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Metal ions protect DNA against strand breakage induced by fast neutrons.

Single and double strand breaks (SSB and DSB) are induced by fast neutrons in plasmid (pBR322) DNA in 1 mM potassium phosphate buffer (pH 7.25). Increasing the concentration of monovalent (Na+, Cs+, Li+), divalent (Mg2+, Ca2+) and trivalent (Al3+, Co3+ (NH3)6) metal cations strongly decreases the yield of DSB. The extent of the observed protection depends on the valence of the cation. The production of SSB is only slightly decreased, except for Al3+ and Co3+ (NH3)6, whose effects are particularly large (complete protection at 1 and 0.1 mM respectively). Circular dichroism spectra show that Al3+ induces an important structural change of DNA at the ion concentration where the protection becomes total. This change is probably a condensation (collapse), as in the well-known case of Co3+ (NH3)6. Our results suggest two mechanisms of protection by metal ions: (i) the induction of structural changes of DNA, that render less accessible the critical sites of attack by OH. radicals; and (ii) the stabilization of the double helical regions between two close-set nicks on opposite strands, that hinders the effective double strand breakage of DNA.

Cations↗

Marked effect of buffers on yield of single- and double-strand breaks in DNA irradiated at room temperature and at 77 K.

We have shown that the protocol for handling plasmid (pBR 322) DNA for radiation studies, which normally involves using quite high concentrations of Tris or phosphate buffers, is equally satisfactory in the absence of added buffers provided samples are stored at 0 degree C, or used directly. On exposure of aqueous solutions at 77 K, Tris buffer acts as a weak protecting agent, but phosphate buffers at the same pH act as remarkably effective sensitizing agents, giving ca. 100-fold increases in strand breaks. At room temperature there is again a marked difference between the aqueous and buffered systems, but in this case both the Tris and the phosphate systems are protective. Possible reasons for these contrasting results are discussed, and the advantages of using simple aqueous solutions are stressed.

Buffers↗

Incorporation of microdosimetric concepts into a biologically-based model of radiation carcinogenesis.

The generalised state-vector model of radiation carcinogenesis (SVM) simulates radiation induced biological effects by expressing the transition rates between the various initiation and promotion stages in terms of dose rate for low and high linear energy transfer (LET) particles. In the present work, the SVM has been reformulated to incorporate single track characteristics of particles with varying LET. Transition rates of the initiation phase were expressed as functions of LET by describing the complexity and clustering of DNA double strand breaks (DSBs) and its effect on repair kinetics, while the promotion phase was reformulated based on a multi-target single-hit hypothesis. Such an approach allows the consideration of hit frequencies and the variability of the specific energy and LET spectra of radon progeny alpha particles in bronchial target cells for different exposure conditions.

Alpha Particles↗

Selective stimulation of one of the mechanisms for genetic recombination of bacteriophage S13.

In certain recombination-deficient (Rec(-)) bacterial strains genetic recombination of bacteriophage S13 is reduced, but the existence of some residual recombination has suggested that there is a secondary mechanism of phage recombination that is still functioning. In these Rec(-)strains it is found that there is no stimulation of recombination by irradiation of the parental phage with ultraviolet light, in contrast to the large increase found when irradiated phage particles infect a Rec(+) host. This selective stimulation of phage recombination in the Rec(+) but not in the Rec(-) strains supports the view that the phage uses at least two mechanisms of genetic recombination.

Coliphages↗

Multiplicity reactivation as a test for recombination function.

Multiplicity reactivation of bacteriophage inactivated by ultraviolet light is dependent on the recombination function of either the host bacterial cell or the infecting bacteriophage. Absence of both recombination systems leads to a loss of multiplicity reactivation.

Coliphages↗

Replication of the bacterial chromosome: location of new initiation sites after irradiation.

New loci of replication along the bacterial chromosome are observed after irradiation of Escherichia coli. It was conjectured that, after X-irradiation, the new initiation site was random with respect to the fixed-origin, whereas, after ultraviolet light exposure, it was selective and appeared to be from the fixed-origin. Evidence presented here shows that, after X-irradiation of E. coli, the new initiation site(s) for the onset of deoxyribonucleic acid replication is induced at chromosomal regions not restricted to the fixed-origin. After ultraviolet light exposure, the new initiation site is preferentially from the fixed-origin. In these studies amino acid starvation was used to synchronize chromosome replication and to allow for differential radioisotopic labeling of the chromosomal origin and terminus. To facilitate interpretation, growing cells actively replicating their chromosome were compared with cells lacking growth points at the time of irradiation. The role of these new replication sites in the observed kinetics of deoxyribonucleic acid replication following X-ray or ultraviolet light exposure is discussed.

Carbon Isotopes↗