Localisation of DNA topoisomerase IIalpha in mouse erythroleukemia cells.
The presence of DNA topoisomerase IIalpha was investigated in interphase and metaphase mouse erythroleukemia (MEL) Friend-S cells, and in extracted with 25 mM lithium diiodosalicylate buffer (Lis) nuclei using indirect immunofluorescence. The results showed that DNA topoisomerase IIalpha is localised in the nuclei. In the metaphase cells, we found high concentrations of this enzyme in the mitotic chromosomes. Our results support the idea of the accumulation of DNA topoisomerase IIalpha at the end of the cell cycle. The extractions of nuclei with 25 mM Lis led to the complete depletion of DNA topoisomerase IIalpha from the residual nuclear matrix. Using a high dilution of the first antibody, we established that the high level of heterochromatin compactisation in the interphase nuclei is caused by the high concentration of DNA topoisomerase IIalpha.
Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
E C
EC
Ivanova
R M
RM
Donev
L P
LP
Djondjurov
D000818:Animals; D000951:Antigens, Neoplasm; D002467:Cell Nucleus / Q000201:enzymology; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004268:DNA-Binding Proteins; D019084:Fluorescent Antibody Technique, Indirect; D005622:Friend murine leukemia virus; D006570:Heterochromatin / Q000201:enzymology; D007399:Interphase; D007527:Isoenzymes / Q000378:metabolism*; D004915:Leukemia, Erythroblastic, Acute / Q000201:enzymology* / Q000473:pathology; D008677:Metaphase; D051379:Mice; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D006570:Heterochromatin; D007527:Isoenzymes; D004250:DNA Topoisomerases, Type II