Biology subjects

Westhoff, P.

Publications and source records attributed to Westhoff, P..

At least 19 recordsLinked to original sources

bioRxiv2026

The multidrug transporter Pdr5 from Saccharomyces cerevisiae is an ergosterol-modulated efflux pump

The pleiotropic drug resistance ABC transporter Pdr5 and its homologs play a critical role in multidrug resistance of numerous fungi, including pathogens. Previous studies demonstrated that alterations in the composition of the plasma membrane in which Pdr5 is embedded, specifically the ergosterol content, affected transport activity. Here we report the cryo-EM map of Saccharomyces cerevisiae Pdr5 in complex with ATP/ADP at 2.2 [A] resolution, revealing densities on the surface of the transmembrane domains as well as inside the substrate-binding site, that were assigned as ergosterol, identified and quantified by GC-MS, and the sphingolipid M(IP)2C, identified via native MS. The ergosterol molecules are bound to the protein in well-defined grooves and unexpected locations. We found that the activity of Pdr5 follows a Gaussian-like distribution, increasing with the ergosterol-concentration to reach a maximum before declining at higher concentrations, but this modulation was not observed in the presence of cholesterol, the sterol analogue found in mammalian cells. Mutational analysis corroborates the role of a single ergosterol-binding site in the modulation of Pdr5 activity. Our structural analysis identifies a protein-lipid network that could serve as a center for lipid engagement linking the surrounding lipid environment to the substrate-binding region of Pdr5. These findings advance our understanding of Pdr5 function by demonstrating that the lipid environment is closely linked to Pdr5 structure and activity.

biochemistry↗

bioRxiv2026

Lysine biosynthesis impairment shapes heat-stress acclimation through metabolic and transcriptional reprogramming in Arabidopsis thaliana

Global warming is increasing the frequency and intensity of high-temperature episodes, limiting plant productivity. However, the molecular mechanisms integrating primary metabolism with the heat stress response remains poorly understood. Here, we show that lysine biosynthesis contributes to the coordination of physiological, metabolic and transcriptional responses to heat stress in Arabidopsis thaliana. We compared wild-type, the lysine-biosynthesis mutant dapat, and the salicylic acid (SA)-biosynthesis and signaling mutants sid2-1 and npr1-3 under prolonged warming (6C above control for 7 days) and heat shock (38C for 6 h), followed by recovery. We assessed growth, gas exchange, photosynthetic performance, free SA, salicylic acid glucoside (SAG), salicylic acid glucose ester (SGE), and total SA content, primary metabolite profiles, heat-stress-responsive gene expression and transcriptome-wide changes by RNA sequencing. Before heat stress, dapat mutant presented a distinct metabolic state, marked by amino-acid accumulation, altered organic-acid profiles, reduced soluble sugars and elevated endogenous SA. This metabolic configuration persisted during prolonged warming, whereas WT and SA-pathway mutants underwent more dynamic reprogramming. Heat shock, by contrast, elicited a more convergent response across genotypes. Despite reduced basal PSII efficiency, dapat maintained photosynthetic performance during prolonged warming and recovered. Its transcriptional response, however, differed from that of WT and SA-pathway mutants: selected heat-responsive genes were constitutively or more strongly expressed, whereas some canonical heat-stress regulators showed weaker induction after heat shock. RNA-seq further revealed a largely conserved core heat-shock response but genotype-dependent regulation of defense, hormone and amino-acid-metabolism programs, particularly during recovery. Together, these findings indicate that impaired DAPAT activity establishes a metabolically primed but energetically constrained state that reshapes gas exchange, photosynthetic acclimation and heat-responsive transcription. Lysine homeostasis therefore emerges as a regulatory node linking primary metabolism and SA accumulation with SA-dependent and SA-independent components of heat-stress acclimation.

plant biology↗

bioRxiv2026

Root endophyte sulfur metabolites enhance redox balance and drought tolerance in Arabidopsis

