Apoptotic DNA Degradation into Oligonucleosomal Fragments, but Not Apoptotic Nuclear Morphology, Relies on a Cytosolic Pool of DFF40/CAD Endonuclease

Iglesias-Guimarais, Victoria; Gil-Guiñón, Estel; Gabernet, Gisela; Garcia-Belinchón, Mercè; Sánchez-Osuna, María; Casanelles, Elisenda; Comella, Joan X.; Yuste, Vı́ctor J.

doi:10.1074/jbc.m111.290718

Cited by 29 publications

(33 citation statements)

References 52 publications

(66 reference statements)

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“…More recently, we have identified SK-N-AS cells as an interesting cellular model to elucidate how stage II apoptotic nuclear morphology takes place. Indeed, these cells hold a defect in the hydrolysis of their chromatin into oligonucleosomal-size fragments, albeit the presence of stage II apoptotic nuclei upon cytotoxic insult (22). We show here that DFF40/CAD knockdown in SK-N-AS cells completely avoids caspase-dependent nuclear collapse and disassembly without affecting the initial ring-like marginalization of the chromatin inside the nucleus (stage I).…”

Section: Discussionmentioning

confidence: 75%

“…The degree of DFF40/CAD activation is directly related to the proteolytic action of caspases, and its enzymatic activity is further regulated by other factors, including histone H1 (11,47), HMG-1 (11), CIIA (48), or nucleophosmin/B23 (49). On the other hand, we have recently described that SK-N-AS cells show higher DFF40/CAD protein amounts than IMR-5 cells but less than SH-SY5Y cells (22). Therefore, it should be plausible that stage II nuclear morphology (mediated by the induction of ssDNA nicks/breaks) or DNA laddering require different amounts of DFF40/CAD pro- tein for it to take place.…”

Section: Discussionmentioning

confidence: 99%

“…Even in the Absence of Oligonucleosomal DNA FragmentationWe have recently identified SK-N-AS cells, a human neuroblastoma-derived cell line, as a cellular model that undergoes incomplete apoptotic cell death after cytotoxic stimuli, characterized by the presence of apoptotic nuclei in the absence of DNA laddering (22). First, we established that the aspect of these nuclei depends on the activation of caspases (supplemental Fig.…”

Section: Dff40/cad Is Required For Stage II Nuclear Morphologymentioning

confidence: 94%

“…Low Molecular Weight DNA Degradation Analysis-The DNA fragmentation assays were carried out as described previously (22). DNA was stained in ethidium bromide and then visualized using a Syngene Gene Genius UV transilluminator equipped with a photographic camera.…”

Section: Methodsmentioning

confidence: 99%

“…They undergo an incomplete caspase-dependent apoptosis with highly compacted chromatin in the absence of DNA laddering (22). Finding such apoptotic behavior should provide new insights on how the final apoptotic chromatin compaction takes place and whether DFF40/CAD plays a role in this process.…”

mentioning

confidence: 99%

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Chromatin Collapse during Caspase-dependent Apoptotic Cell Death Requires DNA Fragmentation Factor, 40-kDa Subunit-/Caspase-activated Deoxyribonuclease-mediated 3′-OH Single-strand DNA Breaks

Iglesias-Guimarais

Gil-Guiñón

Sánchez-Osuna

et al. 2013

Journal of Biological Chemistry

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Background: Apoptotic nuclear morphology can occur independently of DFF40/CAD-mediated DNA fragmentation. Results: DFF40/CAD induces 3Ј-OH single-strand DNA nicks/breaks and nuclear collapse during caspase-dependent apoptosis. Conclusion: Caspase-dependent apoptotic nuclear collapse is prompted by DFF40/CAD-mediated single-strand DNA damage. Significance: The knowledge of how apoptotic nuclear collapse occurs should be relevant to understand the final steps of cell demise and its influence on the cellular environment.

