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This feed contains articles for bioRxiv Subject Collection "All"
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<title>bioRxiv</title>
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<link>https://www.biorxiv.org</link>
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<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747358v1?rss=1">
<title>
<![CDATA[
Bayesian adaptive experimental design for efficient microbial genome-wide association studies 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747358v1?rss=1
</link>
<description><![CDATA[
Bacterial genome-wide association studies (GWAS) offer a powerful approach to identify the genetic basis of a trait measured in a set of sequenced isolates. As the number of sequenced isolates has grown, the limiting factor for GWAS has become phenotyping enough isolates to achieve statistical power. To overcome the need for large-scale phenotyping, we developed Bayesian Adaptive Sequential Sampling GWAS (BASS-GWAS), which couples Bayesian adaptive experimental design with a sparse regression model to select maximally informative isolates for phenotypic testing. BASS-GWAS efficiently recovered causal loci for three antimicrobial resistance traits in Neisseria gonorrhoeae, requiring many fewer phenotyped isolates than random sampling. We applied BASS-GWAS to discover variants enabling gyrBD429N-dependent cross-resistance to the novel topoisomerase inhibitors zoliflodacin and gepotidacin. After phenotyping fewer than 30 isolates, we identified and then validated both parCD86N and a gyrA-parE-based pathway as enabling cross-resistance. BASS-GWAS provides a practical and statistically principled solution for efficient bacterial GWAS.
]]></description>
<dc:creator><![CDATA[ Helekal, D., Blomqvist, S. O. P., Mukherjee, A., Bowcutt, B. A., Palace, S. G., Grad, Y. H. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747358</dc:identifier>
<dc:title><![CDATA[Bayesian adaptive experimental design for efficient microbial genome-wide association studies]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747232v1?rss=1">
<title>
<![CDATA[
Simvastatin attenuates disease phenotypes in human induced pluripotent stem cell models of familial Parkinson's disease through RhoA inhibition 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747232v1?rss=1
</link>
<description><![CDATA[
Background: Statins have gained increasing interest for their potential therapeutic effect in Parkinson's disease (PD). Beyond their cholesterol-lowering effect, statins decrease synthesis of isoprenoids, which is believed to account for their pleiotropic effects. Isoprenylation is important for proper membrane localization and function of the Rho GTPases, including RhoA. RhoA signalling has emerged as a possible underlying signalling pathway involved in the pathogenesis of PD and other neurodegenerative diseases. Methods: In the present study, we investigated the effects of simvastatin on neurodegeneration-associated phenotypes using human induced pluripotent stem cell-derived dopaminergic (DA) neurons from both PD patients and isogenic PARK2-/- cell lines. The dependence on RhoA was confirmed using direct RhoA inhibition using rhosin. Assessed phenotypes included structural integrity, mitochondrial and lysosomal characteristics, cytokine secretion, and cell viability. To understand the relevance of RhoA in PD, RhoA activity was measured in 32 PD patient iPSC-derived lines with different familial PD-related mutations and in healthy controls. Results: Simvastatin rescued multiple PD-associated phenotypes, including impaired DA neurite outgrowth, mitochondrial and lysosomal alterations, cytokine release, and cell death. RhoA inhibition was associated with changes in mitophagy- and autophagy-related markers, suggesting improved autophagic and mitophagic turnover. Furthermore, we performed the first systematic screen of RhoA activity across 32 iPSC-derived DA neuron lines representing multiple genetic forms of PD (PINK1 loss of function, parkin loss of function, LRRK2 (G2019S), LRRK2 (R1441C), GBA (L44P), GBA (N370S), A53T, and SNCA triplication) and healthy controls. RhoA activity was perturbated across several genetic forms of PD subtypes and was significantly increased in many, although not all, patient lines compared with healthy controls, highlighting disease heterogeneity and supporting RhoA dysregulation as a shared pathogenic mechanism in a subset of PD. Conclusions: Our findings identify aberrant RhoA signalling as a convergent pathogenic mechanism across multiple forms of genetic PD and demonstrate that simvastatin ameliorates PD-associated phenotypes through RhoA inhibition. These results support RhoA as a promising therapeutic target while emphasizing the importance of patient stratification based on RhoA activity.
