GSTpi reduces DNA damage and cell death by regulating the ubiquitination and nuclear translocation of NBS1

AbstractDuring Drosophila embryonic development, cell death eliminates 30% of the primordial germ cells (PGCs). Inhibiting apoptosis does not prevent PGC death, suggesting a divergence from the conventional apoptotic program. Here, we demonstrate that PGCs normally activate an intrinsic alternative cell death (ACD) pathway mediated by DNase II release from lysosomes, leading to nuclear translocation and subsequent DNA double-strand breaks (DSBs). DSBs activate the DNA damage-sensing enzyme, Poly(ADP-ribose) (PAR) polymerase-1 (PARP-1) and the ATR/Chk1 branch of the DNA damage response. PARP-1 and DNase II engage in a positive feedback amplification loop mediated by the release of PAR polymers from the nucleus and the nuclear accumulation of DNase II in an AIF- and CypA-dependent manner, ultimately resulting in PGC death. Given the anatomical and molecular similarities with an ACD pathway called parthanatos, these findings reveal a parthanatos-like cell death pathway active during Drosophila development.

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Campylobacter jejuni Cas9 Modulates the Transcriptome in Caco-2 Intestinal Epithelial Cells

Genes ◽

10.3390/genes11101193 ◽

2020 ◽

Vol 11 (10) ◽

pp. 1193

Author(s):

Chinmoy Saha ◽

Deborah Horst-Kreft ◽

Inez Kross ◽

Peter J. van der Spek ◽

Rogier Louwen ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Epithelial Cells ◽

Campylobacter Jejuni ◽

Nuclear Translocation ◽

Intestinal Epithelial Cells ◽

Nuclear Factor Κb ◽

Intestinal Epithelial ◽

Human Intestinal Epithelial Cells ◽

Cellular Pathways

The zoonotic human pathogen Campylobacter jejuni is known for its ability to induce DNA-damage and cell death pathology in humans. The molecular mechanism behind this phenomenon involves nuclear translocation by Cas9, a nuclease in C. jejuni (CjeCas9) that is the molecular marker of the Type II CRISPR-Cas system. However, it is unknown via which cellular pathways CjeCas9 drives human intestinal epithelial cells into cell death. Here, we show that CjeCas9 released by C. jejuni during the infection of Caco-2 human intestinal epithelial cells directly modulates Caco-2 transcriptomes during the first four hours of infection. Specifically, our results reveal that CjeCas9 activates DNA damage (p53, ATM (Ataxia Telangiectasia Mutated Protein)), pro-inflammatory (NF-κB (Nuclear factor-κB)) signaling and cell death pathways, driving Caco-2 cells infected by wild-type C. jejuni, but not when infected by a cas9 deletion mutant, towards programmed cell death. This work corroborates our previous finding that CjeCas9 is cytotoxic and highlights on a RNA level the basal cellular pathways that are modulated.

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Sevoflurane Exposure Induces Neuronal Cell Parthanatos Initiated by DNA Damage in the Developing Brain via an Increase of Intracellular Reactive Oxygen Species

Frontiers in Cellular Neuroscience ◽

10.3389/fncel.2020.583782 ◽

2020 ◽

Vol 14 ◽

Author(s):

Meihua Piao ◽

Yingying Wang ◽

Nan Liu ◽

Xuedong Wang ◽

Rui Chen ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Nuclear Translocation ◽