Drought is a major consequence of climate change and significantly limits crop productivity. Plant growth-promoting bacteria offer a promising solution to mitigate drought stress. The root endophyte Pseudomonas argentinensis SA190 has been shown to enhance plant performance under drought stress conditions, but the mechanistic basis of SA190s beneficial effects remained unclear. Given the pivotal role of sulfur-containing compounds in abiotic stress responses, we investigated multiple sulfur-related Arabidopsis mutants under drought stress. We found that SA190 enhances sulfate uptake and promotes glutathione (GSH) accumulation in shoots under stress conditions. SA190 treatment improved the GSH/GSSG ratio, indicating an enhanced redox balance under drought. Selective inhibition of Arabidopsis GSH biosynthesis using buthionine sulfoximine (BSO) confirmed the essential contribution of bacterial GSH to drought stress. In addition, by generation and use of bacterial mutants deficient in the GSH synthesis pathway, we show that the bacteria directly provide Arabidopsis with either GSH or its precursor {gamma}-EC. In summary, SA190 promotes drought tolerance by supplying the host plant with additional GSH thereby maintaining cellular redox homeostasis and enhancing drought stress resilience.

Plant Biology↗

bioRxiv2026

Endosomal mRNA transport coordinates local mitochondrial bioenergetics during polar fungal growth

Mitochondrial function relies on the precise spatial coordination of protein synthesis and import. Most mitochondrial proteins are nuclear-encoded and must be supplied across varying intracellular distances. In highly polarized cells such as fungal hyphae and neurons, active long-distance mRNA transport is thought to sustain distal mitochondrial function, but its mechanistic coupling to protein import and organelle physiology is unclear. Here, we demonstrate that endosomal transport of mRNAs encoding mitochondrial proteins orchestrates local bioenergetics in infectious hyphae of Ustilago maydis. Using the subunit Atp3 of electron transport chain Complex V as a model, we uncover that the endosomal mRNA transporter Rrm4 is required for efficient mitochondrial protein import, particularly at growth poles. Loss of Rrm4 leads to defects in mitochondrial import, resulting in altered physiology. We propose that endosome-coupled mRNA transport constitutes a fundamental layer of subcellular mitochondrial homeostasis, with implications extending from fungal pathogenicity to neuronal disease. Significance StatementFungal pathogens depend on efficient polar growth to execute their infection programs. Consequently, their growing cell poles face a massive local demand of energy, which is supplied by mitochondria. Currently, it is unclear how involved proteins of the mitochondrial electron transport chain (ETC) reach these distant organelles. Here, we combine fungal genetics, metabolomics, transcriptomics and minimal invasive live-cell imaging to resolve this spatial challenge in the corn pathogen Ustilago maydis. We discover that long-distance endosomal hitchhiking of mRNAs encoding mitochondrial ETC components is essential to sustain active mitochondria at the expanding pole. Ultimately, this membrane-coupled mRNA trafficking precisely orchestrates subcellular mitochondrial function, disclosing a previously unrecognized Achilles heel for the development of novel fungicides.

microbiology↗

bioRxiv2026

Phylogeny, thermal seasonality, and life history shape trait variation in annual and perennial barley relatives

O_LIThere is growing interest in developing perennial cereal crops for sustainable production, yet key differences in trait syndromes between annual and perennial grasses and their influence on environmental adaptation remain poorly understood. C_LIO_LIWe measured 25 traits in 16 annual and perennial Hordeum species (45 accessions), including barley, grown over three seasons in a common garden. Using a phylogenetic framework and repeated transitions between annual and perennial forms, we (i) identified traits distinguishing these life strategies and (ii) tested how they relate to climate at the accessions origins. C_LIO_LIWild and cultivated barley are distinguished within the Hordeum clade by high growth rates and large organs, which may have predisposed wild barley to domestication. Annual and perennial accessions differed in resource allocation: annuals had higher harvest index and leaf and grain nitrogen, while perennials produced carbon-rich tissues and sustained vegetative growth. Seasonal temperature variation shaped trait syndromes: annual traits aligned with temperature in the driest quarter, reflecting selection under terminal stress, while perennial traits correlated with temperature in the wettest quarter, the main growth phase shaping long-term performance and survival. C_LIO_LIWe provide important information on traits and climate adaptations underlying perennial persistence and annual resource strategies, relevant for developing perennial cereal crops. C_LI

ecology↗

bioRxiv2026

Multi-omic data fusion reveals the in vivo enzyme kinetics of Vibrio natriegens at the genome-scale