]]></description>
<dc:creator><![CDATA[ Schmidt, S. I., Okarmus, J., Ryding, M., Skousen, I. K., Broner Jensen, N. F., Christensen, E. B., Winkelmann, L. S., Juhl, A. D., Klaebel, M., Blaabjerg, M., Freude, K., Wustner, D., Wade-Martins, R., Ryan, B., Meyer, M. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747232</dc:identifier>
<dc:title><![CDATA[Simvastatin attenuates disease phenotypes in human induced pluripotent stem cell models of familial Parkinson's disease through RhoA inhibition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.28.747825v1?rss=1">
<title>
<![CDATA[
Ratiometric growth-rate control enables robust coexistence in competing microbial consortia 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.28.747825v1?rss=1
</link>
<description><![CDATA[
Maintaining a prescribed composition in engineered microbial consortia is difficult because small fitness differences can drive competitive exclusion. We study a two-strain consortium in continuous culture and develop a feedback architecture that regulates composition by selectively slowing the fast strain as a function of the population ratio. At the population level, we derive an idealized ratio-feedback law with a tunable positive coexistence equilibrium. We then propose a biomolecular realization using orthogonal quorum sensing, an sRNA-based ratiometric controller, and a ppGpp-mediated growth actuator. Exploiting the separation between slow population growth and faster intracellular controller dynamics, we use singular perturbation theory to show that, for sufficiently fast controller dynamics, the full implementation model inherits the coexistence equilibrium and its local stability properties from the reduced model. Numerical simulations validate the reduction and show how weaker timescale separation or loss of the assumed molecular regime degrades performance.
]]></description>
<dc:creator><![CDATA[ Barajas, C. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.28.747825</dc:identifier>
<dc:title><![CDATA[Ratiometric growth-rate control enables robust coexistence in competing microbial consortia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.743980v1?rss=1">
<title>
<![CDATA[
Shape Analysis of Coronary Flow Waveforms using Singular Value Decomposition 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.743980v1?rss=1
</link>
<description><![CDATA[
Coronary flow waveforms have a distinct diastolic-dominant shape with periods of low or retrograde flow during systole. While the general waveform shape has been attributed to complex interactions between cardiac and vascular mechanics, there is limited research into the variability in coronary flow waveforms and what this variability may reveal about cardiac function. This work presents a shape analysis of left anterior descending artery (LAD) flow waveforms using Fourier transforms and Singular Value Decomposition (SVD) performed on baseline data collected from 32 pigs. Pigs included in the study reflect two breeds (Ossabaw and Yorkshire) and three different experimental conditions (lean-control, lean-paced, and obese-paced). Fourier transforms were used to decompose the waveforms into 15 harmonics for each pig. An SVD analysis is then used to extract temporal patterns of the waveforms. Correlations between pig-specific coefficients for the SVD modes and clinical metrics were used to investigate physiological explanations of LAD waveform variability. Temporal LAD flow patterns of the second SVD mode are significantly correlated with heart rate. The third SVD mode significantly correlates with mean blood pressure and maximum hyperemic flow. Furthermore, the fourth SVD mode is weakly correlated with left-ventricular end diastolic pressure and endocardial-epicardial flow ratios. This work demonstrates that LAD flow waveforms can be broken down into temporal patterns that correlate with physiological features. Furthermore, this shape-analysis method allows for waveform reconstruction and simplifies visualization of the temporal patterns identified using SVD, an advantage over existing methods that focus on characterizing flow waveforms by points of interest.
]]></description>
<dc:creator><![CDATA[ Sturgess, V. E., Schenk, N. A., Ziegele, J. W., Essajee, S. I., Tune, J. D., Rajapakse, I., Figueroa, C. A., Beard, D. A. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.743980</dc:identifier>
<dc:title><![CDATA[Shape Analysis of Coronary Flow Waveforms using Singular Value Decomposition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748161v1?rss=1">
<title>
<![CDATA[
Structure of an RNA polymerase ribozyme replication complex 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748161v1?rss=1
</link>
<description><![CDATA[
Life began with the emergence of a molecule that could replicate its own genetic material, a task plausibly mediated by an RNA-dependent RNA polymerase ribozyme. Here, we present the structure of such a polymerase ribozyme, bound to RNA substrates comprising the template, primer, and nucleoside triphosphate (NTP) analog. The structure reveals how directed evolution shaped flanking elements around a highly conserved catalytic core derived from the ancestral class I ligase ribozyme. Each element serves as a functional module, positioning the primer-template duplex and incoming NTP within the active site of the enzyme. This emergent domain organization is remarkably similar to the "right hand" configuration of polymerase proteins, suggesting a common functional form for copying nucleic acids, regardless of biopolymer catalyst.
