RESUMO
Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in the pentose phosphate pathway (PPP) and plays an essential role in the oxidative stress response by producing NADPH, the main intracellular reductant. G6PD deficiency is the most common human enzyme defect, affecting more than 400 million people worldwide. Here, we show that G6PD is negatively regulated by acetylation on lysine 403 (K403), an evolutionarily conserved residue. The K403 acetylated G6PD is incapable of forming active dimers and displays a complete loss of activity. Knockdown of G6PD sensitizes cells to oxidative stress, and re-expression of wild-type G6PD, but not the K403 acetylation mimetic mutant, rescues cells from oxidative injury. Moreover, we show that cells sense extracellular oxidative stimuli to decrease G6PD acetylation in a SIRT2-dependent manner. The SIRT2-mediated deacetylation and activation of G6PD stimulates PPP to supply cytosolic NADPH to counteract oxidative damage and protect mouse erythrocytes. We also identified KAT9/ELP3 as a potential acetyltransferase of G6PD. Our study uncovers a previously unknown mechanism by which acetylation negatively regulates G6PD activity to maintain cellular NADPH homeostasis during oxidative stress.
Assuntos
Sobrevivência Celular/fisiologia , Glucosefosfato Desidrogenase/metabolismo , Histona Acetiltransferases/metabolismo , Homeostase/fisiologia , NADP/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Estresse Oxidativo/fisiologia , Sirtuína 2/metabolismo , Acetilação , Animais , Técnicas de Silenciamento de Genes , Glucosefosfato Desidrogenase/genética , Proteínas de Fluorescência Verde , Células HEK293 , Humanos , Camundongos , RNA Interferente Pequeno/genéticaRESUMO
The TET2 DNA dioxygenase regulates gene expression by catalyzing demethylation of 5-methylcytosine, thus epigenetically modulating the genome. TET2 does not contain a sequence-specific DNA-binding domain, and how it is recruited to specific genomic sites is not fully understood. Here we carried out a mammalian two-hybrid screen and identified multiple transcriptional regulators potentially interacting with TET2. The SMAD nuclear interacting protein 1 (SNIP1) physically interacts with TET2 and bridges TET2 to bind several transcription factors, including c-MYC. SNIP1 recruits TET2 to the promoters of c-MYC target genes, including those involved in DNA damage response and cell viability. TET2 protects cells from DNA damage-induced apoptosis dependending on SNIP1. Our observations uncover a mechanism for targeting TET2 to specific promoters through a ternary interaction with a co-activator and many sequence-specific DNA-binding factors. This study also reveals a TET2-SNIP1-c-MYC pathway in mediating DNA damage response, thereby connecting epigenetic control to maintenance of genome stability.
Assuntos
Dano ao DNA/genética , Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Biocatálise/efeitos dos fármacos , Linhagem Celular Tumoral , Cisplatino/farmacologia , Quebras de DNA de Cadeia Dupla , Proteínas de Ligação a DNA/química , Dioxigenases , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Ligação Proteica/efeitos dos fármacos , Proteínas Proto-Oncogênicas/química , Proteínas de Ligação a RNA , Transcrição Gênica/efeitos dos fármacosRESUMO
Based on the long-term fertilization experiments, effects of various fertilization practices on the soil organic carbon (SOC) and total nitrogen (TN) in the surface (0-20 cm) and subsurface (20-40 cm) black soil in northeast China were studied. Results showed that, compared with the CK, long-term application of organic manure, especially the combination of mineral fertilizers and organic manure significantly increased the organic SOC and TN in the surface soil. Application of mineral fertilizers plus organic manure with conventional (NPM) and high application (N2P2M2) rate increased SOC significantly by 24. 6% and 25.1% , and TN by 29.5% and 32.8%, respectively. However, there was no significant difference among the treatments for SOC and TN at the subsurface. Compared with the CK (CKh), mineral fertilizer plus organic manure (NPM and N2P2M2) did not only increase the soil microbial biomass carbon (SMBC) and nitrogen (SMBN) , dissolved organic carbon (DOC) and nitrogen (DN), but also significantly increased the ratio of SMBC and DOC to SOC, SMBN and TN to TN. Application of the NPM and N2P2M2 increased the value of SMBC/SOC by 0.36 to 0.59 and SMBN/TN by 1.21 to 1.95 percentage points, respectively. The value of DOC/SOC and DN/TN ranged from 0.53% to 0.72% and 1.41% to 1.78%, respectively. This result indicated that SMBC, SMBN, DOC, DN and SMBC/ SOC, SMBN/TN, DOC/SOC, DN/TN were more sensitive than SOC and TN to long-term fertilization in the soil profile, and were better indicators for the impact of long-term fertilization soil fertility. The concluded that the application of manure especially manure plus mineral fertilizers can increase soil nutrients activity in the surface and subsurface black soil, acting as a helpful practice to improve soil fertility and the ability of nutrient supply, while it may cause potential environment pollution on carbon and nitrogen loss in the agroecosystem.
