Biology subjects

Mayer, M.

Publications and source records attributed to Mayer, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Characterization of Single Ribosomes and Virus Particles with Large-Diameter Perfringolysin O Nanopores

Existing biological nanopores are often too narrow to accommodate large proteins and biomolecular complexes in their native folded states, precluding the analysis of megadalton assemblies. Here, we report the self-assembly of the cholesterol-dependent cytolysin Perfringolysin O (PFO) into stable transmembrane nanopores composed of 46 {+/-} 9 monomers, with an inner pore diameter of 28.5 {+/-} 5.6 nm and a length of 9.8 nm. We demonstrate that despite the large size of the pore assembly, PFO pores exhibit a stable open-pore current with a high signal-to-noise ratio, making them suitable for resistive-pulse recordings. Moreover, PFO nanopores enable accurate, calibration-free, and label-free sizing of individual proteins, multi-protein complexes, and viral particles across a large molecular-weight range spanning 50 kDa to 3.4 MDa. The exceptionally large diameter of these pores enables, for the first time, resistive-pulse-based characterization of intact virus particles and ribosomes with a biological nanopore. Specifically, we determined the volume, shape, and diameter of complete capsids of recombinant adeno-associated virus serotype 2 (rAAV2) as well as capsid fragments. Finally, simultaneous analysis of molecular volume and shape resolved intact 70S ribosomes from their dissociated 30S and 50S subunits in a mixture. By extending biological nanopore sensing to single-particle characterization of large protein complexes well beyond the reach of existing pores, this approach introduces PFO nanopores as a versatile platform for label-free, single-particle analysis in solution.

biophysics↗

bioRxiv2026

Cryo-EM Structures of Infectious Low-Twist α-Synuclein Filaments in Lewy Bodies

-Synuclein (aSyn) aggregation within Lewy bodies underlies Parkinson's disease and associated dementias. These aggregates are often comprised of amyloid filaments, most of which appear untwisted or have extremely low twist, complicating high-resolution structural determination by cryogenic electron microscopy (cryo-EM). Here, we isolated filaments from the brain tissue of a patient with dementia with Lewy bodies and show infectivity in a cell line system that propagates Lewy aSyn. Then, we determined 2.3-2.9 [A] cryo-EM structures of low-twist aSyn filaments from this tissue and from a publicly available Parkinson's disease dementia dataset. In both structures, the protofilament adopts a fold highly similar to the canonical Lewy fold with conserved cofactor and peptidic densities. Compared to previously resolved high-twist filaments, the low-twist filament is left-handed and flattened along the filament axis, resulting in register shifts of interlayer contacts, while preserving the overall architecture. Our results establish that low-twist filaments, which include over 75% of disease-associated Lewy aSyn amyloids, share a common structural core with high-twist forms and are the predominant aSyn species in samples that engage in templated infection.

biophysics↗

bioRxiv2026

Adaptation of α-synuclein fibrils following multiple system atrophy transmission to mice

Synucleinopathies are a group of neurodegenerative diseases characterized by the presence of misfolded -synuclein inclusions which cause progressive disease by spreading throughout the brain in a prion-like manner. Throughout the neurodegenerative disease field, the ability of a single protein to give rise to multiple distinct clinical disorders is explained by the strain hypothesis, or the idea that the misfolded protein conformation determines the resulting disease. This was initially shown using transmission studies in cell lines and mouse models; more recently cryo-electron microscopy (cryo-EM) validated this idea by identifying distinct -synuclein filament folds in brain tissues from patients with Parkinsons disease, multiple system atrophy (MSA), and juvenile-onset synucleinopathy. However, very little is known about the -synuclein filament structures that form in animal models of these disorders, and thus their relevance to human disease and suitability as models for therapeutic development remains a question. Here we report the first atomic resolution cryo-EM structures of -synuclein fibrils from an MSA patient sample before and after transmission to a transgenic mouse model of disease. Our findings indicate that while distinct adaptations occur during fibril replication in the mouse host, key structural facets are maintained, validating the merits of this transmission model for supporting preclinical research on MSA.

neuroscience↗

bioRxiv2026

Genetic Dissection of Grain Yield and Correlated Proxy Traits Under Suboptimal Conditions