Vibrio natriegens is a halophilic, Gram-negative marine bacterium that is increasingly used in metabolic engineering applications due to its fast growth rate. In sparse minimal medium the organism has a doubling time of 25 minutes, which is about twice as fast as Escherichia coli under similar conditions. Given that its protein density is similarly constrained to that of E. coli, this necessitates that its metabolic enzymes are able to catalyze flux at a higher rate to sustain its metabolism. In this work, we measure the apparent turnover numbers of metabolically active enzymes in V. natriegens under a variety of growth conditions. The apparent turnover numbers of V. natriegens enzymes were measured in vivo by conducting coupled quantitative proteomics and 13C metabolic flux analysis experiments under seven different carbon source conditions in sparse minimal medium. A high quality genome-scale metabolic model was constructed and curated using additional experimental data. This model was extended with enzyme constraints, and subsequently used to find kinetic parameters that minimize the difference between model predictions and experimental observations. This model guided data fusion approach enabled the estimation of 357 apparent turnover numbers for metabolically active enzymes in V. natriegens. Our results reveal that the metabolic enzymes of V. natriegens are in median 14-fold faster than those of E. coli under similar conditions. Moreover, we show that machine learning generated turnover number estimates substantially underestimate the kinetics of V. natriegens. Our turnover number estimates were used to parameterize multiple condition dependent enzyme constrained flux balance analysis models of V. natriegens, which improved their predictive accuracy compared to the machine learning parameterisation. The combined experimental-computational approach employed here sheds light on the mechanism V. natriegens uses to accelerate its growth. This approach can also be extended to other bacteria, increasing the availability of in vivo measured enzyme turnover numbers, and improving the predictive accuracy of enzyme constrained metabolic models of other microbes.

systems biology↗

bioRxiv2026

Context-dependent siderophore exploitability shapes microbial community structure

1)Siderophores are classically viewed as shared iron-scavenging public goods, yet their ecological roles in multispecies communities remain poorly defined. Here, we establish a synthetic microbial community to dissect how different siderophores, their uptake compatibility and spatial structure shape iron competition. Using Corynebacterium glutamicum as a model, we show that this siderophore non-producer accesses diverse xenosiderophores, including enterobactin secreted by Escherichia coli. However, exploitation was constrained and co-cultures converged to stable compositions. Dose-response experiments combined with mathematical modelling indicated that the producer retains more effective access to enterobactin than the exploiter. Presence of Pseudomonas putida altered this interaction, as it exploited enterobactin while producing pyoverdine, a siderophore inaccessible to the other community members that restricted their iron access. Across different cultivation scales, community dynamics was strongly influenced by spatial organization and initial composition. These findings identify siderophores as context-dependent iron-allocation agents that can promote microbial coexistence or exclusion.

microbiology↗

bioRxiv2026

Glycogen deficiency impairs diurnal energy metabolism and cell division in Synechocystis

