Proteasome granular localization is regulated through mitochondrial respiration and kinase signaling

In yeast, proteasomes are enriched in cell nuclei where they execute important cellular functions. Nutrient-stress can change this localization indicating proteasomes respond to the cell's metabolic state. However, the signals that connect these processes remain poorly understood. Carbon starvation triggers a reversible translocation of proteasomes to cytosolic condensates known as proteasome storage granules (PSGs). Surprisingly, we observed strongly reduced PSG levels when cells had active cellular respiration prior to starvation. This suggests the mitochondrial activity of cells is a determining factor in the response of proteasomes to carbon starvation. Consistent with this, upon inhibition of mitochondrial function we observed proteasomes relocalize to granules. These links between proteasomes and metabolism involve specific signaling pathways, as we identified a MAP kinase cascade that is critical to the formation of proteasome granules after respiratory growth but not following glycolytic growth. Furthermore, the yeast homolog of AMP kinase, Snf1, is important for proteasome granule formation induced by mitochondrial inhibitors, while dispensable for granule formation following carbon starvation. We propose a model where mitochondrial activity promotes proteasome nuclear localization.

Dynamics of Energy Metabolism in Carbon Starvation-Induced Fruitlet Abscission in Litchi

Fruit abscission is triggered by multiple changes in endogenous components of the fruit, including energy metabolism. However, it is still unknown how the core energy metabolism pathways are modified during fruit abscission. Here, we investigated the relationship between carbon starvation-induced fruitlet abscission and energy metabolism changes in litchi. The fruitlet abscission of litchi ‘Feizixiao’ was induced sharply by girdling plus defoliation (GPD), a carbon stress treatment. Using liquid chromatography tandem mass spectrometry (LC-MS/MS) targeted metabolomics analysis, we identified a total of 21 metabolites involved in glycolysis, TCA cycle and oxidative phosphorylation pathways. Among them, the content of most metabolites in glycolysis pathways and TCA cycles was reduced, and the activity of corresponding metabolic enzymes such as ATP-dependent phosphofructokinase (ATP-PFK), pyruvate kinase (PK), citrate synthase (CS), succinate thiokinase (SAT), and NAD-dependent malate dehydrogenase (NAD-MDH) was decreased. Consistently, we further showed that the expression of the relative genes (LcPFK2, LcPK2, LcPK4, LcCS1, LcCS2, LcSAT, LcMDH1 and LcMDH2) was also significantly down-regulated. In contrast, the level of ATP, an important metabolite in the oxidative phosphorylation pathway, was elevated in parallel with both higher activity of H+-ATPase and the increased expression level of LcH+-ATPase1. In conclusion, our findings suggest that carbon starvation can induce fruitlet abscission in litchi probably by energy depletion that mediated through both the suppression of the glycolysis pathway and TCA cycle and the enhancement of the oxidative phosphorylation pathway.

Tree Mortality Risks Under Climate Change in Europe: Assessment of Silviculture Practices and Genetic Conservation Networks

General Context: Climate change can positively or negatively affect abiotic and biotic drivers of tree mortality. Process-based models integrating these climatic effects are only seldom used at species distribution scale.Objective: The main objective of this study was to investigate the multi-causal mortality risk of five major European forest tree species across their distribution range from an ecophysiological perspective, to quantify the impact of forest management practices on this risk and to identify threats on the genetic conservation network.Methods: We used the process-based ecophysiological model CASTANEA to simulate the mortality risk of Fagus sylvatica, Quercus petraea, Pinus sylvestris, Pinus pinaster, and Picea abies under current and future climate conditions, while considering local silviculture practices. The mortality risk was assessed by a composite risk index (CRIM) integrating the risks of carbon starvation, hydraulic failure and frost damage. We took into account extreme climatic events with the CRIMmax, computed as the maximum annual value of the CRIM.Results: The physiological processes' contributions to CRIM differed among species: it was mainly driven by hydraulic failure for P. sylvestris and Q. petraea, by frost damage for P. abies, by carbon starvation for P. pinaster, and by a combination of hydraulic failure and frost damage for F. sylvatica. Under future climate, projections showed an increase of CRIM for P. pinaster but a decrease for P. abies, Q. petraea, and F. sylvatica, and little variation for P. sylvestris. Under the harshest future climatic scenario, forest management decreased the mean CRIM of P. sylvestris, increased it for P. abies and P. pinaster and had no major impact for the two broadleaved species. By the year 2100, 38–90% of the European network of gene conservation units are at extinction risk (CRIMmax=1), depending on the species.Conclusions: Using a process-based ecophysiological model allowed us to disentangle the multiple drivers of tree mortality under current and future climates. Taking into account the positive effect of increased CO2 on fertilization and water use efficiency, average mortality risk may increase or decrease in the future depending on species and sites. However, under extreme climatic events, our process-based projections are as pessimistic as those obtained using bioclimatic niche models.

