Survival In Adverse Conditions Research Articles

Contribution of Toxin-Antitoxin Systems to Adherent-Invasive E. coli Pathogenesis.

Pathobionts have been implicated in various chronic diseases, including Crohn's disease (CD), a multifactorial chronic inflammatory condition that primarily affects the gastrointestinal tract, causing inflammation and damage to the digestive system. While the exact cause of CD remains unclear, adherent-invasive Escherichia coli (AIEC) strains have emerged as key contributors to its pathogenesis. AIEC are characterized by their ability to adhere to and invade intestinal epithelial cells and survive and replicate inside macrophages. However, the mechanisms underlying the virulence and persistence of AIEC within their host remain the subject of intensive research. Toxin-antitoxin systems (TAs) play a potential role in AIEC pathogenesis and may be therapeutic targets. These systems generally consist of two components: a toxin harmful to the cell and an antitoxin that neutralizes the toxin's effects. They contribute to bacterial survival in adverse conditions and regulate bacterial growth and behavior, affecting various cellular processes in bacterial pathogens. This review focuses on the current information available to determine the roles of TAs in the pathogenicity of AIEC. Their contribution to the AIEC stress response, biofilm formation, phage inhibition, the maintenance of mobile genetic elements, and host lifestyles is discussed.

Unveiling the persistent threat: recent insights into Listeria monocytogenes adaptation, biofilm formation, and pathogenicity in foodborne infections.

Listeriosis is a severe disease caused by the foodborne pathogen Listeria monocytogenes, posing a significant risk to vulnerable populations such as the elderly, pregnant women, and newborns. While relatively uncommon, it has a high global mortality rate of 20-30%. Recent research indicates that smaller outbreaks of the more severe, invasive form of the disease occur more frequently than previously thought, despite the overall stable infection rates of L. monocytogenes over the past 10years. The ability of L. monocytogenes to form biofilm structures on various surfaces in food production environments contributes to its persistence and challenges in eradication, potentially leading to contamination of food and food production facilities. To address these concerns, this review focuses on recent developments in epidemiology, risk evaluations, and molecular mechanisms of L. monocytogenes survival in adverse conditions and environmental adaptation. Additionally, it covers new insights into strain variability, pathogenicity, mutations, and host vulnerability, emphasizing the important events framework that elucidates the biochemical pathways from ingestion to infection. Understanding the adaptation approaches of L. monocytogenes to environmental stress factors is crucial for the development of effective and affordable pathogen control techniques in the food industry, ensuring the safety of food production.

The longevity and reversibility of quiescence in Schizosaccharomyces pombe are dependent upon the HIRA histone chaperone

ABSTRACT Quiescence (G0) is a reversible non-dividing state that facilitates cellular survival in adverse conditions. Here, we demonstrate that the HIRA histone chaperone complex is required for the reversibility and longevity of nitrogen starvation-induced quiescence in Schizosaccharomyces pombe. The HIRA protein, Hip1 is not required for entry into G0 or the induction of autophagy. Although hip1Δ cells retain metabolic activity in G0, they rapidly lose the ability to resume proliferation. After a short period in G0 (1 day), hip1Δ mutants can resume cell growth in response to the restoration of a nitrogen source but do not efficiently reenter the vegetative cell cycle. This correlates with a failure to induce the expression of MBF transcription factor-dependent genes that are critical for S phase. In addition, hip1Δ G0 cells rapidly progress to a senescent state in which they can no longer re-initiate growth following nitrogen source restoration. Analysis of a conditional hip1 allele is consistent with these findings and indicates that HIRA is required for efficient exit from quiescence and prevents an irreversible cell cycle arrest.

Photosynthetic apparatus variability of the rigid hornwort (Ceratophyllum demersum L.) from the Saksagan River influenced by the mine activity

