Expression of the 170-kDa and 180-kDa isoforms of DNA topoisomerase II in resting and proliferating human lymphocytes.
The expression of the 170-kDa (alpha) and the 180-kDa (beta) isoforms of DNA topoisomerase II (topo II) was investigated with two specific monoclonal antibodies (MoAbs) in human peripheral blood lymphocytes (PBL), before and after phytohaemoagglutinin (PHA) stimulation. Binding of each MoAb was detected by indirect immunofluorescence labelling and quantified with flow cytometry. In resting PBL, the intensity of immunostaining was very low for both isozymes; however, topo IIbeta-associated immunofluorescence was about 2.5 times significantly higher (P<0.001) than that associated with the alpha isoform. Between 48 and 72 h of PHA stimulation, when the highest percentage of cells in S and G2+M phases was found, the levels of topo IIalpha and beta increased up to about 30 and 10 times the value measured in resting PBL, respectively. Thus, the two isoforms reached comparable immunofluorescence values. At longer stimulation periods (96-120 h), topo IIalpha immunofluorescence was not significantly changed, while that relative to topo IIbeta declined to about 50% of the peak value (P<0.02). At this time however, topo IIalpha-associated immunofluorescence was not significantly different from that related to the beta isozyme. These results suggest that in resting PBL topo IIalpha is required at levels lower than topo IIbeta, while in proliferating lymphocytes both isoforms are expressed to significantly higher levels.
Prosperi
E
E
Centro di Studio per l'Istochimica del C.N.R., Pavia, Italy.
Negri
C
C
Marchese
G
G
Ricotti
G C
GC
D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008214:Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000494:pharmacology; D020033:Protein Isoforms / Q000378:metabolism
D010835:Phytohemagglutinins; D020033:Protein Isoforms; D004250:DNA Topoisomerases, Type II