The effect of sodium tetrathionate stabilization on the distribution of three nuclear matrix proteins in human K562 erythroleukemia cells.
Using both conventional fluorescence and confocal laser scanning microscopy we have investigated whether or not stabilization of isolated human erythroleukemic nuclei with sodium tetrathionate can maintain in the nuclear matrix the same spatial distribution of three polypeptides (M(r) 160 kDa and 125 kDa, previously shown to be components of the internal nuclear matrix plus the 180-kDa nucleolar isoform of DNA topoisomerase II) as seen in permeabilized cells. The incubation of isolated nuclei in the presence of 2 mM sodium tetrathionate was performed at 0 degrees C or 37 degrees C. The matrix fraction retained 20-40% of nuclear protein, depending on the temperature at which the chemical stabilization was executed. Western blot analysis revealed that the proteins studied were completely retained in the high-salt resistant matrix. Indirect immunofluorescence experiments showed that the distribution of the three antigens in the final matrix closely resembled that detected in permeabilized cells, particularly when the stabilization was performed at 37 degrees C. This conclusion was also strengthened by analysis of cells, isolated nuclei and the nuclear matrix by means of confocal laser scanning microscopy. We conclude that sodium tetrathionate stabilization of isolated nuclei does not alter the spatial distribution of some nuclear matrix proteins.
Neri
L M
LM
Istituto di Anatomia Umana Normale, Università di Ferrara, Italy.
Riederer
B M
BM
Marugg
R A
RA
Capitani
S
S
Martelli
A M
AM
D015153:Blotting, Western; D002463:Cell Membrane Permeability; D004273:DNA, Neoplasm / Q000096:biosynthesis; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000378:metabolism*; D015530:Nuclear Matrix / Q000378:metabolism*; D013776:Tetrathionic Acid / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D007202:Indicators and Reagents; D009363:Neoplasm Proteins; D013776:Tetrathionic Acid