Aquatic ecosystems are the final absorbers of various pollutants, so require complex monitoring to control their condition. Macrophytes and other aquatic plants serve as reliable indicators of the hydro ecosystems ecological state, as they provide an integral assessment of the aquatic environment in time and space. Changes in chlorophyll a and chlorophyll b content and ratio in the rigid hornwort (Ceratophyllum demersum L.) leaves depending on the water hydro chemical parameters of Saksagan River in the zones of influence of "Ternivska" and "Oktyabrska" mines (Kryvyi Rih city) were studied. The purpose of the work was to find out the features of C. demersum photosynthetic apparatus reorganization as plants adaptive reaction to the technogenic transformation of river ecosystem. The water mineralization changes in the range of 1.2–2.9 g/L and fluctuations in the nitrates and nitrites content in the Saksagan River different sections were established. Both an increase and a decrease in the content of chlorophyll both forms in C. demersum leaves were detected, but more significant changes were related to chlorophyll b content. The photosynthetic pigments total content in plant leaves was the highest in the most mineralized areas (points of return water discharge and 500 m below of "Ternivska" mine), and it was the lowest due to sharp water mineralization increasing (500 m below the return water discharge of "Oktyabrska" mine). Significant variability in the photosynthetic pigments content and ratio indicates flexible regulation of their synthesis in C. demersum leaves to maintain the efficiency of the photosynthetic apparatus under conditions of water pollution. The revealed regularities indicate the active protection of the photosynthetic process in the C. demersum leaves, which is an important prerequisite for the plant’s survival in adverse conditions and the preservation of the structure and productivity of the water ecosystem of the Saksagan River.

Prolonged Subculturing of Aspergillus fumigatus on Galleria Extract Agar Results in Altered Virulence and Sensitivity to Antifungal Agents.

Aspergillus fumigatus is an environmental saprophyte and opportunistic fungal pathogen of humans. The aim of the work presented here was to examine the effect of serially subculturing A. fumigatus on agar generated from Galleria mellonella larvae in order to characterize the alterations in the phenotypes that might occur. The passaged strains showed alterations in virulence, antifungal susceptibility, and in protein abundances that may indicate adaptation after 25 passages over 231 days on Galleria extract agar. Passaged strains demonstrated reduced virulence in G. mellonella larvae and increased tolerance to hemocyte-mediated killing, hydrogen peroxide, itraconazole, and amphotericin B. A label-free proteomic analysis of control and passaged A. fumigatus strains revealed a total of 3329 proteins, of which 1902 remained following filtration, and 32 proteins were statistically significant as well as differentially abundant. Proteins involved in the response to oxidative stress were altered in abundance in the passaged strain and included (S)-S-oxide reductase (+2.63-fold), developmental regulator FlbA (+2.27-fold), and histone H2A.Z (-1.82-fold). These results indicate that the prolonged subculturing of A. fumigatus on Galleria extract agar results in alterations in the susceptibility to antifungal agents and in the abundance of proteins associated with the oxidative stress response. The phenomenon may be a result of selection for survival in adverse conditions and highlight how A. fumigatus may adapt to tolerate the pulmonary immune response in cases of human infection.

Absence of Wee1 alters global transcriptional response to oxidative stress in Schizosaccharomyces pombe.

Stress response and checkpoint activation are the main determinants of cellular survival in adverse conditions. In Schizosaccharomyces pombe, these are controlled by the Mitogen Activated Protein Kinase Spc1 and the Cyclin dependent Kinase Cdc2 respectively. Cdc2 is regulated positively by Cdc25 and negatively by Wee1. Changes in Cdc2 activity can be sensed by Spc1 resulting in the modulation of mitotic timing by Spc1. Functional cross talks between cell cycle regulation and MAPK machinery during regulation of mitotic timing are well characterised but the presence of similar communication during stress response remains unexplored. In this study we report how the checkpoint activator kinase Wee1 can also influence the transcriptional response to oxidative stress. We show that deletion of Wee1 results in changes in gene expression of the cells, especially with respect to genes whose expression is known to be regulated by Spc1. These differences are seen in unperturbed cells as well as during oxidative stress. Moreover, such variations extend beyond what could be expected to occur due to the known enhanced Spc1 activity of these cells. This is the first depiction of the influence of Wee1 and consequently Cdc2 activity on transcriptional response to oxidative stress.