]]></description>
<dc:creator><![CDATA[ Strutzenberg, T. S., Horning, D. P., Cochrane, W. G., Andrade, L., Han, X., Joyce, G. F., Lyumkis, D. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748161</dc:identifier>
<dc:title><![CDATA[Structure of an RNA polymerase ribozyme replication complex]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747338v1?rss=1">
<title>
<![CDATA[
Network-based meta-analysis maps stage-dependent molecular programs in MASLD through MASLD-META NETWORK application 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747338v1?rss=1
</link>
<description><![CDATA[
Metabolic dysfunction-associated steatotic liver disease (MASLD), the leading cause of chronic liver pathologies worldwide, represents a growing clinical burden. Its diagnosis remains reliant on liver biopsy that limits early detection and the ability to capture molecular changes across disease progression. A systematic understanding of stage-dependent gene expression changes is essential to identify biomarkers and effectively characterize disease mechanisms. Therefore recent studies provided databases for searching genes as well as prediction of multi-gene signatures for disease progression. However, there is still a need for interactive and comprehensive meta-analysis of datasets of MASLD patients with available histological metadata. Herein, we performed a meta-analysis of RNA-seq datasets using NAFLD Activity Score (NAS; n = 897) and fibrosis stage (n = 856) upon conducting pairwise comparisons across histological stages and identified differentially expressed genes associated with disease progression. Most importantly, we provide our findings via a dedicated web server, the MASLD-META NETWORK (https://masld.scilicium.com), enabling users to interactively explore meta-analysis results across diverse network modalities. In addition, we characterized gene expression dynamics across increasing disease stages to identify consistent progression-associated pathways using Louvain clustering. Network-based parameters such as centrality in combination with meta-analysis scores further highlighted central genes and pathways implicated in disease mechanisms. Accordingly, MASLD-META NETWORK enabled an integrative reassessment of recently published gene signatures, identifying COL1A1, COL3A1, THBS2, FBLN5, and PDGFA as the most central genes, and SULF2, MMP14, IL32, GPNMB, and COL3A1 as candidate markers of earlier transcriptional alterations. Network analysis of MASLD associated biological modules further identified LAMA2 and LAMA3 as previously unrecognized central candidate targets.
]]></description>
<dc:creator><![CDATA[ Kumak, E., Darde, T., Konu, O. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747338</dc:identifier>
<dc:title><![CDATA[Network-based meta-analysis maps stage-dependent molecular programs in MASLD through MASLD-META NETWORK application]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747388v1?rss=1">
<title>
<![CDATA[
MetaDome 2027: a comprehensively updated resource for aggregating missense variant evidence across homologous human protein domains 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747388v1?rss=1
</link>
<description><![CDATA[
The interpretation of missense variants remains a major challenge in clinical genetics. "Meta-domains" aggregate population and pathogenic variation across homologous Pfam domain instances in the human proteome, providing per-residue context for interpreting variants of uncertain significance (VUS). Our 2019 implementation, MetaDome, is widely used and named in clinical variant-classification guidelines. Here we present the MetaDome 2027 update, featuring a comprehensively updated dataset and GRCh38 support. The redesigned pipeline enables incremental updates of GENCODE, UniProtKB/Swiss-Prot, Pfam, gnomAD, and ClinVar while maintaining 100% sequence-identity gene-to-protein mapping. Annotated Pfam domain instances grew 14.9% from 71,419 to 82,069 and meta-domain-eligible Pfam families ([&ge;]2 human occurrences) by 73.3% from 3,334 to 5,778; Pfam domains are annotated to 92% of human proteins. Approximately 43% of mapped protein-coding nucleotides (14.3 million in GRCh38, 13.8 million in GRCh37) are in a meta-domain; in GRCh38 67.9% (37,692 of 55,548) of pathogenic or likely pathogenic ClinVar missense variants fall at such a position. We show how MetaDome helped reclassify a de novo missense VUS in RALA and identify 52,463 ClinVar missense VUS for which meta-domains supply otherwise unavailable pathogenic evidence. MetaDome is freely available at www.metadome.app.