Securing sustainable crop production requires the genetic improvement of abiotic stress tolerance. Due to the broad range of environmental factors causing abiotic stress and complex genotype-by-environment interactions, it is crucial to understand the genetic basis of crop yield under suboptimal conditions. Here, we developed a dent maize Multi-parent Advanced Generation Inter-Cross (MAGIC) population comprising 388 doubled haploid (DH) lines. The population was derived from eight founders with varying stress tolerance, selected from a dent diversity panel evaluated for yield performance across a wide range of European environments. The MAGIC DH lines were genotyped via whole-genome sequencing ([~]5X coverage) and evaluated in seven testcross and 14 line per se trials, for grain dry matter yield, leaf senescence, leaf rolling, anthesis-silking interval, and six additional agronomic traits. Genetic dissection identified 22 grain yield QTL, explaining 45% of the genetic variance. Under heat and drought stress, testcross grain yield correlated significantly with leaf senescence and leaf rolling measured in line per se trials. Bivariate multi-trait analysis showed that alleles for delayed senescence and reduced rolling at detected QTL generally exhibited positive effects on grain yield, suggesting that accumulating these favorable alleles could enhance yield performance. Incorporating these proxies into multi-trait genomic prediction models improved yield prediction accuracy, although gains were constrained by modest trait correlations. Given the comprehensive data, we also provide recommendations for optimizing sequencing depth and QTL mapping strategies in experimental maize populations. Key messageThis eight-founder MAGIC population represents a powerful resource for dissecting complex traits in maize, assessing the utility of drought proxy traits, and optimizing low-coverage whole-genome sequencing approaches.

genetics↗

bioRxiv2026

Circulating Immune Cells are Associated with Non-Inflammatory Pain in Rheumatoid Arthritis

Over half of patients with rheumatoid arthritis (RA) report clinically meaningful pain, despite treatment with disease-modifying antirheumatic drugs (DMARDs). While joint inflammation is a known cause of pain in patients with rheumatic diseases, emerging data indicate that many patients also suffer from centralized or nociplastic pain. There is a critical unmet need to characterize the altered cellular state that distinguishes patients with centralized pain. In the IMPACT study, 39 RA patients with minimal joint inflammation but varying levels of pain underwent quantitative sensory testing (QST) to assess nociplastic pain, completed patient-reported outcome (PRO) surveys, and provided blood samples for immune profiling. Supervised and unsupervised analysis of the multi-parameter spectral flow cytometry data identified immune cell populations correlated with nociplastic pain and patient-reported pain intensity. Moreover, analyses of single-cell RNA-seq from a subset of 22 patients revealed differences in cell type proportions and differential expression between the high and low pain groups. These studies provide novel insights into the role of circulating immune cells in altered central nervous system (CNS) pain regulation in adults with RA.

immunology↗

bioRxiv2025

Classifying Calcium Imaging Dynamics with Deep Learning: Multi-Frequency Analysis through Quantile-Based Time-Series Network Representations

To address the limitations of calcium imaging data, we propose a segmentation-agnostic deep learning framework that integrates Quantile-Based Time-Series Network (QTN) representations with convolutional neural networks to classify neuronal dynamics across multiple spatial resolutions and acquisition frequencies. By transforming fluorescence traces into compact, fixed-size matrices derived from quantile transitions, the method standardizes inputs across recordings while markedly reducing dimensionality and computational cost. Several QTN variants were systematically evaluated, demonstrating strong and consistent classification performance across both whole-image and grid-based preprocessing strategies. Notably, the framework maintained high accuracy under reduced temporal resolution and controlled noise perturbations, confirming that discrimination arises from meaningful temporal patterns rather than artifacts. This study establishes a robust, scalable, and generalizable approach for analyzing calcium imaging dynamics, paving the way for efficient, segmentation-independent characterization of neuronal activity in pharmacological and systems neuroscience applications.

cell 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

Selective and Controllable Trapping of Single Proteins in Nanopores using Reversible Covalent Bonds

Analysis of individual proteins using nanopores makes it possible to determine their size and shape in a label-free approach, within minutes, and from L sample volumes. Short residence times of proteins in the nanopore, high electrical current noise, and bandwidth limitations of the recording electronics during resistive pulse recordings, however, limit the accuracy of size and shape analysis of individual proteins. The work presented here introduces a polymer surface coating of solid-state nanopores to minimize non-specific interactions of proteins with the nanopore wall while functionalizing it covalently with phenylboronic acid (PBA) groups. These PBA groups make it possible to trap selectively glycated proteins by taking advantage of the formation of reversible covalent bonds between PBA and vicinal diol groups of glycated amino acid residues on proteins. Dwell time analysis revealed two populations of resistive pulses: short pulses with dwell times td below 0.4 ms from free translocation of proteins and resistive pulses that we term "long events" that last from 0.4 ms to 2 s and result from intended transient covalent bonds between glycated proteins and PBA groups in the nanopore lumen. The choice of applied potential differences during nanopore recordings or the pH value of the recording buffer makes it possible to control and extend the most probable trapping time of proteins in the nanopore within one to two orders of magnitude. This approach provides the highest accuracy for the determination of protein volume and shape achieved to date with solid-state nanopores and reveals that a trapping time of 1 to 20 ms is ideal to achieve reliable volume and shape analysis while retaining high throughput of the analysis. This approach, hence, extends the residence time of natively glycated proteins or of proteins that are intentionally glycated by straightforward incubation in a glucose solution, thereby providing selectivity and improving the accuracy of nanopore-based characterization of single proteins.