Impaired Metabolic Plasticity Under Stress Constitutes a Therapeutic Vulnerability in IDH1/2-Mutant Acute Myeloid Leukemia

The metabolic state of leukemic cells can be defined by driver mutations and determine susceptibility to therapy. Metabolic rewiring is an essential feature of cancer cells to promote survival in adverse conditions and can be exploited as a therapeutic vulnerability. We previously showed metabolic adaptation upon PDK1 or complex II inhibition in FLT3-ITD Acute Myeloid Leukemias (AML) (Erdem et al, 2022). Mutations in Isocitrate Dehydrogenase (IDH1/2), an enzyme responsible for converting isocitrate into α-ketoglutarate (αKG), are found in about 20% of AML cases. These mutations can also impact on cellular metabolism due to their role in the tricarboxylic acid (TCA) cycle and abnormal production of 2-R-hydroxyglutarate, a competitive inhibitor of αKG-dependent enzymes, resulting in chromatin hypermethylation. However, the metabolic consequences of IDH mutations are still largely unknown. Here, we investigated the metabolic state of IDH1/2 mutant cells with the ultimate aim to uncover targetable vulnerabilities. To explore potential metabolic targets associated with IDH2 mutations, we performed RNA sequencing in CRISPR/Cas9 gene-edited isogenic TF1wt and IDH2R140Q cells. The top differently expressed genes were associated with fatty acid (FA) oxidation (FAO), such as CD36, a major FA transporter, and ACSL1, responsible for converting FAs into fatty-acyl-CoAs. Gene Set Enrichment Analysis associated the transcriptome of IDH2R140Q cells with processes related to increased mitochondrial complex I activity and FA biosynthesis. To functionally validate our findings, we performed a drug screen targeting the main metabolic pathways. Metformin, an FDA-approved drug known to inhibit complex I, selectively induced apoptosis in IDH2R140Q cells. Treatment with triacsin C, an ACSL1 inhibitor, reduced cell proliferation in the mutant cells, whereas 2-deoxyglucose, a glycolysis inhibitor, had no significant effect in this population. In contrast, TF1wt largely depended on glycolysis, indicating a metabolic rewiring after IDH2 mutation. Moreover, we confirmed by flow cytometry higher surface expression of CD36 in IDH2R140Q cells, which was downregulated upon treatment with FAO inhibitors and AG-221, an IDH2 inhibitor. Altogether, our data suggest that IDH2-mutant cells heavily rely on mitochondrial respiration, whereby FAs act as the main carbon source for this pathway. We then assessed oxygen consumption (OCR) and extracellular acidification (ECAR) rates by Seahorse analysis. In steady state, mitochondrial respiration was similar between TF1wt and IDH2R140Q cells, and blocking complex I with metformin significantly reduced OCR in both cell lines. However, its effect on cell viability was much more pronounced in IDH2R140Q cells. Interestingly, WT cellsshowed increased basal ECAR after treatment, suggestive of a shift towards glycolysis, unobserved in the mutant cells. We measured glucose consumption and lactate production in the cellular media, which confirmed a higher glucose uptake and lactate secretion only in the WT setting. Gene expression analysis revealed up-regulation of the glycolysis-related genes ALDO and PFKFB3 in WT but not IDH2R140Q cells upon treatment. Altogether, these data further confirm a metabolic rewiring of IDH2R140Q cells that now depend less on glycolysis to drive mitochondrial respiration. Next, we confirmed our findings in primary AML samples harboring IDH1/2-mutations. Treatment with metformin followed by Seahorse showed similar results as observed in theTF1 models. We previously developed a technology based on differential membrane protein expression that allowed us to identify and prospectively isolate NPM1cyt/IDH1R132H from NPM1cyt/IDH1wt subclones within an individual patient for functional studies (de Boer et al, 2018). RNA-seq analysis revealed distinct transcriptional signatures whereby the IDH1R132H clone was associated with processes related to mitochondrial and FA metabolism, and displayed a 3-fold higher expression of CD36 in comparison to the IDH1wt clone. While most mutations in cancer often lead to metabolic flexibility, our findings suggest that IDH1/2-mutant AMLs induce metabolic rewiring, but are less prone to readapt under stress conditions. Consequently, we uncover the lack of metabolic plasticity as the mechanism behind the usage of complex I inhibition as therapeutic vulnerability in IDH1/2-mutant AMLs. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Stress response modulation: the key to survival of pathogenic and spoilage bacteria during poultry processing.