An l-2-hydroxyglutarate biosensor based on specific transcriptional regulator LhgR

Abstractl-2-Hydroxyglutarate (l-2-HG) plays important roles in diverse physiological processes, such as carbon starvation response, tumorigenesis, and hypoxic adaptation. Despite its importance and intensively studied metabolism, regulation of l-2-HG metabolism remains poorly understood and none of regulator specifically responded to l-2-HG has been identified. Based on bacterial genomic neighborhood analysis of the gene encoding l-2-HG oxidase (LhgO), LhgR, which represses the transcription of lhgO in Pseudomonas putida W619, is identified in this study. LhgR is demonstrated to recognize l-2-HG as its specific effector molecule, and this allosteric transcription factor is then used as a biorecognition element to construct an l-2-HG-sensing FRET sensor. The l-2-HG sensor is able to conveniently monitor the concentrations of l-2-HG in various biological samples. In addition to bacterial l-2-HG generation during carbon starvation, biological function of the l-2-HG dehydrogenase and hypoxia induced l-2-HG accumulation are also revealed by using the l-2-HG sensor in human cells.

Diversity in Starvation Survival Strategies and Outcomes among Heterotrophic Proteobacteria

Heterotrophic Proteobacteria are versatile opportunists that have been extensively studied as model organisms in the laboratory, as both pathogens and beneficial symbionts of plants and animals, and as ubiquitous organisms found free-living in many environments. Succeeding in these niches requires an ability to persist for potentially long periods of time in growth-arrested states when essential nutrients become limiting. The tendency of these bacteria to grow in dense biofilm communities frequently leads to the development of steep nutrient gradients and deprivation of interior cells even when the environment is nutrient rich. Surviving within host environments also likely requires tolerating growth arrest due to the host limiting access to nutrients and transitioning between hosts may require a period of survival in a nutrient-poor environment. Interventions to maximise plant-beneficial activities and minimise infections by bacteria will require a better understanding of metabolic and regulatory networks that contribute to starvation survival, and how these networks function in diverse organisms. Here we focus on carbon starvation as a growth-arresting condition that limits availability not only of substrates for biosynthesis but also of energy for ongoing maintenance of the electrochemical gradient across the cell envelope and cellular integrity. We first review models for studying bacterial starvation and known strategies that contribute to starvation survival<i>.</i> We then present the results of a survey of carbon starvation survival strategies and outcomes in ten bacterial strains, including representatives from the orders Enterobacterales and Pseudomonadales (both Gammaproteobacteria) and Burkholderiales (Betaproteobacteria). Finally, we examine differences in gene content between the highest and lowest survivors to identify metabolic and regulatory adaptations that may contribute to differences in starvation survival.

Pure-culture and mixed community biofilm responses to carbon-starvation and UV-C exposure /

Biofilms are known to contribute to disease through inherent protective mechanisms and propagation strategies. These multi-cellular systems also play essential roles in numerous environmental processes. The current study investigated the responses of a mixed community biofilm to carbon-starvation, and measured the effects of UV-C on pure-culture biofilms at different stages of maturity by monitoring metabolic and cell yield responses. Carbon dioxide production and biofilm-derived planktonic cell yield were used at the measurement parameters. The mixed community rapidly responded to induced carbon-starvation under continuous flow conditions by remaining metabolically inactive throughout the 96 and 120 h starvation periods, only to promptly return to a metabolically active state upon the reintroduction of carbon. The effects of UV-C on pure-culture biofilms was negligible, with no log activation being achieved, and metabolic activity remaining static.

Pure-culture and mixed community biofilm responses to carbon-starvation and UV-C exposure /

Biofilms are known to contribute to disease through inherent protective mechanisms and propagation strategies. These multi-cellular systems also play essential roles in numerous environmental processes. The current study investigated the responses of a mixed community biofilm to carbon-starvation, and measured the effects of UV-C on pure-culture biofilms at different stages of maturity by monitoring metabolic and cell yield responses. Carbon dioxide production and biofilm-derived planktonic cell yield were used at the measurement parameters. The mixed community rapidly responded to induced carbon-starvation under continuous flow conditions by remaining metabolically inactive throughout the 96 and 120 h starvation periods, only to promptly return to a metabolically active state upon the reintroduction of carbon. The effects of UV-C on pure-culture biofilms was negligible, with no log activation being achieved, and metabolic activity remaining static.