The programmed necrosis induced by TNF-α requires the activities of the receptor-interacting serine-threonine kinases RIP1 and RIP3 and their interaction with the mixed lineage kinase domain-like ...protein MLKL. We report the identification of RIP1- and RIP3-containing protein complexes that form specifically in response to necrosis induction. One component of these complexes is the mitochondrial protein phosphatase PGAM5, which presents as two splice variants, PGAM5L (long form) and PGAM5S (short form). Knockdown of either form attenuated necrosis induced by TNF-α as well as reactive oxygen species (ROS) and calcium ionophore, whereas knockdown of RIP3 and MLKL blocked only TNF-α-mediated necrosis. Upon necrosis induction, PGAM5S recruited the mitochondrial fission factor Drp1 and activated its GTPase activity by dephosphorylating the serine 637 site of Drp1. Drp1 activation caused mitochondrial fragmentation, an early and obligatory step for necrosis execution. These data defined PGAM5 as the convergent point for multiple necrosis pathways.
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► PGAM5 is a mitochondrial phosphatase that functions in programmed necrosis (necroptosis) ► Both splicing variants of PGAM5, PGAM5L and PGAM5S, are required for necroptosis ► PGAM5 dephosphorylates and activates mitochondrial fission protein Drp1 ► PGAM5 is required for necrosis induced by TNF-α, ROS, and calcium overload
PGAM5 is the downstream target of the necrosome consisting of receptor-interacting kinases RIP1, RIP3, and MLKL. PGAM5 dephosphorylates and activates the mitochondrial fission factor Drp1 to cause mitochondrial fragmentation, an early and obligatory step for necrosis execution.
Legacy organic contaminants, such as polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), and several currently used organic contaminants, such as novel brominated retardants ...(NBFRs), organophosphate flame retardants (OPFRs), and Dechlorane Plus (DPs), were measured in the segmented hair and hand wipes of e-waste dismantling workers and in the dust from their workplace in an e-waste recycling site in South China to assess the accumulation and sources of organic contaminants in the hair shaft. The median concentrations of DPs, PBDEs, NBFRs, PCBs and OPFRs in hair samples were 23.5, 154, 156, 349, and 371 ng/g, respectively. A linear increase in organic contaminant concentrations was found along the hair shaft, with significant differences among each segment, while the age-related patterns were similar among the 7 hair segments. A linear relationship was found between the accumulation rates and the log K OW, indicating that the accumulation rates of organic contaminants along the hair shaft decreased with increasing log K OW values. Enantiomer fraction (EF) values of PCB-132 in distal segments were close to those in dust and air, while EFs in proximal segments were close to those in serum. The contributions of PCBs from external sources to hair increased with the distance from the scalp of hair segments, from 67.8% in 0–3 cm segments to 95.9% in 18–21 cm segments.
Calcium is a universal secondary messenger that triggers many cellular responses. However, it is unclear how a calcium signal is coordinately decoded by different calcium sensors, which in turn ...regulate downstream targets to fulfill a specific physiological function. Here we show that SOS2-LIKE PROTEIN KINASE5 (PKS5) can negatively regulate the Salt-Overly-Sensitive signaling pathway in Arabidopsis. PKS5 can interact with and phosphorylate SOS2 at Ser
, promote the interaction between SOS2 and 14-3-3 proteins, and repress SOS2 activity. However, salt stress promotes an interaction between 14-3-3 proteins and PKS5, repressing its kinase activity and releasing inhibition of SOS2. We provide evidence that 14-3-3 proteins bind to Ca
, and that Ca
modulates 14-3-3-dependent regulation of SOS2 and PKS5 kinase activity. Our results suggest that a salt-induced calcium signal is decoded by 14-3-3 and SOS3/SCaBP8 proteins, which selectively activate/inactivate the downstream protein kinases SOS2 and PKS5 to regulate Na
homeostasis by coordinately mediating plasma membrane Na
/H
antiporter and H
-ATPase activity.
Codon usage biases are found in all eukaryotic and prokaryotic genomes, and preferred codons are more frequently used in highly expressed genes. The effects of codon usage on gene expression were ...previously thought to be mainly mediated by its impacts on translation. Here, we show that codon usage strongly correlates with both protein and mRNA levels genome-wide in the filamentous fungus Neurospora. Gene codon optimization also results in strong up-regulation of protein and RNA levels, suggesting that codon usage is an important determinant of gene expression. Surprisingly, we found that the impact of codon usage on gene expression results mainly from effects on transcription and is largely independent of mRNA translation and mRNA stability. Furthermore, we show that histone H3 lysine 9 trimethylation is one of the mechanisms responsible for the codon usage-mediated transcriptional silencing of some genes with nonoptimal codons. Together, these results uncovered an unexpected important role of codon usage in ORF sequences in determining transcription levels and suggest that codon biases are an adaptation of protein coding sequences to both transcription and translation machineries. Therefore, synonymous codons not only specify protein sequences and translation dynamics, but also help determine gene expression levels.
