Initiation of DNA synthesis in a system of synchronized L-cells: effect of actinomycin D

1968 ◽  
Vol 14 (1) ◽  
pp. 113-122 ◽  
Author(s):  
C. Mittermayer ◽  
P. Kaden ◽  
U. Trommershaeuser ◽  
W. Sandritter
Keyword(s):  
Dna Synthesis ◽  
Actinomycin D ◽  
L Cells

scholarly journals NUCLEAR MEMBRANES OF CULTURED MAMMALIAN CELLS IN THE PERIOD PRECEDING DNA SYNTHESIS

1965 ◽  
Vol 25 (1) ◽  
pp. 103-107 ◽  
Author(s):  
Susumu Kishimoto ◽  
Irving Lieberman
Keyword(s):  
Ionizing Radiation ◽  
Dna Synthesis ◽  
Electrophoretic Mobility ◽  
Mammalian Cells ◽  
Elevated Level ◽  
Actinomycin D ◽  
Kidney Cells ◽  
Nuclear Membranes ◽  
L Cells

When kidney cells are cultured directly from the rabbit, the nuclear membranes undergo a change that can be measured as an increase in electrophoretic mobility. The change appears to begin immediately upon culture and is maximal 2 hours later, after which the mobility remains constant at the elevated level. Actinomycin D and p-fluorophenylalanine, but not EDTA or ionizing radiation, suppress the increase in nuclear electrophoretic mobility. With synchronously growing L cells, no change was detected in nuclei from cells taken during various parts of the division cycle.

scholarly journals Identification of a BALB/c-3T3 cell protein modulated by platelet-derived growth factor.

1983 ◽  
Vol 3 (1) ◽  
pp. 70-81 ◽  
Author(s):  
C D Scher ◽  
R L Dick ◽  
A P Whipple ◽  
K L Locatell
Keyword(s):  
Growth Factor ◽  
Dna Synthesis ◽  
Cell Lines ◽  
Actinomycin D ◽  
Platelet Derived Growth Factor ◽  
Cell Protein ◽  
Transformed Cells ◽  
Antigenic Determinants ◽  
3T3 Cells ◽  
Constitutive Synthesis

The platelet-derived growth factor (PDGF) stimulates density-arrested BALB/c-3T3 cells to synthesize a protein (pII; Mr, 35,000) that is constitutively synthesized by spontaneously transformed BALB/c-3T3 (ST2-3T3) cells which do not require PDGF for growth. Antisera against a major excreted protein family (MEP) of retrovirus-transformed cells quantitatively precipitated cellular pII. PDGF-stimulated pII has the same molecular weight, a similar charge, and similar antigenic determinants as authentic MEP isolated from ST2-3T3 or retrovirus-transformed cells. MEP represented about 2% of the nonnuclear proteins synthesized by ST2-3T3 cells and 0.3 to 0.6% of the proteins synthesized by PDGF-treated BALB/c-3T3 cells, a three- to sixfold increase over the background. In BALB/c-3T3 cells, less PDGF was required for pII (MEP) synthesis than for DNA synthesis. PDGF induced a selective increase in pII (MEP) within 40 min. Such preferential synthesis was inhibited by brief treatment with actinomycin D, suggesting a requirement for newly formed RNA. The constitutive synthesis of pII (MEP) by ST2-3T3 cells was not inhibited by actinomycin D. Five spontaneously or chemical carcinogen-transformed tumorigenic BALB/c-3T3 cell lines were studied; they neither required PDGF for growth nor responded to it. These cell lines became arrested at confluence with a G1 DNA content. Each of these independently isolated lines synthesized pII (MEP) constitutively. Thus, the synthesis of pII (MEP) may be required, but is not sufficient, for PDGF-modulated DNA synthesis.