]]></description>
<dc:creator><![CDATA[ Wiel, L., Ferraro, F., Yu, J., Zhen, J., Nachun, D., Mendez, R., Reuter, C. M., Cui, J. L., Bonner, D. E., Carter, J. N., Marwaha, S., van de Vorst, M., Emami, S., Kravets, E., Neu, M. B., van Ham, T. W., Kleefstra, T., Ashley, E. A., Bernstein, J. A., Montgomery, S. B., Gilissen, C., Wheeler, M. T. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747388</dc:identifier>
<dc:title><![CDATA[MetaDome 2027: a comprehensively updated resource for aggregating missense variant evidence across homologous human protein domains]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747253v1?rss=1">
<title>
<![CDATA[
Every Cure Knowledge Graph: A Unified Biomedical Knowledge Graph for Drug Repurposing 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747253v1?rss=1
</link>
<description><![CDATA[
Identifying causal connections between existing drugs and mechanistic profiles of diseases is a foundational step for effective drug repurposing. Although knowledge graphs (KGs) are highly suited for consolidating biomedical databases and tracking these connections, a single biomedical KG is constrained by its ingestion pipeline and knowledge sources. While different biomedical KGs could be complementary if combined, efforts to combine them into a unified and more comprehensive KG are hindered by lack of interoperability and poor provenance. To address those issues, we present EC-KG, a Biolink Model-compatible KG for computational drug repurposing. EC-KG is an interoperable, provenance-first KG which integrates RTX-KG2, ROBOKOP, and PrimeKG at the network-level, encapsulating over 7 million nodes and 81 million edges from 95 primary data sources. EC-KG has improved coverage of core biomedical entities such as drugs, targets, and diseases relevant to drug repurposing vs source graphs, and captures complex biomedical mechanisms within its topology. We demonstrate that the network unification in EC-KG leads to emergence of novel, mechanistically relevant pathways which are disconnected in the underlying constituent networks and show its applications in method development, benchmarking and predictive drug repurposing applications. EC-KG has already been successfully used in drug repurposing research to surface Botulinum Toxin A as a candidate to treat Major Depressive Disorder, as well as to validate repurposing of Lenalidomide and Dexamethasone for a subgroup of patients with Rosai-Dorfman Disease.
]]></description>
<dc:creator><![CDATA[ Kaniewski, P., Carter, E. K., Rhodes, D., Lim, E. M., Li, J., Vergine, J., Matentzoglu, N., Schaper, K., Reilly, J., Sundar, S., Vijnck, L., Sharp, E., Alfonso, N., Ford, A., Stepanenko, A., Hempstead, C., Brokmeier, P., Bizon, C., Tropsha, A., Haendel, M. A., Fajgenbaum, D. C., Lancashire, L. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747253</dc:identifier>
<dc:title><![CDATA[Every Cure Knowledge Graph: A Unified Biomedical Knowledge Graph for Drug Repurposing]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.31.747927v1?rss=1">
<title>
<![CDATA[
The evolutionarily conserved C-terminal domain of a domesticated transposase-derived protein regulates its DNA integration ability 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.31.747927v1?rss=1
</link>
<description><![CDATA[
THAP9 is a transposable element-derived gene which encodes a protein that is homologous to the active Drosophila P-element transposase (DmTNP). Both THAP9 and DmTNP possess a C-terminal domain (CTD) which is functionally uncharacterized. Sequence and structural analysis suggest that the THAP9-CTD has a novel fold which is only found in THAP9 homologs. To explore the evolutionary history and characteristics of this novel domain, exhaustive phylogenetic analysis (using MSA, structure prediction, MSTA-based clustering) was performed. THAP9-CTD homologs were more widely distributed throughout the animal kingdom in comparison to DmTNP-CTD homologs which were restricted to arthropods. Moreover, the THAP9-CTD homologs were more conserved, especially among mammals and birds and their average length increased in a class-specific manner. Comparison with the DmTNP-CTD homologs demonstrates that although their respective CTDs may have evolved independently, they both surprisingly share similar secondary structure elements consisting of three conserved helical regions made of hydrophobic residues that are predicted to make up a conserved core. The role of the respective CTDs were further investigated by creating truncation mutants lacking the CTD. Interestingly both THAP9 and DmTNP truncation mutants are still capable of DNA excision and integration suggesting that their respective CTDs are not essential for DNA transposition. Moreover, CTD truncation favours DNA integration in THAP9: this suggests that CTD acquisition during evolution may have led to THAP9 domestication as observed in other transposable element-derived genes like Rag1 and piggybac, which have similar terminal regulatory domains.
