Locating of nucleic acid intercalators in yeast cells by image analysis combined fluorescence microscopy.
Intracellular distribution in the intact (not fixed) Saccharomyces cerevisiae cells of the nucleic acid intercalators (NAI) was studied using fluorescence microscopy combined with computer image analysis (ImageJ software, NIH, USA). Three NAI-the anthracycline anticancer drug doxorubicin (DR) along with the nucleic acid dyes ethidium (E) and 4',6-diamidino-2-phenylindole (DAPI)-were used. Staining pattern and ImageJ quantitative analysis data provided evidence that all three NAI were located in the nuclei and in the mitochondria. DR and E, in contrast to DAPI, may be bound to not only DNA, but to the mitochondrial membranes as well. Experiments on the combined application of DR+DAPI and E+DAPI have shown potential competition of DAPI with DR and E for binding sites in the nuclear and mitochondrial DNA. With the approach presented herein, the yeast cells of S. cerevisiae can be used as a model for locating intracellular sites of the fluorescing nucleic acid intercalators. This model may be of help in designing new DNA-targeted drugs and in preliminary studies of their interaction with eukaryotic cells.
G.K.Skryabin Institute of Biochemistry and Physiology of Microorganisms, RAS, Institutskaya Str., 5, Pushchino 142290, Russia. puchkov@ibpm.pushchino.ru
Evgeny O
EO
Puchkov
Millicent
M
McCarren
D001665:Binding Sites; D002467:Cell Nucleus; D003952:Diagnostic Imaging / Q000379:methods*; D004317:Doxorubicin; D004996:Ethidium; D007211:Indoles; D007364:Intercalating Agents; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria; D009696:Nucleic Acids / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012984:Software; D015003:Yeasts / Q000166:cytology / Q000235:genetics*
D007211:Indoles; D007364:Intercalating Agents; D009696:Nucleic Acids; C007293:DAPI; D004317:Doxorubicin; D004996:Ethidium