The control of bacterial contaminants on meat is a key area of interest in the food industry. Bacteria are exposed to a variety of stresses during broiler processing which challenge bacterial structures and metabolic pathways causing death or sublethal injury. To counter these stresses, bacteria possess robust response systems that can induce shifts in the transcriptome and proteome to enable survival. Effective adaptive responses, such as biofilm formation, shock protein production and metabolic flexibility, require rapid induction and implementation at a cellular and community level to facilitate bacterial survival in adverse conditions. This review aims to provide an overview of the scientific literature pertaining to the regulation of complex adaptive processes used by bacteria to survive the processing environment, with particular focus on species that impact the quality and safety of poultry products like Campylobacter spp., Salmonella enterica and Pseudomonas spp.

The metabolic depression and revival of purple urchins (Strongylocentrotus purpuratus) in response to macroalgal availability

Organisms can reduce their metabolism and/or decrease energy allocation to reproduction in order to survive starvation during extended periods of food limitation. This is especially important for coastal rocky reefs where widespread kelp deforestation has become increasingly common in recent decades. This deforestation often results in the formation of urchin barrens that have high densities of herbivorous sea urchins and little macroalgae for them to consume. While it is known that these barrens can persist for years to decades, and it has been suggested that urchins can reduce their metabolisms and/or consume microalgae during these periods, the mechanisms that allow for such prolonged survival in adverse conditions, and if these are mechanisms are reversible are not entirely understood. Here, we provide experimental evidence to show that urchin metabolism and gonad mass both decrease significantly when the urchins are deprived of macroalgae, and that these urchins regain normal metabolic activity and gonad masses when access to this food resource is restored. Our results have important implications for consumers in other ecosystems where access to energetic resources is spatially or temporally variable and can point to new avenues of research to explain how organisms adjust their energetic needs to survive extended periods of starvation.

Molecular Regulation of Paused Pluripotency in Early Mammalian Embryos and Stem Cells.

The energetically costly mammalian investment in gestation and lactation requires plentiful nutritional sources and thus links the environmental conditions to reproductive success. Flexibility in adjusting developmental timing enhances chances of survival in adverse conditions. Over 130 mammalian species can reversibly pause early embryonic development by switching to a near dormant state that can be sustained for months, a phenomenon called embryonic diapause. Lineage-specific cells are retained during diapause, and they proliferate and differentiate upon activation. Studying diapause thus reveals principles of pluripotency and dormancy and is not only relevant for development, but also for regeneration and cancer. In this review, we focus on the molecular regulation of diapause in early mammalian embryos and relate it to maintenance of potency in stem cells in vitro. Diapause is established and maintained by active rewiring of the embryonic metabolome, epigenome, and gene expression in communication with maternal tissues. Herein, we particularly discuss factors required at distinct stages of diapause to induce, maintain, and terminate dormancy.

RND Efflux Systems Contribute to Resistance and Virulence of Aliarcobacter butzleri.

Aliarcobacter butzleri is an emergent enteropathogen that can be found in a range of environments. This bacterium presents a vast repertoire of efflux pumps, such as the ones belonging to the resistance nodulation cell division family, which may be associated with bacterial resistance, as well as virulence. Thus, this work aimed to evaluate the contribution of three RND efflux systems, AreABC, AreDEF and AreGHI, in the resistance and virulence of A. butzleri. Mutant strains were constructed by inactivation of the gene that encodes the inner membrane protein of these systems. The bacterial resistance profile of parental and mutant strains to several antimicrobials was assessed, as was the intracellular accumulation of the ethidium bromide dye. Regarding bacterial virulence, the role of these three efflux pumps on growth, strain fitness, motility, biofilm formation ability, survival in adverse conditions (oxidative stress and bile salts) and human serum and in vitro adhesion and invasion to Caco-2 cells was evaluated. We observed that the mutants from the three efflux pumps were more susceptible to several classes of antimicrobials than the parental strain and presented an increase in the accumulation of ethidium bromide, indicating a potential role of the efflux pumps in the extrusion of antimicrobials. The mutant strains had no bacterial growth defects; nonetheless, they presented a reduction in relative fitness. For the three mutants, an increase in the susceptibility to oxidative stress was observed, while only the mutant for AreGHI efflux pump showed a relevant role in bile stress survival. All the mutant strains showed an impairment in biofilm formation ability, were more susceptible to human serum and were less adherent to intestinal epithelial cells. Overall, the results support the contribution of the efflux pumps AreABC, AreDEF and AreGHI of A. butzleri to antimicrobial resistance, as well as to bacterial virulence.