DNA synthesis in partially synchronized L cells

1962 ◽  
Vol 26 (2) ◽  
pp. 318-326 ◽  
Author(s):  
J.W. Littlefield
Keyword(s):  
Dna Synthesis ◽  
L Cells

scholarly journals The inhibition of nucleic acid synthesis by mycophenolic acid

1969 ◽  
Vol 113 (3) ◽  
pp. 515-524 ◽  
Author(s):  
T J Franklin ◽  
Jennifer M. Cook
Keyword(s):  
Dna Synthesis ◽  
Mycophenolic Acid ◽  
Early Stage ◽  
Acid Synthesis ◽  
Purine Nucleotides ◽  
Free System ◽  
L Cells ◽  
A Cell ◽  
Inhibition Of Dna Synthesis

1. Mycophenolic acid, an antibiotic of some antiquity that more recently has been found to have marked activity against a range of tumours in mice and rats, strongly inhibits DNA synthesis in the L strain of fibroblasts in vitro. 2. The extent of the inhibition of DNA synthesis is markedly increased by preincubation of the cells with mycophenolic acid before the addition of [14C]thymidine. 3. The inhibition of DNA synthesis by mycophenolic acid in L cells in vitro is reversed by guanine in a non-competitive manner, but not by hypoxanthine, xanthine or adenine. 4. The reversal of inhibition by guanine can be suppressed by hypoxanthine, 6-mercaptopurine and adenine. 5. Mycophenolic acid does not inhibit the incorporation of [14C]thymidine into DNA in suspensions of Landschütz and Yoshida ascites cells in vitro. 6. Mycophenolic acid inhibits the conversion of [14C]hypoxanthine into cold-acid-soluble and -insoluble guanine nucleotides in Landschütz and Yoshida ascites cells and also in L cells in vitro. There is some increase in the radioactivity of the adenine fraction in the presence of the antibiotic. 7. Mycophenolic acid inhibits the conversion of [14C]hypoxanthine into xanthine and guanine fractions in a cell-free system from Landschütz cells capable of converting hypoxanthine into IMP, XMP and GMP. 8. Preparations of IMP dehydrogenase from Landschütz ascites cells, calf thymus and LS cells are strongly inhibited by mycophenolic acid. The inhibition showed mixed type kinetics with Ki values of between 3·03×10−8 and 4·5×10−8m. 9. Evidence was also obtained for a partial, possibly indirect, inhibition by mycophenolic acid of an early stage of biosynthesis of purine nucleotides as indicated by a decrease in the accumulation of formylglycine amide ribonucleotide induced by the antibiotic azaserine in suspensions of Landschütz and Yoshida ascites cells and L cells in vitro.

Relationship between chromatin structure and replication in mouse L-cells

1980 ◽  
Vol 58 (12) ◽  
pp. 1359-1369 ◽  
Author(s):  
Rose Sheinin ◽  
G. Setterfield ◽  
I. Dardick ◽  
G. Kiss ◽  
M. Dubsky
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
De Novo ◽  
Nuclear Volume ◽  
Chromatin Protein ◽  
Structural Reorganization ◽  
L Cells ◽  
Inhibition Of Dna Synthesis ◽  
De Novo Protein Synthesis ◽  
Nuclear Enlargement

Mouse L-cells treated with cytosine arabinoside, hydroxyurea, fluorodeoxyuridine, methotrexate, or mitomycin C rapidly cease DNA synthesis and stop dividing. Such inhibition of DNA replication is followed by interruption of formation of lysine- and arginine-containing proteins, including chromatin-bound histones, and by a major reorganization of the heterochromatin of the central nucleoplasm, manifest as disaggregation of large clumps of this condensed chromatin. Morphometric analysis revealed both cell and nuclear enlargement in cells treated with such antimetabolites of DNA replication. These observations are in contrast to those made with WT-4 cells starved of isoleucine or treated with cycloheximide. Isoleucine depletion was associated with inhibition of DNA synthesis and continued increase of cell and nuclear volume, but not with massive disaggregation of heterochromatin. Cycloheximide produced inhibition of DNA synthesis and protoplasmic growth, and also prevented structural reorganization of chromatin. A model is presented which suggests that initiation of chromatin replication is associated with a process, dependent upon de novo protein synthesis, which results in chromatin disaggregation. This can be revealed by inhibition of the correct replication of chromatin DNA and chromatin protein.

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