Diurnal changes in light availability are a defining feature of life on Earth. Photoautotrophic organisms therefore store reduced carbon during the day to sustain energy metabolism at night. In cyanobacteria, glycogen is the primary carbon storage compound and supports both energy homeostasis and stress responses. Although glycogen-deficient Synechocystis strains have been studied previously, how these mutants cope with the loss of the major daytime carbon sink and can sustain themselves during the night remains unclear. Using single-cell microfluidics, transcriptomics, and metabolomics, we show that {Delta}glgC mutants exhibit pronounced light sensitivity. At sub-lethal light intensities, daytime transcriptional responses are dominated by downregulation of photosynthesis-related genes, likely preventing NADPH overaccumulation in the absence of a carbon sink. During the night, mutants display severe energy limitation, characterized by reduced ATP levels, altered redox balance, and depletion of central carbon intermediates. In contrast, fumarate and malate accumulate, indicating enhanced respiratory flux through succinate dehydrogenase. These metabolic constraints lead to extended lag phases and delayed cell divisions after the onset of light, demonstrating that glycogen-deficient cells fail to efficiently reinitiate growth after dawn. Overall, our results as a snapshot of the initial response to diurnal regimes highlight glycogen as a central integrator of diurnal physiology in Synechocystis, coordinating energy metabolism, redox balance, and cell division, with implications for metabolic robustness and the evolutionary constraints shaping (endo)symbiosis. Short summaryGlycogen deficiency disrupts day-night energy and redox homeostasis in Synechocystis, revealing constraints on growth, division, and symbiotic potential.

plant biology↗

bioRxiv2026

A hidden T-DNA-linked inversion-duplication causes a pronounced light-dependent phenotype in Arabidopsis

T-DNA insertion mutants are widely used to disrupt genes and infer their functions, yet the insertions can also trigger unintended genomic changes that confound phenotypic interpretation. Here, we used T-DNA insertion mutants affecting the major mitochondrial malate dehydrogenase (MDH1) and the heterodimeric NAD-dependent malic enzymes (ME1 and ME2) to examine their functional coordination across photoperiods and irradiance regimes. Under short days, especially at low light intensity, mdh1xme2 mutants were markedly smaller than wild type and, unexpectedly, than the mdh1xme1xme2 triple mutant, and they showed a more pronounced reduction in photosynthetic capacity. ME1 was undetectable in mdh1xme2, implying that the double and triple mutants effectively lack heterodimeric ME and should therefore behave similarly, contrary to what we observed. Whole-genome analysis resolved this discrepancy by revealing that the MDH1 T-DNA insertion in mdh1xme2 is accompanied by a major rearrangement, a 137-kbp duplication downstream of the insertion site, which was absent in the mdh1xme1xme2 triple mutant. This duplication increased gene dosage and elevated transcript abundance across the duplicated interval, while proteomics detected 5 of the 38 encoded proteins, including PEPC1. mdh1xme2 accumulated oxaloacetate-derived amino acids and displayed an altered carbon/nitrogen balance, making PEPC1 a plausible contributor to the exacerbated mdh1xme2 phenotype. Together, our data indicate that a T-DNA-linked structural variant can amplify expression of dozens of genes and intensify phenotypes at specific conditions, thereby affecting the interpretation of genotype-phenotype relationships. Because Agrobacterium-mediated DNA transfer also underpins many genome-editing workflows, our findings argue that structural validation around insertion/editing loci should be considered essential when interpreting T-DNA-derived plant lines.

plant biology↗

bioRxiv2026

Differential photosynthetic response to phosphate starvation in C3 and C4 Flaveria species

C4 photosynthesis is a CO2-concentration mechanism that separates CO2 fixation between two cell types, thereby reducing photorespiration and making C4 plants more efficient than their C3 counterparts. While the C4 cycle has evolved multiple times across different genera, this study evaluates very closely related C3 and C4 species within the genus Flaveria. Apart from their carbon metabolism, C4 plants also possess adaptations in their mineral nutrition. One key nutrient which is also directly involved in photosynthesis is phosphorus. It is absorbed by the plant in the form of inorganic phosphate and is an essential component of DNA, ATP, lipids, and carbohydrates. In the Flaveria C4 species, but not in the C3 species, phosphate limitation was shown to affect the dark reactions of photosynthesis. This study investigates how phosphate deficiency impacts the light reactions in C3 and C4 Flaveria plants. We observed a differential response in the functionality of photosynthetic energy conversion between the two species. When exposed to a limited phosphate supply, the C3 species reduced its linear electron transport rate while dissipating excess energy through high-energy quenching, which was regulated by a higher pH gradient across the thylakoid membrane. In contrast, the C4 species did not regulate its photosynthetic light reaction under phosphate limitation. Instead, it exhibited increased stress levels, evidenced by a stronger biomass reduction and the induction of stress markers in the leaves. Additionally, this study uncovered an acceleration in NPQ relaxation during phosphate limitation, regardless of the photosynthesis type. HighlightPhosphate deficiency reduced linear electron transport rates and induced dissipation of excess energy through non-photochemical quenching in the C3 Flaveria species, while in the C4 species, despite elevated stress levels, the photosynthetic light reactions were unaffected.

