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Bayer, E.

Publications and source records attributed to Bayer, E..

10 recordsLinked to original sources

Manipulation of REMORIN gene dosage affects salt signaling and tolerance in Arabidopsis thaliana

Salinity is among the most detrimental environmental plant stressors hampering growth, development and immunity. Its impact is increasing due to intensive agricultural practices and climate change. The molecular components linked to the early cellular and molecular events triggered by salt are mostly unknown. Here we show that genes encoding for members of the REMORIN proteins family exhibit dynamic temporal regulation following salt perception. Analogously to pattern-triggered immune signaling, we observed that salt rapidly induces the reorganization of REMORIN1.2 (REM1.2) into static plasma membrane nanodomains which colocalize with the actin-nucleating protein FORMIN 6. Overexpression of REM1.2 inhibits early signaling and late cell morphological responses to salt, defects which correlated with an increase in salt sensitivity. Our findings connect early salt-triggered signaling events to changes in cell architecture and root resilience and describe REMORIN nanodomains as a convergent articulation point in biotic and abiotic signaling pathways.

plant biology↗

Mucinolysome in gut microbiomes of farm animals and humans

Mucins are glycoproteins that create a protective barrier protecting host tissues from microbial pathogens and are instrumental for host health. Here, we provide evidence that mucin glycan degradation in the gut can be mediated by mucinolysomes, defined as extracellular multi-enzyme complexes specializing in mucin glycan degradation. We computationally predicted the presence of mucinolysomes across 63 metagenome-assembled genomes (MAGs) and two isolated genomes of anaerobic Limousia bacteria, including seven MAGs from human samples of six countries. All 65 genomes were found to display core mucinolysome components, consisting of 3[~]6 scaffoldins (containing up to 12 cohesin modules) and up to 22 dockerin-containing mucin glycan-degrading CAZymes (carbohydrate active enzymes). The organization of mucinolysomes allows the assembly of up to 24 CAZymes in the same complex. We validated that a cultivated Limousia strain ET540 from chicken cecum can support growth on mucins as its sole carbon source, triggering the expression of most mucinolysome-related genes, including both scaffoldins and CAZymes. We also modeled the assembly of proteins into a multi-enzyme complex by predicting the cohesin-dockerin interactions among most of the mucinolysome proteins using AlphaFold3. While mucinolysosome-encoding Limousia have low abundance in different animal hosts, their abundance and prevalence are higher in farm animals than in humans, highlighting a potentially important role in livestock gut ecosystems. Our findings reveal a novel mechanism of mucin glycan degradation and provide a framework to explore microbial contributions to gut health and host-microbe interactions across species.

bioinformatics↗

Morphologically Tunable Mycelium Chips for Physical Reservoir Computing

AO_SCPLOWBSTRACTC_SCPLOWWe introduce a neuromorphic computing substrate based on PEDOT:PSS-infused mycelium, a biofabricated, morphologically tunable material that can be engineered into electrically active components including resistors, capacitors, and non-linear elements. Leveraging the principles of physical reservoir computing, we demonstrate that hyphal networks grown under controlled environmental conditions can transform time-varying inputs into nonlinear, high-dimensional state trajectories, enabling machine learning tasks such as NARMA-10 sequence prediction. The chips are produced using a "design-grow-compute" workflow that integrates morphological modeling, parametric growth protocols, and vacuum-assisted polymer infusion. Morphological complexity is shown to influence charge transport and memory capacity, offering a new axis of control for designing analog computational architectures. Our prototype chips interface with a custom carrier board enabling analog signal conditioning and readout. Benchmarking revealed robust nonlinearity, temporal dynamics, and task-relevant separability. Unlike memristor arrays, photonic, or living-cell-based reservoir systems, our non-living analog mycelium chip is low-cost, biodegradable, and scalable using existing mushroom farming infrastructure, with production yields exceeding 3 million chips per growth cycle. This proof-of-concept demonstration using a single prototype device establishes the first biodegradable reservoir computing platform, with performance trade-offs justified by unprecedented sustainability advantages and orders-of-magnitude cost reductions. This advances a novel direction for biologically derived, single-use (compostable) or very large-scale machine learning hardware and introduces mycelium as a functional medium for analog inference.