During autophagosome formation in mammalian cells, isolation membranes (IMs; autophagosome precursors) dynamically contact the ER. Here, we demonstrated that the ER-localized metazoan-specific ...autophagy protein EPG-3/VMP1 controls ER-IM contacts. Loss of VMP1 causes stable association of IMs with the ER, thus blocking autophagosome formation. Interaction of WIPI2 with the ULK1/FIP200 complex and PI(3)P contributes to the formation of ER-IM contacts, and these interactions are enhanced by VMP1 depletion. VMP1 controls contact formation by promoting SERCA (sarcoendoplasmic reticulum calcium ATPase) activity. VMP1 interacts with SERCA and prevents formation of the SERCA/PLN/SLN inhibitory complex. VMP1 also modulates ER contacts with lipid droplets, mitochondria, and endosomes. These ER contacts are greatly elevated by the SERCA inhibitor thapsigargin. Calmodulin acts as a sensor/effector to modulate the ER contacts mediated by VMP1/SERCA. Our study provides mechanistic insights into the establishment and disassociation of ER-IM contacts and reveals that VMP1 modulates SERCA activity to control ER contacts.
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•The ER-localized autophagy protein VMP1 modulates ER contact maintenance/disassembly•WIPI2 interacts with the ULK1/FIP200 complex and PI(3)P for ER-IM contact formation•VMP1 controls membrane contact disassembly by promoting SERCA activity•VMP1 prevents formation of the SERCA/PLN/SLN inhibitory complex
Zhao et al. demonstrate that the ER-localized metazoan-specific autophagy protein VMP1 controls ER contacts with IMs and other organelles. VMP1 controls contact maintenance by modulating SERCA activity. VMP1 interacts with SERCA and prevents formation of the SERCA/PLN/SLN inhibitory complex.
Autophagy mediates the cellular response to nutrient deprivation, protein aggregation, and pathogen invasion in human. Dysfunction of autophagy has been implicated in multiple human diseases ...including cancer. The identification of novel autophagy factors in mammalian cells will provide critical mechanistic insights into how this complicated cellular pathway responds to a broad range of challenges. Here, we report the cloning of an autophagy-specific protein that we called Barkor (Beclin 1-associated autophagy-related key regulator) through direct interaction with Beclin 1 in the human phosphatidylinositol 3-kinase class III complex. Barkor shares 18% sequence identity and 32% sequence similarity with yeast Atg14. Elimination of Barkor expression by RNA interference compromises starvation- and rapamycin-induced LC3 lipidation and autophagosome formation. Overexpression of Barkor leads to autophagy activation and increased number and enlarged volume of autophagosomes. Tellingly, Barkor is also required for suppression of the autophagy-mediated intracellular survival of Salmonella typhimurium in mammalian cells. Mechanistically, Barkor competes with UV radiation resistance associated gene product (UVRAG) for interaction with Beclin 1, and the complex formation of Barkor and Beclin1 is required for their localizations to autophagosomes. Therefore, we define a regulatory signaling pathway mediated by Barkor that positively controls autophagy through Beclin 1 and represents a potential target for drug development in the treatment of human diseases implicated in autophagic dysfunction.
Concentrations of heavy metals (Cd, Pb, Cu, Zn, and Ni) were measured in the foodstuffs, house dust, underground/drinking water, and soil from an electronic waste (e-waste) area in South China. ...Elevated concentrations of these potentially toxic metals were observed in the samples but not in drinking water. The health risks for metal exposure via food consumption, dust ingestion, and drinking water were evaluated for local residents. For the average residents in the e-waste area, the non-carcinogenic risks arise predominantly from rice (hazard index=3.3), vegetables (2.2), and house dust (1.9) for adults, while the risks for young children are dominated by house dust (15). Drinking water may provide a negligible contribution to risk. However, local residents who use groundwater as a water supply source are at high non-carcinogenic risk. The potential cancer risks from oral intake of Pb are 8×10−5 and 3×10−4 for average adults and children, and thus groundwater would have a great potential to induce cancer (5×10−4 and 1×10−3) in a highly exposed population. The results also reveal that the risk from oral exposure is much higher than the risk from inhalation and dermal contact with house dust.
•Elevated concentrations of toxic metals were observed but not in drinking water.•The risks for young children arise primarily from house dust.•Rice, vegetables, and house dust make the largest contribution to risk.•Drinking water may provide a negligible contribution to risk.•Groundwater has a great potential to induce cancer for some residents.