biophysics↗

bioRxiv2025

SPT5 regulates Pol II pausing and elongation in different ways at early versus late embryonic stages

Transcription involves a cycle of initiation, pausing, elongation, and termination. SPT5 regulates both promoter proximal pausing and elongation, but how it orchestrates both steps during dynamic changes in gene expression remains unclear. Here, using Drosophila embryogenesis, we show that pausing both precedes and follows gene expression, while active transcription is accompanied by pause release. Optogenetic rapid depletion of SPT5 from the nucleus uncovered different sensitivities at different developmental stages. In early embryogenesis, SPT5 depletion caused a downstream shift in Pol II pausing to the +1 nucleosome, resulting in defective elongation and early termination. In late embryogenesis, it led to both up- and downregulation of expression, depending on the genes transcriptional and pausing state - upregulation is caused by pause release while downregulation is due to defective elongation. These changes are intensified when genes are increasing or decreasing their transcriptional state, indicating that SPT5 contributes to fine-tuning dynamic changes in gene expression.

genomics↗

bioRxiv2025

Continuous, Low Latency Estimation of the Size and Shape of Single Proteins from Real-Time Nanopore Data

Existing approaches for nanopore sensing typically analyze resistive pulses from the translocation of individual proteins through the nanopore after completing the experiment. This approach foregoes instantaneous protein identification and precludes real-time experimental control. Here, we introduce a method for the analysis of real-time nanopore data capable of characterizing the size and approximate shape of proteins within a millisecond response time during data acquisition. The implemented real-time Two-Sliding Window (TSW) peak detection algorithm makes it possible, for the first time, to filter data, process baselines, and extract resistive pulse information during nanopore recordings using a stream of single data points. This approach achieves a computational throughput of 40 MB/s on a 1.8 GHz laptop CPU. We compared the accuracy of dwell time determination of the TSW algorithm with an established offline threshold searching (TS) algorithm, using simulated resistive pulses. The TSW algorithm accurately extracted events with dwell times greater than 1.5 times the reciprocal of the systems cutoff frequency, 1.5 x (fc)-1. Moreover, the TSW algorithm effectively determined the dwell time of resistive pulses with large intra-event modulation, which is crucial for resolving non-spherical protein-induced fluctuations. Finally, we verify experimentally that integrating the TSW algorithm into the data acquisition process makes it possible to determine the approximate shape and volume of proteins within low-millisecond response times. This real-time data analysis approach potentially provides instant feedback control for manipulating single proteins during or immediately after translocation. This analysis also improves data storage efficiency during recordings with a high sampling rate.

biophysics↗

bioRxiv2025

Cellular composition of tissue-resident monocyte-lineage cells reveal functional heterogeneity during inflammatory arthritis

Tissue-resident monocyte-lineage cell (TRMC) are an extravascular population distinct from circulating monocytes and synovial macrophages and are critical for the development of inflammatory arthritis. However, the precise identities and origins of TRMC subpopulations remain unclear. Here, we characterize the ontogeny of TRMC, which are comprised of bone-marrow (BM)-derived and an embryonic, long-lived population. Furthermore, we identified three TRMC subpopulations, distinguished by expression of TIM4, CX3CR1, and MHCII. Clodronate-laden liposome reduces the number of TRMC but does not impact the proportions or transcriptional profile of TRMC subpopulations at 7 days post administration. TIM4+CX3CR1+ and TIM4+ TRMC represent long-lived population, whereas MHCII+ TRMC are BM-derived and dependent on Ccr2 during steady state. BM-derived TRMC expand and replenish the TIM4+CX3CR1+ and TIM4+ TRMC compartments throughout the peak and plateau of inflammatory arthritis. These findings underscore the importance heterogeneity within TRMC and highlight their distinct responses to synovial disruption and potential roles in rheumatoid arthritis (RA).