Neuronal Cell Death ◽

Neuronal Cell ◽

Volatile Anesthetics ◽

Intracellular Reactive Oxygen Species ◽

Developing Brain ◽

Oxygen Species ◽

Parp 1

The safety of volatile anesthetics in infants and young children has been drawing increasing concern due to its potential neurotoxicity in the developing brain. Neuronal death is considered a major factor associated with developmental neurotoxicity after exposure to volatile anesthetics sevoflurane, but its mechanism remains elusive. Parthanatos, a new type of programmed cell death, resulting from poly (ADP-ribose) polymerase 1 (PARP-1) hyperactivation in response to DNA damage, was found to account for the pathogenesis of multiple neurological disorders. However, the role of Parthanatos in sevoflurane-induced neonatal neuronal cell death has not been investigated. To test it, neuronal cells treated with 2, 4, and 8% sevoflurane for 6, 12, and 24 h and postnatal day 7 rats exposed to 2.5% sevoflurane for 6 h were used in the present study. Our results found sevoflurane exposure induced neuronal cell death, which was accompanied by PARP-1 hyperactivation, cytoplasmic polymerized ADP-ribose (PAR) accumulation, mitochondrial depolarization, and apoptosis-inducing factor (AIF) nuclear translocation in the neuronal cells and hippocampi of rats. Pharmacological or genetic inhibition of PAPR-1 significantly alleviated sevoflurane-induced neuronal cell death and accumulation of PAR polymer and AIF nuclear translocation, which were consistent with the features of Parthanatos. We observed in vitro and in vivo that sevoflurane exposure resulted in DNA damage, given that 8-hydroxydeoxyguanosine (8-OHdG) and phosphorylation of histone variant H2AX (γH2AX) were improved. Moreover, we detected that sevoflurane exposure was associated with an overproduction of intracellular reactive oxygen species (ROS). Inhibition of ROS with antioxidant NAC markedly alleviated DNA damage caused by sevoflurane, indicating that ROS participated in the regulation of sevoflurane-induced DNA damage. Additionally, sevoflurane exposure resulted in upregulation of Parthanatos-related proteins and neuronal cell death, which were significantly attenuated by pretreatment with NAC. Therefore, these results suggest that sevoflurane exposure induces neuronal cell Parthanatos initiated by DNA damage in the developing brain via the increase of intracellular ROS.

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NAMPT-derived NAD+ fuels PARP1 to promote skin inflammation through parthanatos cell death

PLoS Biology ◽

10.1371/journal.pbio.3001455 ◽

2021 ◽

Vol 19 (11) ◽

pp. e3001455

Author(s):

Francisco J. Martínez-Morcillo ◽

Joaquín Cantón-Sandoval ◽

Francisco J. Martínez-Navarro ◽

Isabel Cabas ◽

Idoya Martínez-Vicente ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Nuclear Translocation ◽

Skin Inflammation ◽

Ros Scavenging ◽

Nadph Oxidases ◽

Nicotinamide Phosphoribosyltransferase ◽

Pharmacological Inhibition ◽

Chronic Skin ◽

Apoptosis Inducing Factor

Several studies have revealed a correlation between chronic inflammation and nicotinamide adenine dinucleotide (NAD+) metabolism, but the precise mechanism involved is unknown. Here, we report that the genetic and pharmacological inhibition of nicotinamide phosphoribosyltransferase (Nampt), the rate-limiting enzyme in the salvage pathway of NAD+ biosynthesis, reduced oxidative stress, inflammation, and keratinocyte DNA damage, hyperproliferation, and cell death in zebrafish models of chronic skin inflammation, while all these effects were reversed by NAD+ supplementation. Similarly, genetic and pharmacological inhibition of poly(ADP-ribose) (PAR) polymerase 1 (Parp1), overexpression of PAR glycohydrolase, inhibition of apoptosis-inducing factor 1, inhibition of NADPH oxidases, and reactive oxygen species (ROS) scavenging all phenocopied the effects of Nampt inhibition. Pharmacological inhibition of NADPH oxidases/NAMPT/PARP/AIFM1 axis decreased the expression of pathology-associated genes in human organotypic 3D skin models of psoriasis. Consistently, an aberrant induction of NAMPT and PARP activity, together with AIFM1 nuclear translocation, was observed in lesional skin from psoriasis patients. In conclusion, hyperactivation of PARP1 in response to ROS-induced DNA damage, fueled by NAMPT-derived NAD+, mediates skin inflammation through parthanatos cell death.