plant biology↗

bioRxiv2026

Mitochondrial malate metabolism acts as a control hub for photosynthesis and carbon-nitrogen balance in Arabidopsis

Malate is a central metabolite in plant energy metabolism and biosynthesis and serves as a major carrier of carbon and reducing equivalents between chloroplasts, the cytosol, and mitochondria. However, how individual malate-converting systems contribute to physiology in specific subcellular compartments remains incompletely understood. Here, we investigated the impact of combined loss of mitochondrial malate dehydrogenase (MDH) and NAD-dependent malic enzyme (NAD-ME) activity in Arabidopsis thaliana by integrating reverse genetics, physiological analyses, transcriptomics, quantitative proteomics, and metabolite profiling. Specifically, we generated triple mutants (mdh1xme1xme2) lacking the predominant mitochondrial isoform MDH1 together with both NAD-ME subunits, thereby reducing overall mitochondrial malate conversion capacity. By growing plants under contrasting photoperiod and irradiance regimes to vary photosynthetic demand on malate-linked fluxes, we uncovered a conditional phenotype that was most pronounced under short-day/low-light conditions. Under these conditions, mdh1xme1xme2 exhibited impaired growth and photosynthetic performance, accompanied by cytosolic redox imbalance and altered chloroplast ultrastructure. Transcriptomic profiling revealed that low light unmasks a dawn-phase bottleneck in establishing photosynthetic and redox homeostasis. Consistent with this, the low-light plastid proteome revealed a reallocation away from chloroplast translation and photosynthetic capacity toward proteome maintenance, photoprotection/repair, and iron/ROS management, consistent with a protective acclimation state that nevertheless constrains carbon gain under energy limitation. Low light also triggered C/N imbalance and ammonium accumulation in the mutants. In contrast, increasing irradiance or extending the photoperiod largely alleviated these defects. Together, our results identified mitochondrial malate conversion capacity as a key control point coupling respiratory energy supply and redox homeostasis to photosynthetic metabolism when photosynthetic energy input is limiting.

plant biology↗

bioRxiv2025

Systemic and local regulation of root growth by vascular trehalose 6-phosphate is correlated with re-allocation of primary metabolites between shoots and roots

Trehalose 6-phoshpate (Tre6P) is a key signalling molecule that reflects carbon status and integrates it with developmental decision making. Tre6P has been demonstrated to regulate various developmental processes, including vegetative growth, shoot branching, flowering, and root branching. Here, we investigate how vasculature-derived Tre6P influences root system architecture by expressing heterologous Tre6P synthase (TPS) or Tre6P phosphatase (TPP) specifically in the plant vasculature. Plants with elevated vascular-derived Tre6P levels had smaller root systems, reduced sucrose levels, and lower root metabolite levels, whereas vascular TPP overexpression had the opposite effect. Using reciprocal grafting experiments, we identified shoot-derived vascular Tre6P to be the main driver of these systemic responses. In lines with increased Tre6P in the shoot vasculature, the shoot-to-root metabolite ratios were consistently increased, indicating that Tre6P modulates metabolite allocation and utilization to balance carbon partitioning between shoot and root growth. Our data further suggest that vascular Tre6P contributed to optimizing the carbon-to-nitrogen ratio to support growth and fitness. Besides this systemic function, this study shows that root-derived Tre6P also plays a critical local role in modulating root development. Collectively, our results demonstrate that Tre6P in the vasculature functions both systemically and locally to coordinate metabolic status with root growth and architecture.

plant biology↗

bioRxiv2025

FLOWERING LOCUS T1 is a pleiotropic regulator of reproductive development, plant longevity, and source-sink relations in barley