immunology↗

bioRxiv2025

Comprehensive analysis of myeloid reporter mice

Macrophages are a pivotal cell type within the synovial lining and sub-lining of the joint, playing a crucial role in maintaining homeostasis of synovium. Although fate-mapping techniques have been employed to differentiate synovial macrophages from other synovial myeloid cells, no comprehensive study has yet been conducted within the mouse synovial macrophage compartment. In this study, we present, for the first time, lineage tracing results from 18 myeloid-specific fate-mapping models in mouse peripheral blood (PB) and synovial tissue. The identification of synovial macrophages and monocyte-lineage cells through flow cytometry was further validated using cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) datasets. These findings provide a valuable methodological tool for researchers to select appropriate models for studying the function of synovial myeloid cells and serve as a reference for investigations in other tissue types.

immunology↗

bioRxiv2025

Pneumolysin nanopores with 20 nm inner diameter to characterize the size and shape of Tau oligomers

Protein nanopores are emerging as versatile tools to fingerprint biomolecules due to their capability to characterize single molecules without the requirement for labeling. A long-standing challenge with biological nanopores is, however, that large biomolecules in their native state are often too large to enter these pores. Here, we report the self-assembly of approximately 35 {+/-} 5 pneumolysin (PLY) toxins to a stable transmembrane pore with a diameter of 20 {+/-} 3 nm, an effective length of 9.5 nm, and excellent low noise characteristics in the context of nanopore-based resistive pulse recordings. The exceptionally large pore diameter enables the characterization of the size and shape of individual proteins and protein complexes ranging in molecular weight from 50 kDa to 0.8 MDa. Moreover, PLY pores make it possible to follow the time course of the formation of oligomers of tau protein in solution by revealing the size, monomer number, approximate shape, and abundance of these oligomers. At least four characteristics make PLY pores well suited for the characterization of heterogeneous amyloid oligomer samples: First, they are not prone to clogging. Second, they provide label-free single particle analysis. Third, their large diameters make it possible to characterize a wide range of amyloid oligomer sizes with high resolution. And fourth, resistive pulse recordings from these pores provide stable open pore current baselines with low electrical noise.

biophysics↗

bioRxiv2025

Human cerebral organoids model tumor infiltration and migration supported by astrocytes in an autologous setting

Efforts to achieve precise and efficient tumor targeting of highly malignant brain tumors are constrained by the dearth of appropriate models to study the effects and potential side effects of radiation, chemotherapy, and immunotherapy on the most complex human organ, the brain. We established a cerebral organoid model of brain tumorigenesis in an autologous setting by overexpressing c-MYC as one of the most common oncogenes in brain tumors. GFP+/c-MYChigh cells were isolated from tumor organoids and used in two different culture approaches: assembloids comprising of a normal cerebral organoid with a GFP+/c-MYChigh tumor sphere and co-culture of cerebral organoid slices at air-liquid interface with GFP+/c-MYChigh cells. GFP+/c-MYChigh cells used in both approaches exhibited tumor-like properties, including overexpression of the c-MYC oncogene, high proliferative and invasive potential, and an immature phenotype as evidenced by increased expression of Ki-67, VIM, and CD133. Organoids and organoid slices served as suitable scaffolds for infiltrating tumor-like cells. Using our highly reproducible and powerful model system that allows long-term culture, we demonstrated that the migratory and infiltrative potential of tumor-like cells is shaped by the environment in which glia cells provide support to tumor-like cells.

cell biology↗

bioRxiv2024

Embracing native diversity to enhance maximum quantum efficiency of photosystem II in maize (Zea mays L.)

Sustainability of maize cultivation would benefit tremendously from early sowing but is hampered by low temperatures during early development in temperate climate. We show that allelic variation of subunit M of NADH-dehydrogenase-like (NDH) complex (ndhm1), discovered in a European maize landrace affects several quantitative traits relevant during early development in cold climates through NDH-mediated cyclic electron transport (CET) around photosystem I, a process crucial for photosynthesis. Starting from a genome-wide association study (GWAS) for maximum potential quantum yield of photosystem II in dark-adapted leaves (Fv/Fm) we capitalized on large phenotypic effects of a hAT transposon insertion in ndhm1 on quantitative traits early plant height (EPH), Fv/Fm, chlorophyll content and cold tolerance caused by reduced protein levels of NDHM and associated NDH components. Analysis of the native allelic series of ndhm1 revealed a rare allele of ndhm1 which is associated with small albeit significant effects on maximum potential quantum yield of photosystem II in dark- and light adapted leaves (Fv/Fm, {Phi}PSII) and early plant height compared to common alleles. Our work showcases the extraction of novel, favorable alleles from locally adapted landraces, offering an efficient strategy for broadening the genetic variation of elite germplasm by breeding or genome editing.

plant biology↗

bioRxiv2024

Differential cell type-specific function of the aryl hydrocarbon receptor and its repressor in diet-induced obesity and fibrosis