Esporas de inclusión

La educación inclusiva al ser concebida en tanto espora emerge con fines de dispersión y supervivencia en condiciones adversas. El nomadismo epistemológico de la educación inclusiva inaugura una nueva forma de concebir y practicar el trabajo heurístico. El nomadismo epistemológico de la educación inclusiva es una forma analítica dinámica, un espacio de intensos flujos, una red de apertura, creación y relación de diversas singularidades y recursos epistemológicos –principio de heterogénesis–. La naturaleza del dominio reafirma un carácter transposicional, fundamentalmente, articulado a partir de complejos y singulares flujos de movilidad y referencias cruzadas entre (inter)disciplinas, teorías, conceptos, niveles discursivos, etc. Es un territorio que se desplaza por diversos lugares produciendo-a-sí-mismo y creando sus propios efectos específicos. El trabajo describe ocho claves analíticas cruciales en la comprensión epistémica de su dominio.

Biological functions of the autophagy-related proteins Atg4 and Atg8 in Cryptococcus neoformans.

Autophagy is a mechanism responsible for intracellular degradation and recycling of macromolecules and organelles, essential for cell survival in adverse conditions. More than 40 autophagy-related (ATG) genes have been identified and characterized in fungi, among them ATG4 and ATG8. ATG4 encodes a cysteine protease (Atg4) that plays an important role in autophagy by initially processing Atg8 at its C-terminus region. Atg8 is a ubiquitin-like protein essential for the synthesis of the double-layer membrane that constitutes the autophagosome vesicle, responsible for delivering the cargo from the cytoplasm to the vacuole lumen. The contributions of Atg-related proteins in the pathogenic yeast in the genus Cryptococcus remain to be explored, to elucidate the molecular basis of the autophagy pathway. In this context, we aimed to investigate the role of autophagy-related proteins 4 and 8 (Atg4 and Atg8) during autophagy induction and their contribution with non-autophagic events in C. neoformans. We found that Atg4 and Atg8 are conserved proteins and that they interact physically with each other. ATG gene deletions resulted in cells sensitive to nitrogen starvation. ATG4 gene disruption affects Atg8 degradation and its translocation to the vacuole lumen, after autophagy induction. Both atg4 and atg8 mutants are more resistant to oxidative stress, have an impaired growth in the presence of the cell wall-perturbing agent Congo Red, and are sensitive to the proteasome inhibitor bortezomib (BTZ). By that, we conclude that in C. neoformans the autophagy-related proteins Atg4 and Atg8 play an important role in the autophagy pathway; which are required for autophagy regulation, maintenance of amino acid levels and cell adaptation to stressful conditions.

Communication between Cyclin-dependent kinase Cdc2 and the Wis1-Spc1 MAPK pathway determines mitotic timing in Schizosaccharomyces pombe.

ABSTRACTCheckpoint activation and gene expression modulation represent key determinants of cellular survival in adverse conditions. The former is regulated by cyclin-dependent kinases (CDKs) while the latter can be controlled by mitogen-activated protein kinases (MAPKs). Association between cell-cycle progression and MAPK-dependent gene expression exists in cells growing in optimal environments. While MAPK-mediated regulation of the cell cycle is well characterised, the reciprocal influence of mitotic CDK on stress response is not well studied. We present evidence that CDK activity can regulate the extent of MAPK activation in Schizosaccharomyces pombe cells. We show that increasing or decreasing mitotic CDK (Cdc2) activity in S. pombe cells can affect the activation of stress responsive MAPK (Spc1) even in the absence of stress stimuli. Our results indicate that the strong correlation between Cdc2 activity and Spc1 MAPK-activity in S. pombe is important in regulating mitotic timing.This article has an associated First Person interview with the first author of the paper.

