Radiolysis of chromatin extracted from cultured mammalian cells: production of alkali-labile strand damage in DNA.
Chromatin has been isolated from cultured Chinese-hamster lung fibroblasts as an expanded aqueous gel. The DNA in isolated chromatin has been examined by sedimentation on alkaline sucrose gradients. The average molecular weight of the DNA has been determined to be 50 million. gamma-irradiation of isolated chromatin degrades the DNA to lower molecular weight. The yield of single-strand breaks in the DNA is 0.02 single-strand breaks per krad-10(6) dalton, calculated from a dose-range of &--400 krad and covering a DNA molecular weight range of 2 X 10(7)-1.4 X 10(5). There is a considerable difference in the efficiency of the formation of single-strand breaks in DNA irradiated as isolated chromatin compared with chromatin irradiated in whole cells before isolation. For isolated chromatin, values of 6 dV per break have been calculated compared with about 80 eV per break for chromatin irradiated in whole cells, which suggest a large contribution from indirect action by aqueous radicals in isolated chromatin.
D000818:Animals; D002460:Cell Line; D002588:Cesium Radioisotopes; D002843:Chromatin / Q000528:radiation effects*; D003037:Cobalt Radioisotopes; D004277:DNA, Single-Stranded / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D005720:Gamma Rays; D066298:In Vitro Techniques
D002588:Cesium Radioisotopes; D002843:Chromatin; D003037:Cobalt Radioisotopes; D004277:DNA, Single-Stranded