© The Rockefeller University Press, 0022-1007/1997/10/1107/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 7, October 6, 1997 1107-1117
Activation of CPP32-like Proteases Is Not Sufficient to Trigger Apoptosis: Inhibition of Apoptosis by Agents that Suppress Activation of AP24, but Not CPP32-like Activity
Susan C. Wright*,
Ute Schellenberger*,
Hong Wang*,
David H. Kinder
,
Jamil W. Talhouk*, and
James W. Larrick*
From the * Palo Alto Institute of Molecular Medicine, Mountain View, California 94043; and College of Pharmacy, Ohio Northern University, Ada, Ohio 45810
The 24-kD apoptotic protease (AP24) is a serine protease that is activated during apoptosis and has the capacity to activate internucleosomal DNA fragmentation in isolated nuclei. This study examined the following: (a) the functional relationship between AP24 and the CPP32-like proteases of the caspase family; and (b) whether activation of CPP32-like proteases is sufficient to commit irreversibly a cell to apoptotic death. In three different leukemia cell lines, we showed that agents that directly (carbobenzoxy-Ala-Ala-borophe (DK120) or indirectly inhibit activation of AP24 (protein kinase inhibitors, basic fibroblast growth factor, tosylphenylalaninechloromethylketone, and caspase inhibitors) protected cells from apoptosis induced by TNF or UV light. Only the caspase inhibitors, however, prevented activation of CPP32-like activity as revealed by cleavage of the synthetic substrate, DEVD-pNa, by cell cytosols, and also by in vivo cleavage of poly (ADP-ribosyl) polymerase, a known substrate of CPP32. Activation of DEVD-pNa cleaving activity without apoptosis was also demonstrated in two variants derived from the U937 monocytic leukemia in the absence of exogenous inhibitors. Cell-permeable peptide inhibitors selective for CPP32-like proteases suppressed AP24 activation and apoptotic death. These findings indicate that CPP32-like activity is one of several upstream signals required for AP24 activation. Furthermore, activation of CPP32-like proteases alone is not sufficient to commit irreversibly a cell to apoptotic death under conditions where activation of AP24 is inhibited.
Address correspondence to Dr. S.C. Wright, Palo Alto Institute of Molecular Medicine, 2462 Wayandotle Street, Mountain View, CA 94043.
1 Abbreviations used in this paper: ADPRT, ADP ribosyl transferase; CaM-KII, calcium/calmodulin-dependent protein kinase II; cpDEVD-CHO, cell-permeable Asp-Glu-Val-Asp-aldehyde; DEVD(meth)-cmk, side chain fully methylated carbobenzoxy-Asp-Glu-Val-Asp-chloromethylketone; ICE, interleukin-1β converting enzyme; PARP, poly (ADP-ribosyl) polymerase; SMase, sphingomyelinase; TPCK, tosylphenylalaninechloromethylketone; VAD-cmk, acetyl-Val-Ala-Asp-chloromethylketone.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Facebook 
Reddit 
Technorati 
Twitter What's this?
This article has been cited by other articles:
-
Jurewicz, A., Matysiak, M., Tybor, K., Kilianek, L., Raine, C. S., Selmaj, K.
(2005). Tumour necrosis factor-induced death of adult human oligodendrocytes is mediated by apoptosis inducing factor. Brain
128: 2675-2688
[Abstract]
[Full Text]
-
Murn, J., Urleb, U., Mlinaric-Rascan, I.
(2004). Internucleosomal DNA cleavage in apoptotic WEHI 231 cells is mediated by a chymotrypsin-like protease. GENES CELLS
9: 1103-1111
[Abstract]
[Full Text]
-
Suzuki, E., Handa, K., Toledo, M. S., Hakomori, S.
(2004). Sphingosine-dependent apoptosis: A unified concept based on multiple mechanisms operating in concert. Proc. Natl. Acad. Sci. USA
101: 14788-14793
[Abstract]
[Full Text]
-
Yoshida, A., Urasaki, Y., Waltham, M., Bergman, A.-C., Pourquier, P., Rothwell, D. G., Inuzuka, M., Weinstein, J. N., Ueda, T., Appella, E., Hickson, I. D., Pommier, Y.
(2003). Human Apurinic/Apyrimidinic Endonuclease (Ape1) and Its N-terminal Truncated Form (AN34) Are Involved in DNA Fragmentation during Apoptosis. J. Biol. Chem.
278: 37768-37776
[Abstract]
[Full Text]
-
Mildner, M., Eckhart, L., Lengauer, B., Tschachler, E.
(2002). Hepatocyte Growth Factor/Scatter Factor Inhibits UVB-induced Apoptosis of Human Keratinocytes but Not of Keratinocyte-derived Cell Lines via the Phosphatidylinositol 3-Kinase/AKT Pathway. J. Biol. Chem.
277: 14146-14152
[Abstract]
[Full Text]
-
Foghsgaard, L., Wissing, D., Mauch, D., Lademann, U., Bastholm, L., Boes, M., Elling, F., Leist, M., Jaattela, M.
(2001). Cathepsin B Acts as a Dominant Execution Protease in Tumor Cell Apoptosis Induced by Tumor Necrosis Factor. JCB
153: 999-1010
[Abstract]
[Full Text]
-
Mathiasen, I. S., Lademann, U., Jaattela, M.
(1999). Apoptosis Induced by Vitamin D Compounds in Breast Cancer Cells Is Inhibited by Bcl-2 but Does Not Involve Known Caspases or p53. Cancer Res.
59: 4848-4856
[Abstract]
[Full Text]
-
Jun, C.-D., Oh, C.-D., Kwak, H.-J., Pae, H.-O., Yoo, J.-C., Choi, B.-M., Chun, J.-S., Park, R.-K., Chung, H.-T.
(1999). Overexpression of Protein Kinase C Isoforms Protects RAW 264.7 Macrophages from Nitric Oxide-Induced Apoptosis: Involvement of c-Jun N-Terminal Kinase/Stress-Activated Protein Kinase, p38 Kinase, and CPP-32 Protease Pathways. J. Immunol.
162: 3395-3401
[Abstract]
[Full Text]
-
Escargueil-Blanc, I., Andrieu-Abadie, N., Caspar-Bauguil, S., Brossmer, R., Levade, T., Negre-Salvayre, A., Salvayre, R.
(1998). Apoptosis and Activation of the Sphingomyelin-Ceramide Pathway Induced by Oxidized Low Density Lipoproteins Are Not Causally Related in ECV-304 Endothelial Cells. J. Biol. Chem.
273: 27389-27395
[Abstract]
[Full Text]
-
Tang, D., Gururajan, R., Kidd, V. J.
(1998). Phosphorylation of PITSLRE p110 Isoforms Accompanies Their Processing by Caspases during Fas-mediated Cell Death. J. Biol. Chem.
273: 16601-16607
[Abstract]
[Full Text]
-
Park, C.-S., Yashiro, Y., Tai, X.-G., Toyo-oka, K., Hamaoka, T., Yagita, H., Okumura, K., Neben, S., Fujiwara, H.
(1998). Differential Involvement of a Fas-CPP32-Like Protease Pathway in Apoptosis of TCR/CD9-Costimulated, Naive T Cells and TCR-Restimulated, Activated T Cells. J. Immunol.
160: 5790-5796
[Abstract]
[Full Text]