bioengineering↗

Root Expansion Microscopy (ROOT-ExM): A streamlined super resolution method for plants

Expansion microscopy (ExM) has revolutionized biological imaging by physically enlarging samples, surpassing the light diffraction limit and enabling nanoscale visualization using standard microscopes. While extensively employed across a wide range of biological samples, its application to plant tissues is sparse. In this work, we present ROOT-ExM, an expansion method suited for stiff and intricate multicellular plant tissues, focusing on the primary root of Arabidopsis thaliana. ROOT-ExM achieves isotropic expansion with a fourfold increase in resolution, enabling super-resolution microscopy comparable to STimulated Emission Depletion (STED) microscopy. Labelling is achieved through immunolocalization, compartment-specific dyes, and native fluorescence preservation, while N-Hydroxysuccinimide (NHS) ester-dye conjugates reveal the ultrastructural context of cells alongside specific labelling. We successfully applied ROOT-ExM to image various cellular structures, including the Golgi apparatus, the endoplasmic reticulum, the cytoskeleton, and wall-embedded structures such as plasmodesmata. When combined with lattice light sheet microscopy (LLSM), ROOT-ExM achieves 3D quantitative analysis of nanoscale cellular process, revealing increased vesicular fusion in close proximity of the cell plate during cell division. Achieving super-resolution fluorescence imaging in plant biology remains a formidable challenge. Our findings underscore that ROOT-ExM provides a remarkable, cost-effective solution to this challenge, paving the way for unprecedented insights into plant cellular subcellular architecture. One sentence summaryROOT-ExM achieves super-resolution expansion microscopy in plants

plant biology↗

Plasmodesmata act as unconventional membrane contact sites regulating inter-cellular molecular exchange in plants

Membrane contact sites (MCS) are fundamental for intracellular communication, but their role in intercellular communication remains unexplored. We show that in plants, plasmodesmata communication bridges function as atypical endoplasmic reticulum (ER)-plasma membrane (PM) tubular MCS, operating at cell-cell interfaces. Similar to other MCS, ER-PM apposition is controlled by a protein-lipid tethering complex, but uniquely, this serves intercellular communication. Combining high-resolution microscopy, molecular dynamics, pharmacological and genetic approaches, we show that cell-cell trafficking is modulated through the combined action of Multiple C2 domains and transmembrane domain proteins (MCTP) 3, 4, and 6 ER-PM tethers, and phosphatidylinositol-4-phosphate (PI4P) lipid. Graded PI4P amounts regulate MCTP docking to the PM, their plasmodesmata localization and cell-cell permeability. SAC7, an ER-localized PI4P-phosphatase, regulates MCTP4 accumulation at plasmodesmata and modulates cell-cell trafficking capacity in a cell-type specific manner. Our findings expand MCSs functions in information transmission, from intracellular to intercellular cellular activities. In briefPlant intercellular communication is regulated via tubular membrane contact through PI4P binding-ER-PM tether MCTP proteins HighlightsO_LIPlasmodesmata are unconventional ER/PM tubular contact sites located at cell-cell interface C_LIO_LIPlasmodesmata operate as control valves, modulating ER-PM contacts to regulate transport C_LIO_LIMCTP3, MCTP4, MCTP6 and PI4P tethering elements act as valve regulators C_LIO_LISAC7 PI4P phosphatase controls plasmodesmata MCS in a cell-type-specific manner C_LI

