Preferential Accumulation of Cytoplasmic Protein Species during Inhibition of Deoxyribonucleic Acid Synthesis in Temperature-Sensitive Chinese Hamster Cells in Culture

1974 ◽  
Vol 2 (5) ◽  
pp. 864-864
Author(s):  
MANUEL RLEBER ◽  
JOSEFINA BACALAO ◽  
BASIL J. SMITH
Keyword(s):  
Deoxyribonucleic Acid ◽  
Chinese Hamster ◽  
Acid Synthesis ◽  
Temperature Sensitive ◽  
Cytoplasmic Protein ◽  
Protein Species ◽  
Chinese Hamster Cells ◽  
Cells In Culture ◽  
Preferential Accumulation ◽  
Deoxyribonucleic Acid Synthesis

Induction of cellular deoxyribonucleic acid synthesis in butyrate-treated cells by simian virus 40 deoxyribonucleic acid

1981 ◽  
Vol 1 (11) ◽  
pp. 1038-1047
Author(s):  
S Kawasaki ◽  
L Diamond ◽  
R Baserga
Keyword(s):  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Simian Virus 40 ◽  
Acid Synthesis ◽  
Simian Virus ◽  
S Phase ◽  
Temperature Sensitive ◽  
3T3 Cells ◽  
Deoxyribonucleic Acid Synthesis ◽  
G1 Cells

Sodium butyrate (3 mM) inhibited the entry into the S phase of quiescent 3T3 cells stimulated by serum, but had no effect on the accumulation of cellular ribonucleic acid. Simian virus 40 infection or manual microinjection of cloned fragments from the simian virus 40 A gene caused quiescent 3T3 cells to enter the S phase even in the presence of butyrate. NGI cells, a line of 3T3 cells transformed by simian virus 40, grew vigorously in 3 mM butyrate. Homokaryons were formed between G1 and S-phase 3T3 cells, Butyrate inhibited the induction of deoxyribonucleic acid synthesis that usually occurs in B1 nuclei when G1 cells are fused with S-phase cells. However, when G1 3T3 cells were fused with exponentially growing NGI cells, the 3T3 nuclei were induced to enter deoxyribonucleic acid synthesis. In tsAF8 cells, a ribonucleic acid polymerase II mutant that stops in the G1 phase of the cell cycle, no temporal sequence was demonstrated between the butyrate block and the temperature-sensitive block. These results confirm previous reports that certain virally coded proteins can induce cell deoxyribonucleic acid synthesis in the absence of cellular functions that are required by serum-stimulated cells. Our interpretation of these data is that butyrate inhibited cell growth by inhibiting the expression of genes required for the G0 leads to G1 leads to S transition and that the product of the simian virus 40 A gene overrode this inhibition by providing all of the necessary functions for the entry into the S phase.

scholarly journals Flow cytometric analysis of bromodeoxyuridine-substituted cells stained with 33258 Hoechst.

1977 ◽  
Vol 25 (7) ◽  
pp. 927-934 ◽  
Author(s):  
S A Latt ◽  
Y S George ◽  
J W Gray
Keyword(s):  
Flow Cytometry ◽  
Cell Growth ◽  
Deoxyribonucleic Acid ◽  
Flow Cytometric Analysis ◽  
Chinese Hamster ◽  
Acid Synthesis ◽  
Whole Cells ◽  
Flow Cytometric ◽  
Deoxyribonucleic Acid Synthesis ◽  
Average Fluorescence

This paper describes a flow-cytometric application of the quenching of fluorescence from 33258 Hoechst stained Chinese hamster ovary-line cells due to the incorporation of 5-bromo-deoxyuridine (BrdU) into the cellular deoxyribonucleic acid. Cells were grown for 24 hr in medium containing BrdU in concentrations ranging from 1 x 10(-8) to 1 x 10(-4) M. For each concentration we measured the average fluorescence as determined by flow cytometry, the extent of BrdU substitution and the effect of the BrdU on cell growth. We determined that a BrdU concentration of 1 x 10(-5) M resulted in sufficient substitution to quench the fluorescence from 33258 Hoechst by a factor of 4, allowing discrimination between cycling and noncycling cells. The extent of BrdU substitution after growth for 24 hr in this concentration of BrdU was 64%. These data indicate the feasibility of detecting deoxyribonucleic acid synthesis in whole cells using the 33258 Hoechst-BrdU methodology.

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