Mapping the initial DNA breaks in apoptotic Jurkat cells using ligation-mediated PCR

Liu, Q Y; Ribecco-Lutkiewicz, Maria; Carson, Christine; Testolin, L.; Bergeron, Dominique; Kohwi‐Shigematsu, Terumi; Walker, P. Roy; Sikorska, Marianna

doi:10.1038/sj.cdd.4401146

Cited by 22 publications

(36 citation statements)

References 54 publications

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“…Total cellular and nuclear proteins were extracted as described by Liu et al 17 The cytoskeletal structures were removed from the nuclear pellet by washing with cytoskeletal stripping buffer (BNB buffer containing a 2 : 1 ratio (v/v) of NP-40 and sodium deoxycholate in a final concentration of 1% : 0.5%) for 15 min and centrifugation at 3000 r.p.m. for 5 min at 41C.…”

Section: Protein Extractionmentioning

confidence: 99%

“…This results in the generation of single-strand breaks (ssb) and, subsequently, double-strand (ds) DNA fragmentation. [15][16][17] Recent reports imply that DNaseY is activated by an increase of intracellular Ca 2 þ , which results in early DNA breaks and activation of PARP-1. Poly-ADP-ribosylation of DNaseY by PARP-1 inhibits its endonuclease activity.…”

Section: Introductionmentioning

confidence: 99%

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Regulation of DNaseY activity by actinin-α4 during apoptosis

et al. 2004

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DNaseY, a Ca 2 þ -and Mg 2 þ -dependent endonuclease, has been implicated in apoptotic DNA degradation; however, the molecular mechanisms controlling its involvement in this process have not been fully elucidated. We have obtained evidence from yeast two-hybrid screening and coimmunoprecipitation experiments that DNaseY interacted physically with actinin-a4 and this interaction significantly enhanced its endonuclease activity. Accordingly, simultaneous overexpression of both proteins in PC12 cells dramatically increased the rate of apoptosis in response to teniposide' VM26. However, overexpression of DNaseY alone neither triggered apoptosis nor facilitated cell death in response to VM26 or serum deprivation. Instead, the overexpression of DNaseY increased the production of single-strand DNA breaks and evoked a profound upregulation of DNA repair pathways. Taken together, our results point to a novel regulatory mechanism of DNaseY activity and offer an explanation for why cells must first cleave key DNA repair and replication proteins before the successful execution of apoptosis.

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Section: Protein Extractionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Regulation of DNaseY activity by actinin-α4 during apoptosis

et al. 2004

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“…Examination of the myogenin promoter showed no enrichment for CAD at these time points. To localize the formation of strand breaks in the p21 promoter, we used a modified ligation-mediated PCR (LM-PCR) protocol; LM-PCR has been used to map DNA strand breaks that occur during apoptosis as well as site-specific strand breaks that result from glucocoticoid stimulation of Michigan Cancer Foundation (MCF)-7 cells (14,27). Briefly, the DNA linker is ligated to a DNA strand break.…”

mentioning

confidence: 99%

“…Our observations implicate caspase/CAD induction of p21 gene expression, yet the pattern of strand-break formation and repair (ISNT-labeled foci and phospho-H2AX) during myoblast differentiation is consistent with a genome-wide modification rather than a targeting of a single loci. Previous studies have linked CAD targeting to regions of DNA that have equitable distribution, such as accessible intranucleosomal linker regions and/ or matrix attachment regions (MARs) (10,27). Depending on the context of these targeted regions, one would predict that the CADdirected cleavage events may be repressive for some genes/genomic loci but inductive for others.…”

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confidence: 99%

Caspase 3/caspase-activated DNase promote cell differentiation by inducing DNA strand breaks

Larsen

Rampalli

Burns

et al. 2010

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

205

204

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Caspase 3 is required for the differentiation of a wide variety of cell types, yet it remains unclear how this apoptotic protein could promote such a cell-fate decision. Caspase signals often result in the activation of the specific nuclease caspase-activated DNase (CAD), suggesting that cell differentiation may be dependent on a CADmediated modification in chromatin structure. In this study, we have investigated if caspase 3/CAD plays a role in initiating the DNA strand breaks that are known to occur during the terminal differentiation of skeletal muscle cells. Here, we show that inhibition of caspase 3 or reduction of CAD expression leads to a dramatic loss of strand-break formation and a block in the myogenic program. Caspase-dependent induction of differentiation results in CAD targeting of the p21 promoter, and loss of caspase 3 or CAD leads to a significant down-regulation in p21 expression. These results show that caspase 3/CAD promotes cell differentiation by directly modifying the DNA/nuclear microenvironment, which enhances the expression of critical regulatory genes.development | gene expression | non-apoptotic caspase activity | epigenetics

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“…After liquefaction of semen, at room temperature, standard semen parameters were obtained according to WHO [2010] guidelines. Sperm viability was assessed 30 min after ejaculation [30][31][32][33][34][35][36].…”

Section: Semen Analysismentioning

confidence: 99%

Effect of Seminal Transforming Growth Factorß1 (TGFß1) and Glutathione on Apoptosis in Spermatozoa from Tunisian Infertile Men

Ben Ali

2017

Andrology

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We aimed to compare the effect of TGFB1 and glutathione on sperm necrosis in three groups of patients with increased necrospermia. The study included 120 men aged 37.6 ± 4.6 years consulting for sterility of the couple. Patients were subdivided into three groups according to the percentage of necrotic spermatozoa: necrospermia<30%; N=40), moderate necrozoospermia (50-80%, n=45) and severe necrozoospermia (>80%) For each patient, the sperm parameters were evaluated according to WHO standards, TGFB1 and glutathione Oxidized and reduced was carried out by the ELISA technique and by spectrophotometry respectively. We found a significant increase in the levels of TGFβ1 and DFI in moderate and severe necrospermia groups and positive correlations between these two factors and sperm parameters (numeration, mobility and morphology). In addition, a significant decrease in seminal GHSt was observed in the necrozoospermic groups compared to the control group. A strong correlation was observed between the degree of necrozoospermia and the fragmentation of sperm DNA (r=0.878, p=0.001). In addition, a significant positive correlation between TGFβ1 and DFI in the two groups of patients with moderate necrospermia (r=0.43, p<0.05) and severe necrospermia (r=0.52, p<0.05). This confirms that seminal TGFβ1 is a factor in the fragmentation of spermatozoa DNA. These results suggest that decreased glutathione and increased seminal TGFβ1 are important risk factors that can lead to a succession of morphological and functional alterations of spermatozoa resulting in necrozoospermia. Their perturbation in seminal plasma can lead to mediocre results of medically assisted procreation.

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Mapping the initial DNA breaks in apoptotic Jurkat cells using ligation-mediated PCR

Cited by 22 publications

References 54 publications

Regulation of DNaseY activity by actinin-α4 during apoptosis

Regulation of DNaseY activity by actinin-α4 during apoptosis

Caspase 3/caspase-activated DNase promote cell differentiation by inducing DNA strand breaks

Effect of Seminal Transforming Growth Factorß1 (TGFß1) and Glutathione on Apoptosis in Spermatozoa from Tunisian Infertile Men

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