[Changes in the induction and repair of double-stranded DNA breaks in pro- and eukaryote cells. I. Use of a Zimm elastoviscosimeter to study induction of double-stranded DNA breaks in gamma-irradiated Escherichia coli cells].
The technique of elastoviscosimetry allows to study the induction of double-strand breaks in DNA of E. coli at low doses (on the order of D37). The dose dependence of retardation time to shows a characteristic maximum. It is shown that the ascending part of the curve is due to the phenomenon of relaxation of supercoiled DNA in the bacterial nucleoid. Relaxation is effected by different gamma-induced damages in DNA which are not double-strand breaks. The position of the maximum yields the average dose for the formation of the first double-strand break, which transforms the circular DNA into a linear chain. The descending part of the dose curve is explained by accumulation of additional double-strand breaks. The gamma-irradiation and lysis of cells was performed in different media. It was found that only in the case when the action of nucleases was substantially (but not completely) inhibited, the position of the maximum of the dose dependent of retardation time coincides satisfactorily with the value of D37 (14.5 +/- 2.3 and 12.5 +/- 3 krad correspondingly). If the medium does not contain inhibitors of nucleases then the position of the maximum corresponds to a 4.2 times lower dose of gamma-rays. This shows that double-strand breaks in gamma-irradiated DNA are generated mainly by enzymes participating in repair processes and that the first double-strand breaks seems to be the true reason of lethality because of inability to be repaired.
D004260:DNA Repair; D004269:DNA, Bacterial / Q000528:radiation effects*; D004278:DNA, Superhelical / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D004548:Elasticity; D004720:Endonucleases / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000528:radiation effects*; D005092:Exonucleases / Q000378:metabolism; D005720:Gamma Rays; D008722:Methods; D014783:Viscosity
D004269:DNA, Bacterial; D004278:DNA, Superhelical; D004720:Endonucleases; D005092:Exonucleases
Izuchenie induktsii i reparatsii dvunitevykh razryvov DNK v kletkakh pro- i éukariot. I. Primenenie élastoviskozimetra Zimma dlia izucheniia induktsii dvunitevykh razryvov DNK v gamma-obluchennykh kletkakh Escherichia coli.