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor regulating xenobiotic responses as well as physiological metabolism. Dietary AhR ligands activate the AhR signaling axis in the intestine and throughout the organism, whereas AhR activation is negatively regulated by the AhR repressor (AhRR). While AhR-deficient mice are known to be resistant to diet-induced obesity (DIO), we here demonstrate that AhRR deficiency also leads to a robust, but not as profound protection from DIO and hepatosteatosis. Under conditions of DIO, AhRR-/- mice did not accumulate TCA cycle intermediates in the circulation in contrast to wild-type (WT) mice, indicating protection from metabolic dysfunction. This effect could be mimicked by dietary supplementation of AhR ligands in WT mice. Because of the predominant expression of the AhRR in myeloid cells, AhRR-deficient macrophages were analyzed for changes in metabolism and showed major metabolic alterations regarding oxidative phosphorylation and mitochondrial activity as well as increased expression of genes involved in de novo lipogenesis and mitochondrial biogenesis. Mice with a genetic deficiency of the AhRR in myeloid cells did not show alterations in weight gain after high fat diet (HFD) but demonstrated ameliorated liver damage compared to control mice. Further, deficiency of the AhR in myeloid cells also did not affect weight gain but led to enhanced liver damage and adipose tissue fibrosis compared to controls. Although conditional ablation of either the AhR or AhRR in myeloid cells did not recapitulate the phenotype of the global knockout, our findings suggest that enhanced AhR signaling in myeloid cells deficient for AhRR protects from diet-induced liver damage and fibrosis, whereas myeloid cell-specific AhR deficiency is detrimental.

immunology↗

bioRxiv2023

Comparing sparse inertial sensor setups for sagittal-plane walking and running reconstructions

Estimating spatiotemporal, kinematic, and kinetic movement variables with little obtrusion to the user is critical for clinical and sports applications. Previously, we developed an approach to estimate these variables from measurements with seven lower-body inertial sensors, i.e., the full setup, using optimal control simulations. Here, we investigated if this approach is similarly accurate when using sparse sensor setups with less inertial sensors. To estimate the movement variables, we solved optimal control problems on sagittal plane lower-body musculoskeletal models, in which an objective was optimized that combined tracking of accelerometer and gyroscope data with minimizing muscular effort. We created simulations for 10 participants at three walking and three running speeds, using seven sensor setups with between two and seven sensors located at the feet, shank, thighs, and/or pelvis. We calculated the correlation and root mean square deviations (RMSDs) between the estimated movement variables and those from inverse analysis using optical motion capture (OMC) and force plate data. We found that correlations between IMU- and OMC-based variables were high for all sensor setups, while including all sensors did not necessarily lead to the smallest RMSDs. Setups without a pelvis sensor led to too much forward trunk lean and inaccurate spatiotemporal variables. RMSDs were highest for the setup with two foot-worn IMUs. The smallest setup that estimated joint angles as accurately as the full setup (<1 degree difference in RMSD) was the setup with IMUs at the feet and thighs. The mean correlations for joint angles, moments, and ground reaction forces were at least 0.8 for walking and 0.9 for running when either a pelvic sensor or thigh sensors were included. Therefore, we conclude that we can accurately perform a comprehensive sagittal-plane motion analysis with sparse sensor setups when sensors are placed on the feet and on either the pelvis or the thighs.

bioengineering↗

bioRxiv2023

Structural basis of Gabija anti-phage defense and viral immune evasion

Bacteria encode hundreds of diverse defense systems that protect from viral infection and inhibit phage propagation1-5. Gabija is one of the most prevalent anti-phage defense systems, occurring in >15% of all sequenced bacterial and archaeal genomes1,6,7, but the molecular basis of how Gabija defends cells from viral infection remains poorly understood. Here we use X-ray crystallography and cryo-EM to define how Gabija proteins assemble into an [~]500 kDa supramolecular complex that degrades phage DNA. Gabija protein A (GajA) is a DNA endonuclease that tetramerizes to form the core of the anti-phage defense complex. Two sets of Gabija protein B (GajB) dimers dock at opposite sides of the complex and create a 4:4 GajAB assembly that is essential for phage resistance in vivo. We show that a phage-encoded protein Gabija anti-defense 1 (Gad1) directly binds the Gabija GajAB complex and inactivates defense. A cryo-EM structure of the virally inhibited state reveals that Gad1 forms an octameric web that encases the GajAB complex and inhibits DNA recognition and cleavage. Our results reveal the structural basis of assembly of the Gabija anti-phage defense complex and define a unique mechanism of viral immune evasion.

microbiology↗