LOCALIZATION OF DNA TOPOISOMERASE-II IN CHINESE-HAMSTER FIBROBLASTS BY CONFOCAL AND ELECTRON-MICROSCOPY

The localization of the 170- and the 180-kDa isoforms of the enzyme DNA topoisomerase II in growing Chinese hamster fibroblasts has been studied by confocal immunofluorescence microscopy and immunogold electron microscopy after labeling with affinity-purified isoform-specific polyclonal antibodies. Immunofluorescence and immunogold studies, together with quantitative image analysis, show that the two isoforms are present in the nucleoplasm and in the nucleolus. In the nucleoplasm both isoforms are frequently localized at the periphery of heterochromatin regions. In the nucleolus the Immunofluorescence and immunogold signals relative to surface area are higher than in the nucleoplasm; both isoforms are localized predominantly in the fibrillar zones. During mitosis both isoforms remain detectable in the cytoplasm. The differential expression of the two isoforms during the cell cycle, observed in other studies, suggests that they have different functions, and their presence in both the nucleoplasm and the nucleolus suggests that these functions are required in both of these nuclear Compartments. © 1903 Academic Press, Inc.