Biochemical differences between staurosporine-induced apoptosis and premature mitosis.
Apoptosis is morphologically related to premature mitosis, an aberrant form of mitosis. Staurosporine, a potent protein kinase inhibitor, induces not only apoptotic cell death in a wide variety of mammalian cells but also premature initiation of mitosis in hamster cells that are arrested in S phase by DNA synthesis inhibitors. Here we report on the biochemical differences between the two phenomena commonly caused by staurosporine. Rat 3Y1 fibroblasts that had been arrested in S phase with hydroxyurea underwent apoptosis by treatment with staurosporine, whereas S-phase-arrested CHO cells initiated mitosis prematurely when similarly treated with a low concentration of staurosporine. Chromosome condensation occurred in both apoptosis (3Y1) and premature mitosis (CHO). However, neither formation of mitotic spindles nor mitosis-specific phosphorylation of MPM-2 antigens was observed in apoptosis of 3Y1 cells, unlike premature mitosis of CHO cells. The p34cdc2 kinase activated in normal and prematurely mitotic cells remained inactive in the apoptotic cells, probably because the active cyclin B/p34cdc2 complex was almost absent in the S-phase-arrested 3Y1 cells. The absence of intracellular activation of p34cdc2 in apoptosis was confirmed by immunohistochemical analyses using a specific antibody raised against Ser55-phosphorylated vimentin which is specifically phosphorylated by p34cdc2 during M phase. Furthermore, phosphorylation of histones H1 and H3, which is associated with mitotic chromosome condensation, did not occur in the apoptotic cells. These results indicate that the two phenomena, staurosporine-induced apoptosis and premature mitosis, are different in their requirement for p34cdc2 kinase activation and histone phosphorylation.
Yoshida
M
M
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.
Usui
T
T
Tsujimura
K
K
Inagaki
M
M
Beppu
T
T
Horinouchi
S
S
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D016466:CHO Cells / Q000187:drug effects / Q000201:enzymology; D002453:Cell Cycle / Q000187:drug effects; D002875:Chromosomes / Q000187:drug effects / Q000648:ultrastructure; D006224:Cricetinae; D003412:Cricetulus; D004261:DNA Replication / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D006657:Histones / Q000378:metabolism; D006918:Hydroxyurea / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects*; D010750:Phosphoproteins / Q000032:analysis; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D051381:Rats; D019311:Staurosporine / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D006657:Histones; D010750:Phosphoproteins; D016203:CDC2 Protein Kinase; D019311:Staurosporine; D006918:Hydroxyurea