Characteristics of the enzymatic breakdown of DNA in Escherichia coli in response to ionizing radiation.
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Biomedical subjects
Publications and source records attributed to E C Pollard.
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Cells of Escherichia coli have been successfully banded in CsCl density gradients and a portion of the population reclaimed in a viable state. Differentiation between two strains of this organism in a CsCl density gradient has been demonstrated also. Several studies were undertaken to see whether differences could be detected between two samples of cells of the same strain which had been subjected to different conditions. The results were as follows: (a) Introduction of a heavy label (5-bromouracil) into the DNA during a 90 minute period did not produce an observable change in cell density. (b) Removal of a required amino acid from the growth medium of an E. coli auxotroph resulted in an increase in both the density and heterogeneity of the cells. (c) Exposure of cells to 27 kr of gamma radiation, followed by a period during which portions of both DNA and RNA were lost, yielded two distinct bands, one at the normal position in the gradient and the other shifted to a lighter region.
DNA degradation and its temperature dependence as a function of linear energy transfer were studied in Excherichia coli using fast proton irradiation as the initiating agent. The data indicate that radiation-induced DNA degradation can proceed by two processes. The first, or fast component, begins immediately after irradiation with (60)Co gamma-rays or with fast protons at doses less than 10(10) protons/cm(2). The rate is high and involves a maximum of about 50% degradation. It is elicited more efficiently by protons of high linear energy transfer. The second, or slow component, results from higher doses of fast proton bombardment. There is a delay between irradiation and the initiation of this slower component, but 100% of the DNA complement is degraded. The data indicate that both processes are enzyme-mediated, the first probably by normal DNA-related activity and the second by DNAase activity.
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