNTPs accelerates the replication process at the beginning of DNA synthesis only. In late S-phase, the administration of additional dNTPs has no effect on the speed of replication forks. The availability of 2′-deoxynucleotides seems to be a rate-limiting factor for DNA replication during early S-phase.

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