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Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Dff40/cad Is Required For Stage II Nuclear Morphologymentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Chromatin Collapse during Caspase-dependent Apoptotic Cell Death Requires DNA Fragmentation Factor, 40-kDa Subunit-/Caspase-activated Deoxyribonuclease-mediated 3′-OH Single-strand DNA Breaks

Iglesias-Guimarais

Gil-Guiñón

Sánchez-Osuna

et al. 2013

Journal of Biological Chemistry

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Toxicological Analysis UsingDNAFragmentation

Basnakian

Braman

Yin

et al. 2017

Encyclopedia of Analytical Chemistry

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Toxicological analysis relies on the measurement of cell death. DNA fragmentation is the preferential method in cell death assessment because it measures irreversible stage of cell death, and it is universal to all mechanisms of cell death. DNA fragmentation is provided by a group of nine cytotoxic DNases/endonucleases, cumulative activity of which is sufficient to eventually destroy all cellular DNAs to potentially reusable mononucleotides. Of all the methods for measuring DNA fragmentation, the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay seems to be the best in satisfying the toxicology assessment because it is simple, quantitative, based on individual cell measurements, and is not destructive to the cells. The latter allows it to combine with quantitative immunocytochemistry and other histology and genetic techniques to perform mechanistic studies.

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N‐hexane inhalation during pregnancy alters DNA promoter methylation in the ovarian granulosa cells of rat offspring

Liu

Sun³

et al. 2013

J of Applied Toxicology

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The N-hexane-induced impact on the reproductive system of the offspring of animals exposed to n-hexane has caused great concern. Pregnant Wistar rats inhaled 500, 2 500 or 12 500 ppm n-hexane during gestational days 1-20. Clinical characteristics and developmental indices were observed. Ovarian granulosa cells were extracted from F1 rats, the number of follicles was determined in ovarian slices and promoter methylation was assessed using MeDIP-Chip. Several methods were used to analyze the scanned genes, including the Gene Ontology Consortium tools, the DAVID Functional Annotation Clustering Tool, hierarchical clustering and KEGG pathway analysis. The results indicated that the live pups/litter ratio was significantly lowest in the 12 500 ppm group. A significant decrease in secondary follicles and an increase in atresic follicles were observed in the 12 500 ppm group. The number of shared demethylated genes was higher than that of the methylated genes, and the differentially methylated genes were enriched in cell death and apoptosis, cell growth and hormone regulation. The methylation profiles of the offspring from the 500 ppm and control groups were different from those of the 2500 and 12 500 ppm groups. Furthermore, the methylation status of genes in the PI3K-Akt and NF-kappa B signaling pathways was changed after n-hexane exposure. The Cyp11a1, Cyp17a1, Hsd3b1, Cyp1a1 and Srd5a1 promoters were hypermethylated in the n-hexane-exposed groups. These results indicate that the developmental toxicity of n-hexane in F1 ovaries is accompanied by the altered methylation of promoters of genes associated with apoptotic processes and steroid hormone biosynthesis.

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Apoptotic DNA Degradation into Oligonucleosomal Fragments, but Not Apoptotic Nuclear Morphology, Relies on a Cytosolic Pool of DFF40/CAD Endonuclease

Cited by 29 publications

References 52 publications

Chromatin Collapse during Caspase-dependent Apoptotic Cell Death Requires DNA Fragmentation Factor, 40-kDa Subunit-/Caspase-activated Deoxyribonuclease-mediated 3′-OH Single-strand DNA Breaks

Chromatin Collapse during Caspase-dependent Apoptotic Cell Death Requires DNA Fragmentation Factor, 40-kDa Subunit-/Caspase-activated Deoxyribonuclease-mediated 3′-OH Single-strand DNA Breaks

Toxicological Analysis UsingDNAFragmentation

N‐hexane inhalation during pregnancy alters DNA promoter methylation in the ovarian granulosa cells of rat offspring

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