Radiation and Environmental Biophysics

, Volume 33, Issue 1, pp 35–44 | Cite as

Damage at two levels of DNA folding measured by fluorescent halo technique in X-irradiated L5178Y-R and L5178Y-S cells. II. Repair

  • M. Kapiszewska
  • I. Szumiel
  • C. S. Lange


In the preceding paper we described the properties of nucleoids analyzed with the fluorescent halo assay at pH 6.9 and 9, as well as in the presence of reducing and chelating agents and after X-irradiation. We found analogies between the properties of type I and II nucleoids, as examined by Lebkowski and Laemmli (1982), and nucleoids analyzed with the fluorescent halo assay. We concluded that radiation-inflicted damage at two levels of DNA folding is measured at pH 6.9 and 9. In this paper we examined repair of damage to the nucleoid structure as assayed by the fluorescent halo method in X-irradiated L5178Y (LY) sublines; R (radiation resistant,D0=1.4 Gy) and S (radiation sensitive,D0=0.5 Gy). Halo diameters were measured after cell lysis in the presence of propidium iodide (PI; 0.5 to 50 µg/ml) at pH 6.9 and 9. The ability of DNA to be rewound at 10–50 µg/ml of PI was impaired by X-irradiation and partly restored during 90-min post-irradiation incubation, indicating damage to the superhelical structure and its partial restoration. The exponential time constants for repair were 10.1 min (LY-S, 6 Gy), 11.2 min (LY-R, 12 Gy), and 20.3 min (LY-s, 12 Gy) when measured at pH 9. In X-irradiated (12 Gy) LY-S cells, slower restoration of DNA supercoiling was observed at pH 9 than at pH 6.9. The presence of labile lesions at pH 9 did not prevent restoration of the higher-order DNA structure, as estimated from DNA rewinding at pH 6.9 in LY-S cells.


Iodide Propidium Iodide Environmental Physic Radiation Resistant Exponential Time 
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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • M. Kapiszewska
    • 1
  • I. Szumiel
    • 2
  • C. S. Lange
    • 3
  1. 1.Institute of Molecular BiologyJagiellonian UniversityKrakówPoland
  2. 2.Institute of Nuclear Chemistry and TechnologyWarszawaPoland
  3. 3.Department of Radiation OncologySUNY-Health Science Center at BrooklynBrooklynUSA

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