Wiseman, Andrew and Attardi, Giuseppe (1978) Reversible tenfod reduction in mitochondrial DNA content of human cells treated with ethidium bromide. Molecular and General Genetics MGG, 167 (1). pp. 51-63. ISSN 0026-8925. doi:10.1007/bf00270321. https://resolver.caltech.edu/CaltechAUTHORS:20221004-680171300.15
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Abstract
Cells of the human line VA₂-B in suspension culture have been treated with very low concentrations of ethidium bromide for the purpose of reducing the amount of mitochondrial DNA (mit-DNA) per cell. Cells maintained in the presence of 5 ng/ml ethidium bromide grew at a normal rate for three days; thereafter, their doubling time gradually increased to a stable value of about 60 h. In these cells, the rate of ³H thymidine incorporation into mit-DNA decreased very rapidly to ∼60% of the normal, and remained thereafter at this level, while the amount of mit-DNA per cell stabilized around a level of 70–80% of the control. In cells long-term treated with 5 ng/ml ethidium bromide, the rate of mitochondrial protein synthesis was about 35% of the normal, and the cytochrome c oxidase activity about 50% of the control. Cells treated with 20 ng/ml of the drug underwent 3–4 cell doublings at control rates, then gradually stopped growing, and eventually died. In these cells, the rate of incorporation of ³H thymidine into mit-DNA was reduced to 50% of the control value after 10 min treatment with ethidium bromide, and became barely detectable after three cell doublings. At this time, the cells had on the average less than 10% of the control amount of mit-DNA, the rate of mitochondrial protein synthesis was reduced to 3% of the normal, and the specific activities of cytochrome c oxidase and rutamycin-sensitive ATPase were less than 20% of the control values. In spite of these marked changes, the cells exhibited only a 20–30% loss in cell viability, as estimated by cloning efficiency, after three days of exposure to the drug. Cells treated with ethidium bromide at 20 ng/ml for three days, and then transferred to drug-free medium, recovered a near-to-normal growth rate and cloning efficiency and a near-to-normal rate of synthesis and amount of mit-DNA in about five days.
Item Type: | Article | ||||||
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Issue or Number: | 1 | ||||||
DOI: | 10.1007/bf00270321 | ||||||
Record Number: | CaltechAUTHORS:20221004-680171300.15 | ||||||
Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20221004-680171300.15 | ||||||
Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | ||||||
ID Code: | 117241 | ||||||
Collection: | CaltechAUTHORS | ||||||
Deposited By: | Tony Diaz | ||||||
Deposited On: | 12 Oct 2022 23:01 | ||||||
Last Modified: | 12 Oct 2022 23:01 |
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