Evidence that Fas-induced apoptosis leads to S phase arrest.
Cell death by apoptosis is an efficient mechanism of eliminating unwanted or aberrant cells. Triggering of Fas, a member of the tumor necrosis factor (TNF) receptor superfamily, by anti-Fas antibodies or by the Fas ligand (FasL), has been shown to cause cell death by apoptosis. A recent study from our laboratory has demonstrated that Fas crosslinking leads to the dephosphorylation of the tumor suppressor retinoblastoma protein (Rb) and that this dephosphorylation is inhibited by calyculin A, a serine/threonine phosphatase inhibitor. In this investigation, we compared the effect of Fas crosslinking by CH11, an anti-Fas mAb, with two cyclin-dependent kinase (CDK) inhibitors, a peptide that specifically inhibits CDK2 (cdk2 inh) and roscovitine, which inhibits CDK2, CDC2, and CDK5. We illustrate that roscovitine induced DNA fragmentation, whereas cdk2 inh did not. In contrast to Fas-induced apoptosis, roscovitine-induced apoptosis was resistant to calyculin A. Both cdk2 inh and roscovitine induced cleavage of poly (ADP-ribose) polymerase (PARP) within 2 h. Roscovitine, however, led to the degradation of Rb, whereas cdk2 inh did not. Furthermore, both CH11 and roscovitine caused cell cycle arrest in S phase. In contrast, cdk2 inh did not have any effect on Jurkat cell cycle progression. Taken together, our results strongly suggest that the maintenance of Rb in its hyperphosphorylated form during S phase may be necessary for cell survival and that Rb dephosphorylation during S phase may constitute a crucial step in Fas-induced apoptosis.
N'cho
M
M
Department of Microbiology/Immunology, Indianapolis, IN 46202-5200, USA.
Brahmi
Z
Z
D017209:Apoptosis; D042846:CDC2-CDC28 Kinases; D002453:Cell Cycle; D002455:Cell Division; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors*; D053938:DNA Fragmentation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D008387:Marine Toxins; D010080:Oxazoles / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*; D011687:Purines / Q000494:pharmacology; D000077546:Roscovitine; D016196:S Phase; D019014:fas Receptor / Q000276:immunology / Q000378:metabolism*
D004791:Enzyme Inhibitors; D008387:Marine Toxins; D010080:Oxazoles; D011687:Purines; D019014:fas Receptor; D000077546:Roscovitine; C059041:calyculin A; D011065:Poly(ADP-ribose) Polymerases; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D010749:Phosphoprotein Phosphatases