scholarly journals Effect of drugs on deoxyribonucleic acid synthesis in isolated mammalian cell nuclei. Comparison with partially purified deoxyribonucleic acid polymerases

1979 ◽  
Vol 178 (3) ◽  
pp. 621-626 ◽  
Author(s):  
J F Burke ◽  
P M Duff ◽  
C K Pearson
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
Cell Nuclei ◽  
Isolated Nuclei ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta ◽  
Deoxyribonucleic Acid Synthesis

In order to ascertain the identity of the DNA-dependent DNA polymerase responsible for the observed DNA synthesis in nuclei isolated from baby-hamster kidney (BHK-21/C13) cells a comparative study was carried out on the effects of some drugs, reported to influence DNA synthesis, on DNA synthesis catalysed by these nuclei and by partially purified DNA polymerase-alpha and -beta. In all cases DNA synthesis by isolated nuclei and polymerase-alpha was inhibited to similar extents by N-ethylmaleimide, p-hydroxymercuribenzoate, novobiocin, heparin and phosphonoacetic acid; polymerase-beta was much less affected by these compounds. Ethidium bromide inhibited all DNA synthesis to similar extents, although at low concentrations (about 2 microgram/ml) synthesis in isolated nuclei was stimulated. The results are discussed in relation to the proposal that DNA polymerase-alpha catalyses the covalent extension of Okazaki fragments that these nuclei carry out in vitro.

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.

scholarly journals Deoxyribonucleic acid synthesis in mammalian nuclei. Incorporation of deoxyribonucleotides and chain-terminating nucleotide analogues

1971 ◽  
Vol 121 (5) ◽  
pp. 803-809 ◽  
Author(s):  
M. A. Waqar ◽  
L. A. Burgoyne ◽  
M. R. Atkinson
Keyword(s):  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
Deoxyribonucleic Acid Synthesis ◽  
Dna Chain ◽  
Relationship Of ◽  
The Relationship ◽  
Neighbour Analysis

The properties of a nuclear preparation from rat liver and thymus are described. (1) Nearest-neighbour analysis after incorporation of 32P-labelled nucleotide residues from dATP, dCTP, dGTP, dTTP and arabinofuranosyl analogues of CTP and ATP shows template-dependent DNA synthesis. (2) Where primer termini are limiting, incorporation of arabinofuranosyl analogues of AMP and CMP residues proceeds to a limit indicating that both of these analogues are DNA chain terminators. (3) No large differences have been found between the priming potentialities or the intrinsic DNA polymerase activities of nuclei from resting or regenerating liver and the relationship of this DNA synthesis in vitro to DNA replication or repair in vivo is briefly discussed.

USE OF I125-LABELED 5-IODO-2′-DEOXYURIDINE FOR THE MEASUREMENT OF DNA SYNTHESIS IN MAMMALIAN CELLS IN VITRO

1963 ◽  
Vol 41 (11) ◽  
pp. 2343-2351 ◽  
Author(s):  
S. Mak ◽  
J. E. Till
Keyword(s):  
Dna Synthesis ◽  
Mammalian Cells ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
X Rays ◽  
Beta Particles ◽  
Deoxyribonucleic Acid Synthesis ◽  
Gamma Emitter

The use of isotopically labeled 5-iodo-2′-deoxyuridine (I125UdR) for determination of the rate of deoxyribonucleic acid synthesis in mammalian cells in vitro has been investigated. The results obtained indicate that for this purpose I125UdR is a suitable substitute for the more commonly used DNA precursor, tritium-labeled thymidine (H3TdR). I125UdR appears to be incorporated specifically into the DNA of cells in culture, and has been demonstrated to compete with H3TdR, although the Km for H3TdR was smaller than that of I125UdR by a factor of approximately 4. The amount of label incorporated into DNA of cells increased linearly with time. When the rate of DNA synthesis was reduced by exposure of the cells to various doses of X-rays, the ratio of I125UdR incorporation to H3TdR incorporation into DNA of cells was found to be a constant, which supports the view that uptake of the analogue provides as reliable an indication of effects upon the rate of DNA synthesis as does that of H3TdR. The chief advantage of I125UdR over H3TdR is that I125 is a gamma emitter, so that the difficulties encountered in detection of the low energy beta particles from H3 may be avoided.

scholarly journals The effect of nitrofurantoin on deoxyribonucleic acid synthesis in plant mitochondria

1980 ◽  
Vol 186 (1) ◽  
pp. 325-329 ◽  
Author(s):  
S Palit ◽  
B B Goswami ◽  
D K Dube
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Plant Mitochondria ◽  
Acid Synthesis ◽  
Dna Polymerase I ◽  
Avian Myeloblastosis Virus ◽  
Deoxyribonucleic Acid Synthesis ◽  
Etiolated Seedlings ◽  
Polymerase I

Nitrofurantoin (1-([(5-nitrofuran-2-yl)methylene]amino)imidazolidine-2,4-dione), a widely used drug and also a well-known bacterial mutagen, inhibits DNA synthesis in mitochondria from 48 h etiolated seedlings of Vigna sinensis (Linn.) Savi (snake bean). The effect appears at the level of the uptake of radioactive deoxynucleoside triphosphates by the plant mitochondria. Nitrofurantoin does not inhibit DNA synthesis in vitro by homogeneous Escherichia coli DNA polymerase I and DNA polymerase from avian-myeloblastosis virus. No specific nitroreductase activity could be detected in mitochondria.