Source-sink interactions play a critical but mechanistically underexplored role in coordinating reproductive output and longevity in plants. Here, we investigated the role of FT1, the barley homolog of the florigen FLOWERING LOCUS T (FT), in regulating source (leaf) and sink (inflorescence) development and metabolism. Using ft1 knock-out mutants in the spring barley cultivar Golden Promise, which carries a mutated ppd-H1 allele, and in an introgression line with a wild-type Ppd-H1 allele, we show that Ppd-H1 primarily regulates the timing of inflorescence development and flowering through FT1, whereas variation in tiller number and leaf size is determined by the genetic background. ft1 mutants exhibited reduced determinacy of both leaf and inflorescence meristems, resulting in increased leaf and spikelet numbers and size, but severely reduced inflorescence fertility, altered senescence patterns, and significantly extended plant longevity. The ft1 mutants exhibited a strong transcriptional reprogramming of genes involved in both the light and dark reactions of photosynthesis in the leaf, alongside an upregulation of genes associated with carbon catabolism and stress responses in the leaf and inflorescence. Elevated soluble sugar and starch levels in ft1 inflorescences indicated that the impaired development and fertility of ft1 inflorescences were not caused by carbon limitation, but instead reflected a reduced sink strength. Our work reveals that FT1 coordinates the development of vegetative and reproductive meristems and organs with plant physiology and metabolism, thereby regulating source-sink relationships and balancing plant longevity with reproductive output.

plant biology↗

bioRxiv2025

Structural variation at lhcb6 underlies genetic variation in photosystem II maximum quantum efficiency in maize

Targeted utilization of native genetic diversity can expand the genetic basis of traits that exhibit limited genetic variation in elite breeding material and can enhance our understanding of the genotype-phenotype relationships associated with complex traits in crops. In a genome-wide association study in a European maize landrace we identified quantitative trait loci (QTL) that affected the maximum quantum efficiency of photosystem II (Fv/Fm) in field experiments. In a forward genetic approach, we focused on a QTL on chromosome 10, explaining a large proportion of the genetic variance for Fv/Fm in growth stages V4 (35%) and V6 (47%), for genetic dissection and candidate gene discovery. Integrating molecular and physiological information we show that allelic variation at the gene encoding LIGHT HARVESTING CHLOROPHYLL A/B BINDING PROTEIN6 (LHCB6), a component of the photosystem II (PSII) light-harvesting complex (LHCII) antenna, underlies the variation in Fv/Fm. We demonstrate that the allelic variation results from a hAT transposon insertion at lhcb6 and is associated with differential accumulation of the LHCII antenna components LHCB6 and LHCB3, leading to differences in non-photochemical quenching (NPQ) and plant biomass accumulation. Based on proteomic analyses we propose candidate genes that compensate the unfavorable effects caused by impaired LHCII antenna assembly. Our work provides novel insights into the function of lhcb6 in the context of LHCII antenna assembly and demonstrates the value of natural variation in landraces for the understanding and genetic improvement of complex photosynthetic processes.

genetics↗

bioRxiv2025

Impaired lysine biosynthesis drives constitutive energy stress priming and darkness stress responses

Plant survival under prolonged darkness relies on dynamic metabolic reprogramming that redirects limited carbon resources toward mitochondrial respiration and nutrient remobilization. Although amino acids function as alternative respiratory substrates during carbon starvation, how their biosynthesis integrates into this metabolic adjustment remains poorly understood. Here, we investigated the role of lysine biosynthesis using the dapat mutant, which exhibits photoperiod-dependent hypersensitivity to extended darkness. Under short-day conditions, dapat plants exhibited accelerated senescence. reduced photosystem II maximum quantum efficiency, pronounced protein degradation, and accumulation of amino acid, ultimately failing to recover after nine days of darkness. By contrast, survival was largely restored under neutral-day conditions, indicating that restricted carbohydrate reserves during short-day growth exacerbate the mutant phenotype. Remarkably, transcript profiling further revealed that reduced DAPAT activity constitutively activates a catabolic program, including genes associated with amino acid degradation, alternative respiratory pathways, senescence, and autophagy, even under non-stress conditions. Together, these findings identify DAPAT-mediated lysine biosynthesis as a central metabolic hub linking carbon availability to energy and stress signaling. Disruption of this pathway compromises metabolic flexibility and limits the capacity to withstand prolonged carbon starvation. Our results provide new insights into how primary metabolism coordinates stress responses to extended darkness and highlight amino acid biosynthesis as an important component of plant resilience under energy-deprived darkness conditions.