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An absence of lamin B1 in migrating neurons causes nuclear membrane ruptures and cell death

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1917225116 ◽

2019 ◽

Vol 116 (51) ◽

pp. 25870-25879 ◽

Cited By ~ 11

Author(s):

Natalie Y. Chen ◽

Ye Yang ◽

Thomas A. Weston ◽

Jason N. Belling ◽

Patrick Heizer ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Nuclear Membrane ◽

Neuronal Migration ◽

Cortical Neurons ◽

Nuclear Translocation ◽

Neuronal Survival ◽

Ventricular Zone ◽

Cortical Plate ◽

Lamin B1

Deficiencies in either lamin B1 or lamin B2 cause both defective migration of cortical neurons in the developing brain and reduced neuronal survival. The neuronal migration abnormality is explained by a weakened nuclear lamina that interferes with nucleokinesis, a nuclear translocation process required for neuronal migration. In contrast, the explanation for impaired neuronal survival is poorly understood. We hypothesized that the forces imparted on the nucleus during neuronal migration result in nuclear membrane (NM) ruptures, causing interspersion of nuclear and cytoplasmic contents—and ultimately cell death. To test this hypothesis, we bredLmnb1-deficient mice that express a nuclear-localized fluorescentCrereporter. Migrating neurons within the cortical plate of E18.5Lmnb1-deficient embryos exhibited NM ruptures, evident by the escape of the nuclear-localized reporter into the cytoplasm and NM discontinuities by electron microscopy. The NM ruptures were accompanied by DNA damage and cell death. The NM ruptures were not observed in nonmigrating cells within the ventricular zone. NM ruptures, DNA damage, and cell death were also observed in culturedLmnb1−/−andLmnb2−/−neurons as they migrated away from neurospheres. To test whether mechanical forces on the cell nucleus are relevant to NM ruptures in migrating neurons, we examined culturedLmnb1−/−neurons when exposed to external constrictive forces (migration into a field of tightly spaced silicon pillars). As the cells entered the field of pillars, there were frequent NM ruptures, accompanied by DNA damage and cell death.

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GSTpi Reduces DNA Damage And Cell Death By Regulating The Ubiquitination And Nuclear Translocation of NBS1

10.21203/rs.3.rs-814053/v1 ◽

2021 ◽

Author(s):

Jinyi Zhou ◽

Lili Gu ◽

Yingying Shi ◽

Ting Huang ◽

Xirui Fan ◽

...

Keyword(s):

Dna Damage ◽

Cell Death ◽

Nuclear Translocation ◽

Nijmegen Breakage Syndrome ◽

Breast Adenocarcinoma ◽

Protection Mechanism ◽

Damage Level ◽

P53 Signaling ◽

M Phase ◽

Sensor Proteins

Abstract Glutathione S-transferase pi (GSTpi) is an important phase Ⅱ detoxifying enzyme that participates in various physiological processes, such as antioxidant, detoxification, and signal transduction. The high expression level of GSTpi has been reported to be related to drug-resistant and anti-inflammatory and it functioned via its non-catalytic ligandin. However, the previous protection mechanism of GSTpi in DNA damage has not been addressed so far. Nijmegen breakage syndrome 1 (NBS1) is one of the most important sensor proteins to detect damaged DNA. Here, we investigated the interaction between GSTpi and NBS1 in HEK-293T cells and human breast adenocarcinoma cells during DNA damage. Our results showed that overexpression of GSTpi in cells by transfecting DNA vector decreased the DNA damage level after Methyl methanesulfonate (MMS) or Adriamycin (ADR) treatment. We found that cytosolic GSTpi could increase NBS1 ubiquitin-mediated degradation in unstimulated cells, which suggested that GSTpi could maintain the basal level of NBS1 during normal conditions. In response to DNA damage, GSTpi can be phosphorylated in Ser184 and inhibit the ubiquitination degradation of NBS1 mediated by Skp2 to recover NBS1 protein level. Phosphorylated GSTpi can further enhance NBS1 nuclear translocation to activate the ATM-Chk2-p53 signaling pathway. Finally, GSTpi blocked the cell cycle in the G2/M phase to gain more time for cells to repair. Thus, our finding revealed the novel mechanism of GSTpi via its Ser184 phosphorylation to protect cells from cell death during DNA damage and it enriches the function of GSTpi in drug resistance.

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