plant biology↗

A hetero-oligomeric remorin-receptor complex regulates plant development

Plant growth and development are modulated by both biotic and abiotic stress. Increasing evidence suggests that cellular integration of the corresponding signals occurs within preformed hubs at the plasma membrane called nanodomains. These membrane sub-compartments are organized by multivalent molecular scaffold proteins, such as remorins. Here, we demonstrate that Group 1 remorins form a hetero-oligomeric complex at the plasma membrane. While these remorins are functionally redundant for some pathways their multivalency also allows the recruitment of specific interaction partners. One of them, the receptor-like kinase REMORIN-INTERACTING RECEPTOR 1 (RIR1), that acts redundantly with the closely related receptor NILR2, is specifically recruited by REM1.2 in a phosphorylation-dependent manner. Overlapping developmental phenotypes suggest that the REM/RIR complex regulates key developmental pathways.

plant biology↗

Surface display of designer protein scaffolds on genome-reduced strains of Pseudomonas putida

The bacterium Pseudomonas putida KT2440 is gaining considerable interest as a microbial platform for biotechnological valorization of polymeric organic materials, such as waste lignocellulose or plastics. However, P. putida on its own cannot make much use of such complex substrates, mainly because it lacks an efficient extracellular depolymerizing apparatus. We seek to meet this challenge by adopting a recombinant cellulosome strategy for this attractive host. Here, we report an essential step in this endeavor – a display of designer enzyme-anchoring protein “scaffoldins”, encompassing cohesin binding domains from divergent cellulolytic bacterial species on the P. putida surface. Two P. putida chassis strains, EM42 and EM371, with streamlined genomes and substantial differences in the composition of the outer membrane were employed in this study. Scaffoldin variants were delivered to their surface with one of four tested autotransporter systems (Ag43 from Escherichia coli), and the efficient display was confirmed by extracellular attachment of chimeric β-glucosidase and fluorescent proteins. Our results highlight the importance of cell surface engineering for display of recombinant proteins in Gram-negative bacteria and pave the way towards designer cellulosome strategies, tailored for P. putida.Competing Interest StatementThe authors have declared no competing interest.View Full Text

synthetic biology↗

Dynamic interactions of type I cohesin modules fine-tune the structure of the cellulosome of Clostridium thermocellum

Efficient degradation of plant cell walls by selected anaerobic bacteria is performed by large extracellular multienzyme complexes termed cellulosomes. The spatial arrangement within the cellulosome is organized by a protein called scaffoldin, which recruits the cellulolytic subunits through interactions between cohesin modules on the scaffoldin and dockerin modules on the enzymes. Although many structural studies of the individual components of cellulosomal scaffoldins have been performed, the role of interactions between individual cohesin modules and the flexible linker regions between them are still not entirely understood. Here, we report single-molecule measurements using Forster resonance energy transfer to study the conformational dynamics of a bimodular tandem cohesin segment of the scaffoldin protein CipA of Clostridium thermocellum. Our data reveal the existence of compacted structures in solution that persist on the timescale of milliseconds. The compacted conformation is found to be in dynamic equilibrium with an extended state that shows distance fluctuations on the microsecond timescale. Shortening of the inter-cohesin linker does not significantly alter the structural dynamics. Upon addition of dockerin-containing enzymes, an extension of the flexible state is observed but the cohesin-cohesin interactions persist. This suggests that the dockerin-binding interfaces are not involved in cohesin-cohesin interactions. The formation of cohesin-cohesin interactions is also observed in all-atom molecular dynamics simulations of the system. From the simulations, we identify possible inter-cohesin binding modes, none of which show obstruction of the cohesin-dockerin binding interfaces. Our results go beyond the view of scaffoldin as \"beads on a string\". We propose that both the flexibility and cohesin-cohesin interactions are important factors for the precise spatial arrangement of the enzymatic subunits in the cellulosome that leads to the high catalytic synergy in these assemblies. Hence, the flexibility of the linker region and cohesin-cohesin interactions should be considered when designing cellulosomes for industrial applications.