Assessing of adaptation ability of Listeria to benzalkonium chloride (BAC) by determination of its minimal inhibitory concentration

Bacteria of the genus Listeria pose a problem in the food industry due to their wide distribution and their good survival in adverse conditions. L. monocytogenes (E. Murray et al. 1926) Pirie 1940 is human pathogen, while L. innocua Seeliger (ATCC® 33090™) as not pathogenic bacteria is the most often found listeria in food production environment. Disinfectants represents an important part of Listeria management in food processing environments and benzalkonium chloride (BAC) is used frequently. The purpose of the work was to determine whether strains of listeria can adapt to BAC. To carry out the adaptation, a precise determination of antibacterial activity of BAC was needed. Firstly minimum inhibitory concentration (MICMTP) of BAC was determined with broth microdilution method for each Listeria strain. Then, we checked whether MICMTP was indeed the lowest concentration of BAC, which had an influence on growth of strains with growth curves. We found out that growth inhibitory effect (MICGC) was achieved at concentrations of BAC that were lower than MICMTP (0.1-0.5x of MICMTP values). Adaptation of listeria to BAC was therefore performed by using 0.25x MICGC as the initial BAC concentration. Results showed that 50 % of the strains were able to adapt to BAC, and in L. monocytogenes ŽM500 this adaptation was even stable. The broth microdilution method was useful for approximate assessment of antimicrobial activity of BAC, while for the more precise determination of disinfectant activity it is necessary to determine it by using another method such as plate count method.

When Three Isn't a Crowd: A Digyny Concept for Treatment-Resistant, Near-Triploid Human Cancers.

Near-triploid human tumors are frequently resistant to radio/chemotherapy through mechanisms that are unclear. We recently reported a tight association of male tumor triploidy with XXY karyotypes based on a meta-analysis of 15 tumor cohorts extracted from the Mitelman database. Here we provide a conceptual framework of the digyny-like origin of this karyotype based on the germline features of malignant tumors and adaptive capacity of digyny, which supports survival in adverse conditions. Studying how the recombinatorial reproduction via diploidy can be executed in primary cancer samples and HeLa cells after DNA damage, we report the first evidence that diploid and triploid cell sub-populations constitutively coexist and inter-change genomes via endoreduplicated polyploid cells generated through genotoxic challenge. We show that irradiated triploid HeLa cells can enter tripolar mitosis producing three diploid sub-subnuclei by segregation and pairwise fusions of whole genomes. Considering the upregulation of meiotic genes in tumors, we propose that the reconstructed diploid sub-cells can initiate pseudo-meiosis producing two “gametes” (diploid “maternal” and haploid “paternal”) followed by digynic-like reconstitution of a triploid stemline that returns to mitotic cycling. This process ensures tumor survival and growth by (1) DNA repair and genetic variation, (2) protection against recessive lethal mutations using the third genome.

Unraveling the role of the transcriptional regulator VirS in low pH–induced responses of Mycobacterium tuberculosis and identification of VirS inhibitors

The ability of Mycobacterium tuberculosis to respond and adapt to various stresses such as oxygen/nitrogen radicals and low pH inside macrophages is critical for the persistence of this human pathogen inside its host. We have previously shown that an AraC/XylS-type transcriptional regulator, VirS, which is induced in low pH, is involved in remodeling the architecture of the bacterial cell envelope. However, how VirS influences gene expression to coordinate these pH responses remains unclear. Here, using a genetic biosensor of cytoplasmic pH, we demonstrate that VirS is required for the intracellular pH maintenance in response to acidic stress and inside acidified macrophages. Furthermore, we observed that VirS plays an important role in blocking phagosomal-lysosomal fusions. Transcriptomics experiments revealed that VirS affects the expression of genes encoding metabolic enzymes, cell-wall envelope proteins, efflux pumps, ion transporters, detoxification enzymes, and transcriptional regulators expressed under low-pH stress. Employing electrophoretic mobility-shift assays, DNA footprinting, and in silico analysis, we identified a DNA sequence to which VirS binds and key residues in VirS required for its interaction with DNA. A significant role of VirS in M. tuberculosis survival in adverse conditions suggested it as a potential anti-mycobacterial drug target. To that end, we identified VirS inhibitors in a virtual screen; the top hit compounds inhibited its DNA-binding activity and also M. tuberculosis growth in vitro and inside macrophages. Our findings establish that VirS mediates M. tuberculosis responses to acidic stress and identify VirS-inhibiting compounds that may form the basis for developing more effective anti-mycobacterial agents.