scholarly journals TRANSIENT PHOSPHORYLATION OF DEOXYRIBOSIDES AND REGULATION OF DEOXYRIBONUCLEIC ACID SYNTHESIS

1961 ◽  
Vol 11 (2) ◽  
pp. 311-319 ◽  
Author(s):  
Yasuo Hotta ◽  
Herbert Stern
Keyword(s):  
Dna Synthesis ◽  
Enzyme Induction ◽  
Deoxyribonucleic Acid ◽  
Lilium Longiflorum ◽  
Acid Synthesis ◽  
Deoxyribonucleic Acid Synthesis ◽  
Patterns Of Behavior

Microspores isolated from Lilium longiflorum and Trillium erectum were studied with respect to their capacities for phosphorylating deoxyribosides in vitro. It was found that such capacities are manifest only during brief intervals of time adjacent to periods of DNA synthesis, and that none of the neighboring cells in the anther acquire them. The observed patterns of behavior are interpreted in terms of enzyme induction as a device for regulating DNA synthesis.

scholarly journals c-myc protein and DNA replication: separation of c-myc antibodies from an inhibitor of DNA synthesis.

1987 ◽  
Vol 7 (12) ◽  
pp. 4594-4598 ◽  
Author(s):  
C Gutierrez ◽  
Z S Guo ◽  
J Farrell-Towt ◽  
G Ju ◽  
M L DePamphilis
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Isolated Nuclei ◽  
Dna Polymerase Alpha ◽  
Inhibition Of Dna Synthesis ◽  
Antibody Preparations

Antibodies against human c-myc protein have been reported to inhibit DNA polymerase activity and endogenous DNA synthesis in isolated nuclei, suggesting a role for c-myc in DNA replication. Using the same antibody preparations, we observed equivalent inhibition of simian virus 40 DNA replication and DNA polymerase alpha and delta activities in vitro, as well as inhibition of DNA synthesis in isolated nuclei. However, the c-myc antibodies could be completely separated from the DNA synthesis inhibition activity. c-myc antibodies prepared in other laboratories also did not interfere with initiation of simian virus 40 DNA replication, DNA synthesis at replication forks, or DNA polymerase alpha or delta activity. Therefore, the previously reported inhibition of DNA synthesis by some antibody preparations resulted from the presence of an unidentified inhibitor of DNA polymerases alpha and delta and not from the action of c-myc antibodies.

USE OF I125-LABELED 5-IODO-2′-DEOXYURIDINE FOR THE MEASUREMENT OF DNA SYNTHESIS IN MAMMALIAN CELLS IN VITRO

1963 ◽  
Vol 41 (1) ◽  
pp. 2343-2351 ◽  
Author(s):  
S. Mak ◽  
J. E. Till
Keyword(s):  
Dna Synthesis ◽  
Mammalian Cells ◽  
Deoxyribonucleic Acid ◽  
Acid Synthesis ◽  
X Rays ◽  
Beta Particles ◽  
Deoxyribonucleic Acid Synthesis ◽  
Gamma Emitter

The use of isotopically labeled 5-iodo-2′-deoxyuridine (I125UdR) for determination of the rate of deoxyribonucleic acid synthesis in mammalian cells in vitro has been investigated. The results obtained indicate that for this purpose I125UdR is a suitable substitute for the more commonly used DNA precursor, tritium-labeled thymidine (H3TdR). I125UdR appears to be incorporated specifically into the DNA of cells in culture, and has been demonstrated to compete with H3TdR, although the Km for H3TdR was smaller than that of I125UdR by a factor of approximately 4. The amount of label incorporated into DNA of cells increased linearly with time. When the rate of DNA synthesis was reduced by exposure of the cells to various doses of X-rays, the ratio of I125UdR incorporation to H3TdR incorporation into DNA of cells was found to be a constant, which supports the view that uptake of the analogue provides as reliable an indication of effects upon the rate of DNA synthesis as does that of H3TdR. The chief advantage of I125UdR over H3TdR is that I125 is a gamma emitter, so that the difficulties encountered in detection of the low energy beta particles from H3 may be avoided.

c-myc protein and DNA replication: separation of c-myc antibodies from an inhibitor of DNA synthesis

1987 ◽  
Vol 7 (12) ◽  
pp. 4594-4598
Author(s):  
C Gutierrez ◽  
Z S Guo ◽  
J Farrell-Towt ◽  
G Ju ◽  
M L DePamphilis
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Isolated Nuclei ◽  
Dna Polymerase Alpha ◽  
Inhibition Of Dna Synthesis ◽  
Antibody Preparations

Antibodies against human c-myc protein have been reported to inhibit DNA polymerase activity and endogenous DNA synthesis in isolated nuclei, suggesting a role for c-myc in DNA replication. Using the same antibody preparations, we observed equivalent inhibition of simian virus 40 DNA replication and DNA polymerase alpha and delta activities in vitro, as well as inhibition of DNA synthesis in isolated nuclei. However, the c-myc antibodies could be completely separated from the DNA synthesis inhibition activity. c-myc antibodies prepared in other laboratories also did not interfere with initiation of simian virus 40 DNA replication, DNA synthesis at replication forks, or DNA polymerase alpha or delta activity. Therefore, the previously reported inhibition of DNA synthesis by some antibody preparations resulted from the presence of an unidentified inhibitor of DNA polymerases alpha and delta and not from the action of c-myc antibodies.

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