]]></description>
<dc:creator><![CDATA[ Saha, A., Ghosh, A., Majumdar, S. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.31.747927</dc:identifier>
<dc:title><![CDATA[The evolutionarily conserved C-terminal domain of a domesticated transposase-derived protein regulates its DNA integration ability]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747241v1?rss=1">
<title>
<![CDATA[
Higher rewards lead to more accurate flower detection and increased contrast sensitivity in the bumblebee Bombus terrestris 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747241v1?rss=1
</link>
<description><![CDATA[
In vertebrates, top-down visual attention is a cognitive process where internal goals modulate the tuning of peripheral sensory systems. This leads to increased perceived contrast to both goal-relevant objects and areas of the visual field that are attended. Such a system would also be beneficial to bees, enabling them to detect and recognise the most profitable flowers in their environment. We tested whether bumblebees possess a top-down attentional system resembling that seen in vertebrates. We trained two groups of bees to collect rewards under high contrast targets. To potentially induce a difference in attention while searching for the targets, one group received a higher concentration of sucrose rewards compared to the other. During tests, the targets were presented with a series of lower contrasts to measure the contrast sensitivity curves of the bees induced by the different learnt reward levels. We predicted a stronger effect of any attention-like process on contrast sensitivity in the high reward group. We also repeated this experiment with the neonicotinoid pesticide imidacloprid dissolved in the sucrose rewards to test whether this affects bee attention. Across all test contrasts, higher rewards significantly increased bee accuracy when locating targets, lowered contrast thresholds and reduced the latency to make first choices. Imidacloprid reduced bee accuracy but did not influence first choice latency. These results suggest that learnt floral rewards can influence bee behavioural contrast sensitivity in a manner resembling vertebrate top-down attention and that imidacloprid may modulate this through effects on their nervous system.
]]></description>
<dc:creator><![CDATA[ Robert, T., Flett, E., Le Lay, H., Nicolas, M., Nityananda, V. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747241</dc:identifier>
<dc:title><![CDATA[Higher rewards lead to more accurate flower detection and increased contrast sensitivity in the bumblebee Bombus terrestris]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.28.747804v1?rss=1">
<title>
<![CDATA[
Temporal, genome-scale analysis of Myxococcus xanthus developmental fate in a mixed population 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.28.747804v1?rss=1
</link>
<description><![CDATA[
Myxococcus xanthus bacteria form aggregates when starved on solid surfaces and some cells differentiate into spores. Studies of mutants in monoculture have advanced knowledge of this multi-cellular developmental process, but our understanding of the genetic determinants is incomplete. To assess gene function genomewide, we generated a pool of barcoded transposon insertion mutants, subjected it to starvation, and separated developmental samples into non-aggregated cells, aggregated cells, and spores. We also subjected our pool to chemically-induced unicellular sporulation. Evaluation of changes in the abundance of mutants in samples allowed identification of 200 genes in which insertions reproducibly caused distinct patterns of depletion and/or accumulation over time. Many of these genes have well-established roles in development, validating our approach, while many others have not previously been associated with development. Genes involved in type IV pili (T4P)-dependent motility were more important than gliding motility genes for aggregation and sporulation in the mixed population. Although exopolysaccharide (EPS) synthesis genes are required for aggregation in monoculture, most were dispensable for aggregation in our pool, consistent with EPS sharing between cells, yet these genes were required cell-autonomously for efficient sporulation. Genes for positive regulators of EPS synthesis were important for aggregation as well as sporulation, suggesting functions beyond EPS production. Insertions in several novel genes impaired both starvation- and chemically-induced sporulation. Many genes increased the efficiency of starvation-induced sporulation. Some of these mutants, which we call "developmental winners", are novel cheaters. Our results demonstrate the power of using the newly-created mutant library to elucidate M. xanthus biology.
]]></description>
<dc:creator><![CDATA[ Mittal, S., Mandal, S., Farrugia, M. A., Crosson, S., Fiebig, A., Kroos, L. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.28.747804</dc:identifier>
<dc:title><![CDATA[Temporal, genome-scale analysis of Myxococcus xanthus developmental fate in a mixed population]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748126v1?rss=1">
<title>
<![CDATA[
Genome-scale label-free imaging reveals cellular physiology encoded in bacterial collective architecture 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748126v1?rss=1
</link>
<description><![CDATA[
DNA sequencing unified microbial genotyping into a single, comprehensive readout, yet phenotyping remains a slow and fragmented endeavor. Here, we introduce Microbial Phenotyping Using Low-magnification Label-free Imaging (PULLI), a computer vision platform that extracts microcolony and population-level phenotypes from brightfield timelapses of liquid culture growth. Using PULLI, we screened a genome-scale Vibrio cholerae mutant library, recording more than 200,000 images, which revealed that core bacterial pathways shape community architecture. Functionally related mutants converge in appearance, allowing us to resolve processes as distinct as biofilm formation, motility, central metabolism, cofactor biosynthesis, and envelope composition using a single approach. We further show PULLI can be used to determine a drug target, characterize other pathogens, and classify bacterial species. Our results show that bacterial multicellular development is an interpretable signature of genotype-phenotype relationships, which can be captured from simple brightfield timelapses. We release the PULLI pipeline and an interactive atlas of community forms.