plant biology↗

bioRxiv2025

Multilevel analysis of response to plant growth promoting and pathogenic bacteria in Arabidopsis roots and the role of CYP71A27 in this response

Understanding how plants distinguish between commensal and pathogenic microorganisms is one of the major challenges in the plant microbe interaction research. We previously identified a gene encoding CYP71A27 connected to camalexin, which is necessary for a plant growth promoting (PGP) activity of a number of bacterial strains. To dissect its function, we compared multilevel responses in roots of wild type Arabidopsis and the cyp71A27 mutant to two bacterial strains, a PGP Pseudomonas fluorescens CH267 and a pathogen Burkholderia glumeae PG1. We show that incubation with these bacteria leads to significant and distinct transcriptional reprogramming. This is accompanied by a proteome remodelling in both shoots and roots and profound changes in accumulation of many metabolites, primarily sugars, amino acids, and TCA cycle intermediates, but also by alterations in the ionome. We then analysed the mutant cyp71a27 and identified number of genes and proteins differently regulated, particularly after interaction with the PGP strain, but only a very mild impact of the mutation on root metabolites and exudates. We analysed a variety of mutants in genes differentially regulated by Pseudomonas sp. CH267 in cyp71a27 and revealed that their response to this PGP bacterial strain is similar to cyp71a27. Thus, it seems that CYP71A27 is a non-canonical P-450 without a metabolic function but active in signalling pointing to a regulatory role of the CYP71A27 gene, particularly in interaction with a plant growth promoting bacteria.

plant biology↗

bioRxiv2025

Acylation of the RTX Toxin MbxA stimulates host membrane disruption through a specific interaction with cholesterol

RTX toxins, characterized by their calcium-binding glycine-rich repeat regions, are known for their ability to disrupt host cell membranes. Although the acylation of specific lysine residues in these toxins is established as crucial for their hemolytic activity, the precise mechanisms underlying this enhancement remain unclear. In this study, we explore the role of acylation in the pore-forming behaviour of the RTX toxin MbxA by comparing the lytic activities of acylated MbxA and its non-acylated form, proMbxA, on in vitro membrane systems, as well as living cells. Our findings demonstrate that a cholesterol specific interaction promotes MbxA- induced membrane disruption in an acylation-dependent manner. Further analysis revealed that acylation is not necessary for initial membrane binding but markedly enhances pore formation. Overall, our results offer valuable insights into the molecular determinants that regulate RTX toxin activity, highlighting a complex interplay between lipid composition (sterols), acylation, and membrane disruption, thereby advancing our understanding of RTX toxin pathogenesis.

biochemistry↗

bioRxiv2024

Community-wide interactions sustain life in geothermal spring habitats

We investigated an alga-dominated geothermal spring community in Yellowstone National Park, USA. Our goal was to determine how cells cope with abiotic stressors during diurnal sampling that spanned over two orders of magnitude in solar irradiance. We report a community level response to toxic metal resistance and energy cycling that spans the three domains of life. Arsenic detoxification is accomplished via complementary gene expression by different lineages. Photosynthesis is dominated by Cyanidioschyzon, with the mixotroph, Galdieria, relegated to nighttime heterotrophy. Many key functions, including the cell cycle, are strongly regulated by diurnal light fluctuations. These results demonstrate that biotic interactions are highly structured in extreme habitats. We suggest this was also the case on the early Earth when geothermal springs were cradles of microbial life, prior to the origin of eukaryotes.

evolutionary biology↗