Assuntos
Carbono/análise , Fertilizantes , Nitrogênio/análise , Compostos Orgânicos/análise , Solo/química , China , Produtos Agrícolas/crescimento & desenvolvimento , Monitoramento Ambiental , Fatores de TempoRESUMO
In order to explore the effects of long-term fertilization on the microbiological characters of red soil, soil samples were collected from a 19-year long-term experimental field in Qiyang of Hunan, with their microbial biomass carbon (MBC) and nitrogen (MBN) and microbial utilization ratio of carbon sources analyzed. The results showed that after 19-year fertilization, the soil MBC and MBN under the application of organic manure and of organic manure plus inorganic fertilizers were 231 and 81 mg x kg(-1) soil, and 148 and 73 mg x kg(-1) soil, respectively, being significantly higher than those under non-fertilization, inorganic fertilization, and inorganic fertilization plus straw incorporation. The ratio of soil MBN to total N under the application of organic manure and of organic manure plus inorganic fertilizers was averagely 6.0%, significantly higher than that under non-fertilization and inorganic fertilization. Biolog-ECO analysis showed that the average well color development (AWCD) value was in the order of applying organic manure plus inorganic fertilizers = applying organic manure > non-fertilization > inorganic fertilization = inorganic fertilization plus straw incorporation. Under the application of organic manure or of organic manure plus inorganic fertilizers, the microbial utilization rate of carbon sources, including carbohydrates, carboxylic acids, amino acids, polymers, phenols, and amines increased; while under inorganic fertilization plus straw incorporation, the utilization rate of polymers was the highest, and that of carbohydrates was the lowest. Our results suggested that long-term application of organic manure could increase the red soil MBC, MBN, and microbial utilization rate of carbon sources, improve soil fertility, and maintain a better crop productivity.
Assuntos
Carbono/análise , Fertilizantes , Nitrogênio/análise , Microbiologia do Solo , Solo/análise , Agricultura/métodos , Fatores de Tempo , Triticum/crescimento & desenvolvimentoRESUMO
Exited states in 134Pr were populated in the fusion-evaporation reaction 119Sn(19F,4n)134Pr. Recoil distance Doppler-shift and Doppler-shift attenuation measurements using the Euroball spectrometer, in conjunction with the inner Bismuth Germanate ball and the Cologne plunger, were performed at beam energies of 87 MeV and 83 MeV, respectively. Reduced transition probabilities in 134Pr are compared to the predictions of the two quasiparticle + triaxial rotor and interacting boson fermion-fermion models. The experimental results do not support the presence of static chirality in 134Pr underlying the importance of shape fluctuations. Only within a dynamical context the presence of intrinsic chirality in 134Pr can be supported.
RESUMO
We present a simple method for discerning the evolution from vibrational to rotational structure in nuclei as a function of spin. The prescription is applied to the yrast cascades in the A approximately 110 region and a clear transition from vibrational to rotational motion is found.