Abstract
The homeostasis of animals is regulated not only by the growth and differentiation of cells, but also by cell death through a process known as apoptosis. Apoptosis is mediated by members of the caspase family of proteases, and eventually causes the degradation of chromosomal DNA. A caspase-activated deoxyribonuclease (CAD) and its inhibitor (ICAD) have now been identified in the cytoplasmic fraction of mouse lymphoma cells. CAD is a protein of 343 amino acids which carries a nuclear-localization signal; ICAD exists in a long and a short form. Recombinant ICAD specifically inhibits CAD-induced degradation of nuclear DNA and its DNase activity. When CAD is expressed with ICAD in COS cells or in a cell-free system, CAD is produced as a complex with ICAD: treatment with caspase 3 releases the DNase activity which causes DNA fragmentation in nuclei. ICAD therefore seems to function as a chaperone for CAD during its synthesis, remaining complexed with CAD to inhibit its DNase activity; caspases activated by apoptotic stimuli then cleave ICAD, allowing CAD to enter the nucleus and degrade chromosomal DNA.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Jacobson, M. D., Weil, M. & Raff, M. C. Programmed cell death in animal development. Cell 88, 347–354 ( 1997).
Nagata, S. Apoptosis by death factor. Cell 88, 355– 365 (1997).
Wyllie, A. H., Kerr, J. F. R. & Currie, A. R. Cell death: the significance of apoptosis. Int. Rev. Cytol. 68, 251–306 (1980).
Compton, M. M. Abiochemical hallmark of apoptosis: internucleosomal degradation of the genome. Cancer Metast. Rev. 11, 105– 119 (1992).
Wyllie, A. H., Morris, R. G., Smith, A. L. & Dunlop, D. Chromatin cleavage in apoptosis: association with condensed chromatin morphology and dependence on macromolecular synthesis. J. Pathol. 142, 66–77 (1984).
Wyllie, A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284, 555– 556 (1980).
Peitsch, M. C. et al. Characterization of the endogenous deoxyribonuclease involved in nuclear DNA degradation during apoptosis (programmed cell death). EMBO J. 12, 371–377 ( 1993).
Montague, J. W., Hughes, F. J. & Cidlowski, J. A. Native recombinant cyclophilins A, B, and C degrade DNA independently of peptidylpropyl cis-trans-isomerase activity. Potential roles of cyclophilins in apoptosis. J. Biol. Chem. 272, 6677–66784 ( 1997).
Barry, M. & Eastman, A. Identification of deoxyribonuclease II as an endonuclease involved in apoptosis. Arch. Biochem. Biophys. 300, 440–450 ( 1993).
Nagata, S. & Golstein, P. The Fas death factor. Science 267, 1449–1456 ( 1995).
Itoh, N. et al. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis. Cell 66, 233– 243 (1991).
Schulze-Osthoff, K., Walczak, H., Dröge, W. & Krammer, P. H. Cell nucleus and DNA fragmentation are not required for apoptosis. J. Cell. Biol. 127, 15– 20 (1994).
Enari, M., Hug, H. & Nagata, S. Involvement of an ICE-like protease in Fas-mediated apoptosis. Nature 375, 78–81 ( 1995).
Enari, M., Talanian, R. V., Wong, W. W. & Nagata, S. Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis. Nature 380, 723– 726 (1996).
Longthorne, V. & Williams, G. Caspase activity is required for commitment to Fas-mediated apoptosis. EMBO J. 16, 3805–3812 (1997).
Armstrong, R. C. et al. Fas-induced activation of the cell death-related protease CPP32 is inhibited by Bcl-2 and by ICE family protease inhibitors. J. Biol. Chem. 271, 16850–16855 (1996).
Muzio, M. et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex. Cell 85, 817–827 ( 1996).
Boldin, M. P., Goncharov, T. M., Goltsev, Y. V. & Wallach, D. Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. Cell 85, 803–815 (1996).
Henkart, P. A. ICE family protease: mediators of all apoptotic cell death? Immunity 4, 195–201 ( 1996).
Fraser, A. & Evan, G. Alicense to kill. Cell 85, 781–784 (1996).
Martin, S. & Green, D. Protease activation during apoptosis: death by a thousand cuts. Cell 82, 349– 352 (1995).
Enari, M., Hase, A. & Nagata, S. Apoptosis by a cytosolic extract from Fas-activated cells. EMBO J. 14, 5201–5208 (1995).
Martin, S. J. et al. Cell-free reconstitution of Fas-, UV radiation- and ceramide-induced apoptosis. EMBO J. 14, 5191– 5200 (1995).