]]></description>
<dc:creator><![CDATA[ Mellick, S. N. S., Derringer, J. J., Boyes, D., Croteau, G., Burke, M., Gifford, S., Stark, D. J., Mike, L. A., Turecki, S., Carja, O., Mikheyeva-Bridges, I. V., Bridges, D. A. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748126</dc:identifier>
<dc:title><![CDATA[Genome-scale label-free imaging reveals cellular physiology encoded in bacterial collective architecture]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.31.748290v1?rss=1">
<title>
<![CDATA[
Soil Microbial and Biochemical Properties under Conservation Agriculture in rice-based cropping systems in lower Indo-Gangetic Plain of West Bengal 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.31.748290v1?rss=1
</link>
<description><![CDATA[
Microbial and biochemical properties of soil respond quickly with management practices, than chemical and physical properties. Moreover, impact of conservation agriculture (CA) on soil microbial properties is limited to microbial enumeration, but its effect on soil enzyme and microbial activity is little documented. To address these problems soil enzyme activities [dehydrogenase (DHA), {beta}-glucosidase (BGA), acid phosphatase (AcP) and alkaline phosphatase (AlP) and fluoresceine diacetate (FDA)], microbial activites ((Nitrogen fixation (NFBAct), Phosphate solubilization (PSBAct) & Cellulolytic activities (CDBAct)), microbial biomass ((Soil microbial biomass carbon (SMBC) & soil microbial biomass nitrogen (SMBN)) and available nutrient were studied to evaluate biological soil health in alluvial soil of lower Indo-Gangetic plain (IGP) under CA. Field experiment was conducted in split plot design (SPD), under 3 cropping systems (RMCp: rice-maize-cowpea; RWGg: rice-wheat- green gram; RCfBr; rice-cauliflower- bororice/summer rice). Tillage operations (CT: conventional; MT: minimum and ZT: zero tillage) was main plot and residue application as sub plot treatments [(R0 (no residue), R50 (50% residue) and R100 (100% residue)], treatments were replicated thrice. Biological soil health index (BSHI) indicated that among different degree of CA, ZT (0.464) and (MT=0.441) and R100 (0.464) treatment showed better response. Among different cropping system RMCp (0.359) & RWGg (0.343) outperformed RCfBr (0.609) cropping system with respect to (wrt) microbial and biochemical properties of the soil. Results indicated that for restoring microbial and biochemical properties of soil CA can be used as sustainable practice to restore agro-ecosystem. Keywords: Conservation agriculture, Cropping systems, Soil enzyme, Soil microbial properties, Residue application, Tillage operations.
]]></description>
<dc:creator><![CDATA[ Singh, P., Jaison, M., Saha, N., Dutta, S., Sen, A., Biswas, T., Mandal, B., Mukherjee, S., Dash, B., Sahu, B., Patel, R., Dasgupta, A. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.31.748290</dc:identifier>
<dc:title><![CDATA[Soil Microbial and Biochemical Properties under Conservation Agriculture in rice-based cropping systems in lower Indo-Gangetic Plain of West Bengal]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.31.748220v1?rss=1">
<title>
<![CDATA[
Contrasting evolutionary trajectories of nitrate assimilation across Brettanomyces bruxellensis lineages 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.31.748220v1?rss=1
</link>
<description><![CDATA[
Brettanomyces bruxellensis is a yeast species associated with diverse fermentation environments and characterized by extensive genetic diversity, including diploid, autotriploid, and allotriploid lineages resulting from independent hybridization events. These lineages are associated with distinct ecological niches and provide a framework for studying metabolic trait evolution in complex genomes. Nitrate assimilation is a relatively uncommon trait among yeasts and has been reported in B. bruxellensis, but its distribution and evolutionary history within the species remain poorly understood. Here, we combined phenotypic characterization of 151 strains with genomic analyses of 946 whole-genome sequences to investigate nitrate assimilation. Growth assays revealed that nitrate assimilation is widespread but unevenly distributed across genetic lineages, with some populations largely retaining the trait whereas others have frequently lost it. Genomic analyses identified extensive variation affecting the nitrate assimilation gene cluster composed of YNR1, YNI1, and YNT1. Nitrate assimilation was strongly associated with both gene copy number and predicted gene functionality, with nitrate-assimilating strains generally carrying more functional copies of the cluster. Leveraging the complex genomic architecture of the species, we independently analyzed primary and acquired genomes in allotriploid lineages and uncovered contrasting evolutionary trajectories following hybridization. While nitrate assimilation genes were generally maintained in primary genomes, acquired genomes showed a higher prevalence of gene loss and predicted loss-of-function variants, revealing asymmetric dynamics between subgenomes. Altogether, our results suggest that nitrate assimilation represents an ancestral trait that has been differentially maintained across B. bruxellensis lineages through a combination of copy number variation, gene degeneration, and genome-specific evolutionary dynamics. These findings provide new insights into how genome architecture and polyploid evolution shape the maintenance and loss of metabolic traits in an industrially relevant yeast species.
