28S ribosome degradation in lymphoid cell apoptosis: evidence for caspase and Bcl-2-dependent and -independent pathways.
Apoptosis, a physiological form of cell death, is characterized by the activation of a program that kills cells and recycles their constituents. We have used thymoma cell lines to examine the role of Bcl-2 and caspases in ribosomal destruction during apoptosis. Glucocorticoid- and calcium ionophore (A23187)-induced apoptosis of S49 Neo cells resulted in both 28S rRNA and DNA degradation. Interestingly, anisomycin, a potent protein synthesis inhibitor, also induced 28S rRNA and DNA fragmentation suggesting that the responsible nucleases are present in the viable cells and become activated during apoptosis. The anti-apoptotic protein, Bcl-2, inhibited both glucocorticoid- and anisomycin-induced DNA and 28S rRNA degradation but could not protect against A23187-induced nucleic acid degradation. We next examined the role of caspase activation in the generation of 28S rRNA degradation through the use of ZVAD, a general caspase inhibitor. Under conditions where ZVAD substantially decreased 28S rRNA degradation induced by glucocorticoid or anisomycin, no decrease was observed when A23187 was used to induce apoptosis. Surprisingly, RNA degradation, like DNA degradation, occurs exclusively in shrunken lymphocytes but not those with normal cell volume despite equivalent exposure of the cells to the apoptotic signals. Together, these findings indicate the ribosome is a specific target for death effectors during apoptosis and that a caspase/Bcl-2-independent pathway exists to activate its destruction.
Laboratory of Signal Transduction, National Institute of Environmental Health Sciences, National Institute of Health, MD E2-02, Research Triangle Park, NC 27709, USA.
K L
KL
King
C M
CM
Jewell
C D
CD
Bortner
J A
JA
Cidlowski
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D000841:Anisomycin / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology*; D015152:Blotting, Northern; D020169:Caspases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004273:DNA, Neoplasm / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D012339:RNA, Ribosomal, 28S / Q000378:metabolism*; D013945:Thymoma; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000201:enzymology
D000590:Amino Acid Chloromethyl Ketones; D015853:Cysteine Proteinase Inhibitors; D004273:DNA, Neoplasm; D011500:Protein Synthesis Inhibitors; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D012339:RNA, Ribosomal, 28S; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D000841:Anisomycin; D020169:Caspases