Combustion of domestic solid fuels is a significant source of polycyclic aromatic hydrocarbons (PAHs). Some oxygenated PAHs (o-PAHs) and PAHs with molecular weight of 302 (MW302 PAHs) are more toxic ...than the traditional 16 priority PAHs, whereas their emissions were much less elucidated. This study characterized the size-dependent emissions of parent PAHs (p-PAHs), o-PAHs, and MW302 PAHs from various combustion sources. The estimated emission factors (eEFs) from biomass burning sources were highest for most of the PAHs (391–8928 μg/kg), much higher than that of anthracite coal combustion (43.0–145 μg/kg), both which were operated in an indoor stove. Cigarette smoking had a high eEF of o-PAHs (240 ng/g). MW302 PAHs were not found in the emissions of smoking, cooking, and vehicular exhausts. Particle-size distributions of PAHs were compound- and source-dependent, and the tendency to associate with smaller particles was observed especially in biomass burning and cigarette smoking sources. Furthermore, the inter-source differences in PAH eEFs were associated with their dominance in fine particles. PAH composition profiles also varied with the particle size, showing increasing contributions of large-molecule PAHs with decreasing sizes in most cases. The size distributions of p-PAHs are much more significantly dependent on their n-octanol/air partition coefficients and vapor pressures than those of o-PAHs, suggesting differences in mechanisms governing their distributions. Several molecular diagnostic ratios (MDRs), including two based on MW302 PAHs, specific to these combustion scenarios were identified. However, the MDRs within some sources are also strongly size-dependent, providing a new explanation for the uncertainty in their application for source identification of PAHs. This work also highlights the necessity for understanding the size-resolved atmospheric behaviors and fate of PAHs after their emission.
•High EFs of PAHs from biomass burning and o-PAHs from cigarette smoking are found.•The particle-size distributions of PAHs are compound- and source-dependent.•PAH compositions from various biomass burning sources were generally similar.•PAH particle-size distributions are governed by their KOA and vapor pressure.•The molecular diagnostic ratios from combustion are also particle size-dependent.
In this paper, the graphic representation method is used to study the multiple characteristics of heart sounds from a resting state to a state of motion based on single- and four-channel heart-sound ...signals. Based on the concept of integration, we explore the representation method of heart sound and blood pressure during motion. To develop a single- and four-channel heart-sound collector, we propose new concepts such as a sound-direction vector of heart sound, a motion-response curve of heart sound, the difference value, and a state-change-trend diagram. Based on the acoustic principle, the reasons for the differences between multiple-channel heart-sound signals are analyzed. Through a comparative analysis of four-channel motion and resting-heart sounds, from a resting state to a state of motion, the maximum and minimum similarity distances in the corresponding state-change-trend graphs were found to be 0.0038 and 0.0006, respectively. In addition, we provide several characteristic parameters that are both sensitive (such as heart sound amplitude, blood pressure, systolic duration, and diastolic duration) and insensitive (such as sound-direction vector, state-change-trend diagram, and difference value) to motion, thus providing a new technique for the diverse analysis of heart sounds in motion.
Short-chain and medium-chain chlorinated paraffins (SCCPs and MCCPs) were measured in sediments from ponds, rivers and tributaries, and an estuary, as well as a sediment core in the Pearl River Delta ...(PRD), South China, to comprehensively investigate the spatial and temporal distributions of CPs. The concentrations of SCCPs and MCCPs in sediment were varied from 320 to 6600 ng/g and from 880 to 38 000 ng/g, respectively. Elevated CP concentrations were found in pond sediments (means of 2800 and 21 000 ng/g for SCCPs and MCCPs) in the e-waste recycling area and in river sediments (means of 1200 and 3900 ng/g for SCCPs and MCCPs) in the highly industrialized areas. The significant positive correlations between SCCP concentration and MCCPs/SCCPs in the highly industrialized areas reflected the emission of local industry activities, while the significant negative correlations in the low industrial activity areas could be linked to long-range transportation of CPs. An increased abundance of short chain and low chlorinated congeners was observed in the low industrial activity areas compared to the industrialized areas. The preferred transportation of short chain and low chlorinated congener CPs and the dechlorination of higher chlorinated congeners CPs were the most likely reasons. The vertical profile of CPs in the sediment core indicated a rapid increase in the usage of CPs and a shift to more MCCPs in recent years. The decreased chlorine content of CPs with increasing sediment depth indicated the possibility of dechlorination of higher chlorinated congeners (Cl9 and Cl10) after deposition in sediments with greater dechlorination potential for short chain CPs than long chain CPs.
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