]]></description>
<dc:creator><![CDATA[ Vigna, A., Harrouard, J., Miot-Sertier, C., Loegler, V., Marullo, P., Friedrich, A., Schacherer, J., Peltier, E., Albertin, W. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.31.748220</dc:identifier>
<dc:title><![CDATA[Contrasting evolutionary trajectories of nitrate assimilation across Brettanomyces bruxellensis lineages]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748096v1?rss=1">
<title>
<![CDATA[
Rapid isothermal amplification of diatom rbcL from eDNA and eRNA reveals their abundance and photosynthetic physiology 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748096v1?rss=1
</link>
<description><![CDATA[
Diatoms are major contributors to marine primary production, yet current approaches for monitoring their abundance and function rely on coarse satellite chlorophyll estimates or sparse cell count and carbon fixation measurements. Molecular markers are a promising approach for high-resolution measurement of both abundance and metabolic activity through analysis of environmental DNA (eDNA) and RNA (eRNA). We present an isothermal quantitative recombinase polymerase amplification (qRPA) assay targeting rbcL gene copies and transcripts of marine diatoms, operating at low temperature and producing results in less than 15 min. We demonstrate specificity and calibration across diverse diatom taxa, then apply the assay to eDNA and eRNA samples from the Mare Chiara Long-Term Ecological Research site in the Bay of Naples, Italy, alongside microscopy, chlorophyll, physicochemical, and carbon-fixation data. Diatom rbcL DNA tracked abundance across five orders of magnitude despite seasonal shifts in community composition. Combining molecular and optical data revealed increased cellular rbcL copies and chlorophyll in low-light winter populations, suggesting enhanced photosynthetic capacity despite lower abundance. Furthermore, rbcL RNA reflected total carbon fixation rates and identified populations with differing carbon fixation activity. These results support rapid, RPA-based rbcL quantification as a robust approach for biomolecular ocean observing.
]]></description>
<dc:creator><![CDATA[ Verret, F. G., Hartle-Mougiou, K., Chantzaras, C., Peltekis, A., Margiotta, F., Sarno, D., Cardini, U., Alba, M., Pizziol, V., Markopoulos, I., Papadopoulou, I., Percopo, I., Tramontano, F., Maselli, M., Novellino, A., Psarra, S., Montresor, M., Mowlem, M. C., Gizeli, E., Valiadi, M. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748096</dc:identifier>
<dc:title><![CDATA[Rapid isothermal amplification of diatom rbcL from eDNA and eRNA reveals their abundance and photosynthetic physiology]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748135v1?rss=1">
<title>
<![CDATA[
RNA virus infection reshapes carbon and nitrogen partitioning in a marine diatom. 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748135v1?rss=1
</link>
<description><![CDATA[
Viral infection is a major yet poorly quantified driver of microbial interactions and biogeochemical fluxes in the ocean. In diatoms, which are key contributors to marine primary production, the extent to which viruses reprogram host cell metabolism and alter elemental cycling remains largely unresolved. Here, we investigated how infection by a lytic single-stranded RNA virus reshapes carbon (C) and nitrogen (N) fluxes in the ecologically relevant nanoplanktonic diatom Mediolabrus comicus. Using controlled infection experiments coupled with flow cytometry, electron microscopy, PAM fluorimetry, and stable isotope probing, we resolved infection-driven changes from the population to the cellular scale. Infection induced rapid optical shifts and cellular reorganization, including the formation of membrane-bound viral replication compartments. These changes coincided with early impairment of plastidial functions, as shown by disruption of photosystem II functionality and a concomitant decline in photosynthetic carbon fixation. In contrast, nitrogen uptake was maintained and strongly enhanced during late stages of infection, indicating sustained resource acquisition to support viral replication. This decoupling led to dynamic changes in cellular stoichiometry and, overall, to substantial reductions in population-level carbon and nitrogen assimilation due to growth inhibition. Together, these findings demonstrate that diatom RNA virus infection reshapes host carbon and nitrogen metabolism, with cascading effects on elemental cycling. Our results identify diatom RNA viruses as important drivers of marine biogeochemical processes, with implications for primary production and the fate of organic matter in the ocean.
