Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death

Samejima, Kumiko; Ogawa, Hiromi; Ageichik, Alexander V.; Peterson, Kevin; Kaufmann, Scott H.; Kanemaki, Masato T.; Earnshaw, William C.

doi:10.1074/jbc.m114.583542

Cited by 29 publications

(23 citation statements)

References 36 publications

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“…; Samejima et al . ). Taking advantage of CRISPR–Cas9‐based gene editing technology, which allows efficient knock‐in at a targeted locus (Cong et al .…”

Section: Resultsmentioning

confidence: 97%

Perichromosomal protein Ki67 supports mitotic chromosome architecture

et al. 2016

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Although the condensin complexes and topoisomerase IIa (TopoIIa) are the central players in mitotic chromosome formation, they are insufficient for its completion, and additional factors involved in the process have been extensively sought. In this study, we examined the possibility that Ki67, a perichromosomal protein widely used as a cell proliferation marker, is one such factor. Using a combination of auxin-inducible degron and CRISPR-Cas9-based gene editing technologies, we generated a human HCT116 cell line in which Ki67 is rapidly depleted in a few hours. The removal of Ki67 before mitotic entry did not impact the early mitotic chromosome assembly observed in prophase but subsequently resulted in the formation of misshapen mitotic chromosomes. When Ki67 was removed after mitotic entry, preassembled rod-shaped mitotic chromosomes became disorganized. In addition, we show that Ki67 and TopoIIa are reciprocally coimmunoprecipitated from mitotic cell extracts. These observations indicate that Ki67 aids the finalization of mitotic chromosome formation and helps maintain rod-shaped chromosome architecture, likely in collaboration with TopoIIa. Together, these findings represent a new model in which mitotic chromosome architecture is supported both internally and externally.

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“…; Samejima et al . ). Taking advantage of CRISPR–Cas9‐based gene editing technology, which allows efficient knock‐in at a targeted locus (Cong et al .…”

Section: Resultsmentioning

confidence: 97%

Perichromosomal protein Ki67 supports mitotic chromosome architecture

et al. 2016

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“…Because yeast lack a traditional apoptotic pathway, caspases, and the ICAD/CAD node, the galactose inducible TEVP strategy only proved useful to study chromatin structure in this context. More recently, Samejima et al described a cellular strategy that relied on an auxin-inducible degron system for inducing rapid and complete proteasomal degradation of huICAD in double-knockout DT40 chicken cells (39). It remains unclear how authentic their strategy is for selective CAD activation because it does not recapitulate both the spatial and endogenous site-specific mechanism of activation.…”

Section: Discussionmentioning

confidence: 99%

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

Morgan

Diaz

Zeitlin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Cellular demolition during apoptosis is completed by executioner caspases, that selectively cleave more than 1,500 proteins but whose individual roles are challenging to assess. Here, we used an optimized site-specific and inducible protease to examine the role of a classic apoptotic node, the caspase-activated DNase (CAD). CAD is activated when caspases cleave its endogenous inhibitor ICAD, resulting in the characteristic DNA laddering of apoptosis. We describe a posttranscriptional gene replacement (PTGR) approach where endogenous biallelic ICAD is knocked down and simultaneously replaced with an engineered allele that is susceptible to inducible cleavage by tobacco etch virus protease. Remarkably, selective activation of CAD alone does not induce cell death, although hallmarks of DNA damage are detected in human cancer cell lines. Our data strongly support that the highly cooperative action of CAD and inhibition of DNA repair systems are critical for the DNA laddering phenotype in apoptosis. Furthermore, the PTGR approach provides a general means for replacing wild-type protein function with a precisely engineered mutant at the transcriptional level that should be useful for cell engineering studies.DNA damage | apoptosis | ICAD | site-specific proteolysis |

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“…This system is called the auxininducible degron system (Nishimura et al, 2009;Holland et al, 2012;Kanke et al, 2012;Farr et al, 2014;Nishimura and Kanemaki, 2014;Samejima et al, 2014). All eukaryotes possess SCF ubiquitin ligases, and the architecture of Arabidopsis TIR1, including the F-box domain, is sufficiently conserved to allow assembly into an SCF TIR1 complex in yeast and animals.…”

Section: Use Of Auxin-inducible Degrons As a Tool In Animal Systemsmentioning

confidence: 99%

SCF^TIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development

2015

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Auxin regulates a vast array of growth and developmental processes throughout the life cycle of plants. Auxin responses are highly context dependent and can involve changes in cell division, cell expansion, and cell fate. The complexity of the auxin response is illustrated by the recent finding that the auxin-responsive gene set differs significantly between different cell types in the root. Auxin regulation of transcription involves a core pathway consisting of the TIR1/AFB F-box proteins, the Aux/IAA transcriptional repressors, and the ARF transcription factors. Auxin is perceived by a transient coreceptor complex consisting of a TIR1/AFB protein and an Aux/IAA protein. Auxin binding to the coreceptor results in degradation of the Aux/ IAAs and derepression of ARF-based transcription. Although the basic outlines of this pathway are now well established, it remains unclear how specificity of the pathway is conferred. However, recent results, focusing on the ways that these three families of proteins interact, are starting to provide important clues.

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Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death

Cited by 29 publications

References 36 publications

Perichromosomal protein Ki67 supports mitotic chromosome architecture

Perichromosomal protein Ki67 supports mitotic chromosome architecture

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

SCF^TIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development

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Auxin-induced Rapid Degradation of Inhibitor of Caspase-activated DNase (ICAD) Induces Apoptotic DNA Fragmentation, Caspase Activation, and Cell Death

Cited by 29 publications

References 36 publications

Perichromosomal protein Ki67 supports mitotic chromosome architecture

Perichromosomal protein Ki67 supports mitotic chromosome architecture

Engineered cellular gene-replacement platform for selective and inducible proteolytic profiling

SCFTIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development

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SCF^TIR1/AFB-Based Auxin Perception: Mechanism and Role in Plant Growth and Development