biophysics↗

An atlas of Caenorhabditis elegans chemoreceptor expression

One goal of modern day neuroscience is the establishment of molecular maps that assign unique features to individual neuron types. Such maps provide important starting points for neuron classification, for functional analysis and for developmental studies aimed at defining the molecular mechanisms of neuron identity acquisition and neuron identity diversification. In this resource paper, we describe a nervous system-wide map of the potential expression sites of 244 members of the largest gene family in the C. elegans genome, rhodopsin-like (class A) GPCR chemoreceptors, using classic gfp reporter gene technology. We cover representatives of all sequence families of chemoreceptors GPCRs, some of which were previously entirely uncharacterized. Most reporters are expressed in a very restricted number of cells, often just in single cells. We assign GPCR reporter expression to all but two of the 37 sensory neuron classes of the sex-shared, core nervous system. Some sensory neurons express a very small number of receptors, while others, particularly nociceptive neurons, co-express several dozen GPCR reporter genes. GPCR reporters are also expressed in a wide range of inter- and motorneurons, as well as nonneuronal cells, suggesting that GPCRs may constitute receptors not just for environmental signals, but also for internal cues. We observe only one notable, frequent association of coexpression patterns, namely in one nociceptive amphid (ASH) and two nociceptive phasmid sensory neurons (PHA, PHB). We identified GPCRs with sexually dimorphic expression and several GPCR reporters that are expressed in a left/right asymmetric manner. We identified a substantial degree of GPCR expression plasticity; particularly in the context of the environmentally-induced dauer diapause stage when one third of all tested GPCRs alter the cellular specificity of their expression within and outside the nervous system. Intriguingly, in a number of cases, the dauer-specific alterations of GPCR reporter expression in specific neuron classes are maintained during postdauer life and in some case new patterns are induced post-dauer, demonstrating that GPCR gene expression may serve as traits of life history. Taken together, our resource provides an entry point for functional studies and also offers a host of molecular markers for studying molecular patterning and plasticity of the nervous system.\n\nAUTHOR SUMMARYMaps of gene expression patterns in the nervous system provide an important resource for neuron classification, for functional analysis and for developmental studies that ask how different neurons acquire their unique identities. By analyzing transgenic gfp reporter strains, we describe here the expression pattern of 244 putative chemosensory receptor-encoding genes, which constitute the largest gene family in C.elegans. We show that, as expected, chemoreceptor expression is enriched in chemosensory neurons but it is also expressed in a wide range of interneurons, motorneurons, as well as non-neuronal cells, suggesting that putative chemosensory receptors may not just sense environmental signals but also internal cues. We find that each chemoreceptor is expressed in a few neuron types, often just one, but each neuron type can express a large number of chemoreceptors. Interestingly, we uncovered that chemoreceptor expression is remarkably plastic, particularly in the context of the environmentally-induced dauer diapause stage. Taken together, this molecular atlas of chemosensory receptors provides an entry point for functional studies and offers a host of markers for studying neuronal patterning and plasticity.

genetics↗

The plant calcium-dependent protein kinase CPK3 phosphorylates REM1.3 to restrict viral infection

Plants respond to pathogens through dynamic regulation of plasma membrane-bound signaling pathways. To date, how the plant plasma membrane is involved in responses to viruses is mostly unknown. Here, we show that plant cells sense the Potato virus X (PVX) COAT PROTEIN and TRIPLE GENE BLOCK 1 proteins and subsequently trigger the activation of a membrane-bound calcium-dependent kinase. We show that the Arabidopsis thaliana CALCIUM-DEPENDENT PROTEIN KINASE 3-interacts with group 1 REMORINs in vivo, phosphorylates the intrinsically disordered N-terminal domain of the Group 1 REMORIN REM1.3, and restricts PVX cell-to-cell movement. REM1.3-s phospho-status defines its plasma membrane nanodomain organization and is crucial for REM1.3-dependent restriction of PVX cell-to-cell movement by regulation of callose deposition at plasmodesmata. This study unveils plasma membrane nanodomain-associated molecular events underlying the plant immune response to viruses.

plant biology↗