Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death
Caspase-activated DNase (CAD) is a major apoptotic nuclease, responsible for DNA fragmentation and chromatin condensation during apoptosis. CAD is normally activated in apoptosis as a result of caspase cleavage of its inhibitory chaperone ICAD. Other aspects of CAD regulation are poorly understood....
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| Published in | The Journal of biological chemistry Vol. 289; no. 45; pp. 31617 - 31623 |
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| Main Authors | , , , , , , |
| Format | Journal Article |
| Language | English |
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01.11.2014
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| Abstract | Caspase-activated DNase (CAD) is a major apoptotic nuclease, responsible for DNA fragmentation and chromatin condensation during apoptosis. CAD is normally activated in apoptosis as a result of caspase cleavage of its inhibitory chaperone ICAD. Other aspects of CAD regulation are poorly understood. In particular, it has been unclear whether direct CAD activation in non-apoptotic living cells can trigger cell death. Taking advantage of the auxin-inducible degron (AID) system, we have developed a suicide system with which ICAD is rapidly degraded in living cells in response to the plant hormone auxin. Our studies demonstrate that rapid ICAD depletion is sufficient to activate CAD and induce cell death in DT40 and yeast cells. In the vertebrate cells, ectopic CAD activation triggered caspase activation and subsequent hallmarks of caspase-dependent apoptotic changes, including phosphatidylserine exposure and nuclear fragmentation. These observations not only suggest that CAD activation drives apoptosis through a positive feedback loop, but also identify a unique suicide system that can be used for controlling gene-modified organisms. |
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| AbstractList | Caspase-activated DNase (CAD) is a major apoptotic nuclease, responsible for DNA fragmentation and chromatin condensation during apoptosis. CAD is normally activated in apoptosis as a result of caspase cleavage of its inhibitory chaperone ICAD. Other aspects of CAD regulation are poorly understood. In particular, it has been unclear whether direct CAD activation in non-apoptotic living cells can trigger cell death. Taking advantage of the auxin-inducible degron (AID) system, we have developed a suicide system with which ICAD is rapidly degraded in living cells in response to the plant hormone auxin. Our studies demonstrate that rapid ICAD depletion is sufficient to activate CAD and induce cell death in DT40 and yeast cells. In the vertebrate cells, ectopic CAD activation triggered caspase activation and subsequent hallmarks of caspase-dependent apoptotic changes, including phosphatidylserine exposure and nuclear fragmentation. These observations not only suggest that CAD activation drives apoptosis through a positive feedback loop, but also identify a unique suicide system that can be used for controlling gene-modified organisms. |
| Author | Ogawa, Hiromi Kanemaki, Masato T. Earnshaw, William C. Ageichik, Alexander V. Peterson, Kevin L. Kaufmann, Scott H. Samejima, Kumiko |
| Author_xml | – sequence: 1 givenname: Kumiko surname: Samejima fullname: Samejima, Kumiko email: kumiko.samejima@ed.ac.uk organization: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, Scotland, United Kingdom – sequence: 2 givenname: Hiromi surname: Ogawa fullname: Ogawa, Hiromi organization: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, Scotland, United Kingdom – sequence: 3 givenname: Alexander V. surname: Ageichik fullname: Ageichik, Alexander V. organization: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, Scotland, United Kingdom – sequence: 4 givenname: Kevin L. surname: Peterson fullname: Peterson, Kevin L. organization: Mayo Clinic, Rochester, Minnesota 55905 – sequence: 5 givenname: Scott H. surname: Kaufmann fullname: Kaufmann, Scott H. organization: Mayo Clinic, Rochester, Minnesota 55905 – sequence: 6 givenname: Masato T. surname: Kanemaki fullname: Kanemaki, Masato T. organization: Centre for Frontier Research, National Institute of Genetics, ROIS, and Department of Genetics, SOKENDAI, Yata 1111, Mishima, Shizuoka 411-8540, Japan, and – sequence: 7 givenname: William C. surname: Earnshaw fullname: Earnshaw, William C. email: bill.earnshaw@ed.ac.uk organization: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Max Born Crescent, Edinburgh EH9 3BF, Scotland, United Kingdom |
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| Title | Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death |
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