LNK suppresses interferon signaling in melanoma

LNK (SH2B3) is a key negative regulator of JAK-STAT signaling which has been extensively studied in malignant hematopoietic diseases. We found that LNK is significantly elevated in cutaneous melanoma; this elevation is correlated with hyperactive signaling of the RAS-RAF-MEK pathway. Elevated LNK enhances cell growth and survival in adverse conditions. Forced expression of LNK inhibits signaling by interferon-STAT1 and suppresses interferon (IFN) induced cell cycle arrest and cell apoptosis. In contrast, silencing LNK expression by either shRNA or CRISPR-Cas9 potentiates the killing effect of IFN. The IFN-LNK signaling is tightly regulated by a negative feedback mechanism; melanoma cells exposed to IFN upregulate expression of LNK to prevent overactivation of this signaling pathway. Our study reveals an unappreciated function of LNK in melanoma and highlights the critical role of the IFN-STAT1-LNK signaling axis in this potentially devastating disease. LNK may be further explored as a potential therapeutic target for melanoma immunotherapy.

Life on the fringe: microbial adaptation to growth on carbon monoxide.

Microbial adaptation to extreme conditions takes many forms, including specialized metabolism which may be crucial to survival in adverse conditions. Here, we analyze the diversity and environmental importance of systems allowing microbial carbon monoxide (CO) metabolism. CO is a toxic gas that can poison most organisms because of its tight binding to metalloproteins. Microbial CO uptake was first noted by Kluyver and Schnellen in 1947, and since then many microbes using CO via oxidation have emerged. Many strains use molecular oxygen as the electron acceptor for aerobic oxidation of CO using Mo-containing CO oxidoreductase enzymes named CO dehydrogenase. Anaerobic carboxydotrophs oxidize CO using CooS enzymes that contain Ni/Fe catalytic centers and are unrelated to CO dehydrogenase. Though rare on Earth in free form, CO is an important intermediate compound in anaerobic carbon cycling, as it can be coupled to acetogenesis, methanogenesis, hydrogenogenesis, and metal reduction. Many microbial species—both bacteria and archaea—have been shown to use CO to conserve energy or fix cell carbon or both. Microbial CO formation is also very common. Carboxydotrophs thus glean energy and fix carbon from a “metabolic leftover” that is not consumed by, and is toxic to, most microorganisms. Surprisingly, many species are able to thrive under culture headspaces sometimes exceeding 1 atmosphere of CO. It appears that carboxydotrophs are adapted to provide a metabolic “currency exchange” system in microbial communities in which CO arising either abiotically or biogenically is converted to CO 2 and H 2 that feed major metabolic pathways for energy conservation or carbon fixation. Solventogenic CO metabolism has been exploited to construct very large gas fermentation plants converting CO-rich industrial flue emissions into biofuels and chemical feedstocks, creating renewable energy while mitigating global warming. The use of thermostable CO dehydrogenase enzymes to construct sensitive CO gas sensors is also in progress.

Nitrogen-Fixing Heterocystous Cyanobacteria in the Tonian Period

Cyanobacteria were the ultimate ancestor of all plastids and, for much of Earth's history, the only source of biogenic oxygen and a major source of fixed carbon and nitrogen. One cyanobacterial clade, subsections IV+V, is characterized by multicellularity and cell differentiation, with many members bearing specialized nitrogen-fixing (or diazotrophic) heterocysts and encysting akinetes [1-3]. Molecular clockestimates of the divergence time of this cladearehighly variable, ranging from ∼2,000 Ma (mega-annum) [4-9] to ∼500 Ma [10]. The older estimates are invariably calibrated by putative akinete fossilsfrom Paleoproterozoic-Mesoproterozoic rocks around 2,100-1,400 Ma [3, 11, 12]. However, the interpretation of these fossils as akinetes has been questioned [13], and the next oldest akinete and heterocyst fossils are ∼410 Ma [14]. Thus, the scarcity of reliable heterocystous cyanobacterial fossils significantly hampers our understanding of the evolution of complex multicellularity among cyanobacteria, their role in regulating geochemical cycles in the geological past, and our ability to calibrate cyanobacterial molecular clocks. Here, we report Tonian (∼1,000-720 Ma) filamentous cyanobacteria that are characterized by large cells, binary fission (for filament elongation), hormogonia (for asexual reproduction and dispersal), probable akinetes (for survival in adverse conditions), and by implication, diazotrophic heterocysts. The new fossils provide a minimum age calibration on the divergence of subsections IV+V and place a firm constraint on the evolution of akinetes and heterocysts.