In vitro heat exposure induces a redistribution of nuclear matrix proteins in human K562 erythroleukemia cells.
By using both conventional and confocal laser scanning microscopy with three monoclonal antibodies recognizing nuclear matrix proteins we have investigated by means of indirect fluorescence whether an incubation of isolated nuclei at the physiological temperature of 37 degrees C induces a redistribution of nuclear components in human K562 erythroleukemia cells. Upon incubation of isolated nuclei for 45 min at 37 degrees C, we have found that two of the antibodies, directed against proteins of the inner matrix network (M(r) 125 and 160 kDa), gave a fluorescent pattern different from that observed in permeabilized cells. By contrast, the fluorescent pattern did not change if nuclei were kept at 0 degrees C. The difference was more marked in case of the 160-kDa polypeptide. The fluorescent pattern detected by the third antibody, which recognizes the 180-kDa nucleolar isoform of DNA topoisomerase II, was unaffected by heat exposure of isolated nuclei. When isolated nuclear matrices prepared from heat-stabilized nuclei were stained by means of the same three antibodies, it was possible to see that the distribution of the 160-kDa matrix protein no longer corresponded to that observable in permeabilized cells, whereas the fluorescent pattern given by the antibody to the 125-kDa polypeptide resembled that detectable in permeabilized cells. The 180-kDa isoform of topoisomerase II was still present in the matrix nucleolar remnants. We conclude that a 37 degrees C incubation of isolated nuclei induces a redistribution of some nuclear matrix antigens and cannot prevent the rearrangement in the spatial organization of one of these antigens that takes place during matrix isolation in human erythroleukemia cells. The practical relevance of these findings is discussed.
Istituto di Anatomia Umana Normale, Università di Ferrara, Italy.
L M
LM
Neri
S
S
Santi
R A
RA
Marugg
B M
BM
Riederer
S
S
Capitani
A
A
Cataldi
A M
AM
Martelli
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D006358:Hot Temperature; D006801:Humans; D007211:Indoles; D004915:Leukemia, Erythroblastic, Acute; D008856:Microscopy, Fluorescence; D015530:Nuclear Matrix / Q000378:metabolism* / Q000648:ultrastructure; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000906:Antibodies; D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D007211:Indoles; D009687:Nuclear Proteins; C007293:DAPI