Liu, X., Zou, H., Slaughter, C. & Wang, X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89, 175– 184 (1997).
Sakahira, H., Enari, M. & Nagata, S. Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391, 96– 99 (1998).
Blanar, M. A. & Rutter, W. J. Interaction cloning: identification of a helix-loop-helix zipper protein that interacts with c-Fos. Science 256, 1014–1018 ( 1992).
Zakut, R. et al. Nucleotide sequence of the rat skeletal muscle actin gene. Nature 298, 857–859 ( 1982).
Dingwall, C. & Laskey, R. Nuclear targeting sequences—a consensus? Trends Biol. Sci. 16, 478– 481 (1991).
Cohen, J. J., Duke, R. C., Fadok, V. A. & Sellins, K. S. Apoptosis and programmed cell death in immunity. Annu. Rev. Immunol. 10, 267–293 ( 1992).
Shiokawa, D., Iwamatsu, A. & Tamura, S. Purification, characterization, and amino acid sequencing of DNase γ from rat spleen. Arch. Biochem. Biophys. 346, 15–20 (1997).
Batistatou, A. & Green, L. Internucleosomal DNA cleavage and neuronal cell survival/death. J. Cell Biol. 122, 523–532 (1993).
Mogil, R. et al. Role of DNA fragmentation in T cell activation-induced apoptosis in vitro and in vivo. J. Immunol. 152, 1674–1683 (1994).
Verma, I., Stevenson, J., Schwarz, E., Van Antwerp, D. & Miyamoto, S. ReI/NF-κB/IκB family: intimate tales of association and dissociation. Genes Dev. 9, 2723–2735 (1995).
Baldwin, A. The NF-κB and IκB proteins: new discoveries and insights. Annu. Rev. Immunol. 14, 649–681 (1996).
Beg, A. et al. IκB interacts with the nuclear localization sequences of the subunits of NF-κB: a mechanism for cytoplasmic retention. Genes Dev. 6, 1899–1913 (1992).
Wallis, R. et al. In vivo and in vitro characterization of overproduced colicin E9 immunity protein. Eur. J. Biochem. 207, 687–695 (1992).
Hartl, F.-U., Hlodan, R. & Langer, T. Molecular chaperones in protein folding: the art of avoiding sticky situations. Trends Biol. Sci. 19, 20–25 (1994).
Shi, G. et al. β-Subunits promote K+ channel surface expression through effects early in biosynthesis. Neuron 16, 843– 852 (1996).
Chen, P. & Hochstrasserr, M. Autocatalytic subunit processing couples active site formation in the 20S proteasome to completion of assembly. Cell 86, 961–972 (1996).
Ogasawara, J. et al. Lethal effect of the anti-Fas antibody in mice. Nature 364, 806–809 ( 1993).
Iwamatsu, A. S-carboxymethylation of proteins transferred onto polyvinylidine difluoride membranes followed by in situ protease digestion and amino acid microsequencing. Electrophoresis 13, 142– 147 (1992).
Iwamatsu, A. & Yoshida-Kuboomura, N. Systematic peptide fragmentation of polyvinylidine difluoride (PVDF)-immobilized proteins prior to microsequencing. J. Biochem. (Tokyo) 120, 29– 34 (1996).
Suda, T., Takahashi, T., Golstein, P. & Nagata, S. Molecular cloning and expression of the Fas ligand: a novel member of the tumor necrosis factor family. Cell 75, 1169 –1178 (1993).
Hager, D. A. & Burgess, R. R. Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Escherichai coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Analyt. Biochem. 109, 76– 86 (1980).
Acknowledgements
We thank R. V. Talanian for the caspase 3 expression system, M. A. Blanar for pGEX-2T[128/129], and S. Kumagai for secretarial assistance. This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture in Japan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Enari, M., Sakahira, H., Yokoyama, H. et al. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391, 43–50 (1998). https://doi.org/10.1038/34112
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/34112
This article is cited by
-
Novel meriolin derivatives activate the mitochondrial apoptosis pathway in the presence of antiapoptotic Bcl-2
Cell Death Discovery (2024)
-
An intracellular, non-oxidative factor activates in vitro chromatin fragmentation in pig sperm
Biological Research (2023)
-
Drug-tolerant persister cells in cancer: the cutting edges and future directions
Nature Reviews Clinical Oncology (2023)
-
Cold Stress Induces Apoptosis in Silver Pomfret via DUSP-JNK Pathway
Marine Biotechnology (2023)
-
Chronic exposure to methadone induces activated microglia and astrocyte and cell death in the cerebellum of adult male rats
Metabolic Brain Disease (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.