]]></description>
<dc:creator><![CDATA[ Jaouen, E., Fiorile, C., Riera, P., Blondel, L., Gachenot, M., Le Gall, F., Nogaret, P., Leroux, C., Six, C., Le Panse, S., Probert, I., Gourvil, P., Bigeard, E., Simon, N., Baudoux, A.-C. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748135</dc:identifier>
<dc:title><![CDATA[RNA virus infection reshapes carbon and nitrogen partitioning in a marine diatom.]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748090v1?rss=1">
<title>
<![CDATA[
Defining the role of aerobic respiration in the metabolism and bioenergetics of Enterococcus faecalis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748090v1?rss=1
</link>
<description><![CDATA[
Enterococcus faecalis is an opportunistic pathogen and facultative anaerobe that primarily relies on fermentative metabolism to colonize a wide range of aerobic and anaerobic environments. In the presence of exogenous heme, E. faecalis can assemble a minimal electron transport chain consisting of membrane-associated primary dehydrogenases, demethylmenaquinone, and the terminal cytochrome bd oxidase (CydAB). This respiratory chain is thought to generate a proton motive force to drive ATP synthesis via the F-type ATP synthase, thereby improving energy conservation under aerobic conditions. However, a cytosolic NADH oxidase (Nox) also consumes NADH and oxygen, potentially competing with the electron transport chain for reducing equivalents and terminal electron acceptors; but the relative physiological contributions of these two oxygen-reducing pathways remain poorly understood. To define the roles of CydAB and Nox under normoxic and hypoxic conditions, we constructed {Delta}cydAB and {Delta}nox mutants. Real-time, in situ measurements revealed {Delta}cydAB had no significant effect on oxygen utilization while in the {Delta}nox it was significantly reduced; revealing Nox as the major consumer of oxygen. Semi-untargeted metabolomic analysis further revealed oxidase-specific alterations in central metabolism with the {Delta}nox causing pronounced shifts in the ATP and NADH ratios; highlighting Nox as a key determinant of intracellular redox and energy homeostasis. Finally, single-cell fluorescence microscopy showed that membrane potential, a component of proton motive force, was substantially diminished only in the absence of both CydAB and Nox, or the F-type ATP synthase. These findings indicate that the F-type ATP synthase is a major generator of proton motive force, even upon aerobic growth, and demonstrate a complementary role for the electron transport chain and Nox in the bioenergetics of E. faecalis.
]]></description>
<dc:creator><![CDATA[ Paxie, O., Nijagal, B., Todd Rose, F. O., Gastrell, S., Su, S., Saleh, A., Grimshaw, J. W., Rhee, K., Strahl, H., Cook, G. M., Darnell, R. L. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748090</dc:identifier>
<dc:title><![CDATA[Defining the role of aerobic respiration in the metabolism and bioenergetics of Enterococcus faecalis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.747672v1?rss=1">
<title>
<![CDATA[
Effect of Mushroom-Bacteria Co-culture on Mushroom Growth and Antimicrobial Properties 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.747672v1?rss=1
</link>
<description><![CDATA[
Edible mushrooms have been reported to have antimicrobial properties and other health benefits. This study aims to test the antimicrobial activities of several edible mushrooms from markets and test if co-culturing them with bacteria could induce stronger anti-bacterial properties. Commercial mushrooms, Hericium erinaceus (lions mane), Pleurotus ostreatus (oyster mushroom), Lentinula edodes (Shiitake) and Agaricus bisporus (button mushroom), were grown from strictly controlled/sterile substrates. Ethanol and water extracts from the mushrooms were prepared and tested against the bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Bacillus subtilis, and the fungus Candida albicans for antimicrobial activities. Shiitake water extract (SWE) showed strong antibacterial effects against all tested bacterial species, inhibitory effects on their biofilms, and antifungal activity. The antimicrobials in SWE seem to damage the cell wall and cell membrane of the bacteria, prefer weak acidic conditions, and are heat labile. Some antimicrobials are likely proteins and polysaccharides. In contrast, 3 other mushrooms displayed only weak antimicrobial effects. The fast-growing lions mane and oyster mushroom were co-cultured with different bacteria. The co-cultivation promoted the fruiting body development of lions mane. Co-culturing with S. aureus increased the anti-bacterial effects of lions mane against S. aureus, E. coli and particularly B. subtilis. Co-culturing the oyster mushroom with bacteria, especially B. subtilis and P. aeruginosa, boosted the mushroom growth. All tested bacteria, especially S. aureus, increased oyster mushroom anti-bacterial effect against E. coli and B. subtilis. The findings indicate that mushroom-bacteria co-culturing could have benefits both agriculturally and medicinally.
]]></description>
<dc:creator><![CDATA[ Wang, E., Cavanaugh, N. T., He, Y., Chai, Y. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.747672</dc:identifier>
<dc:title><![CDATA[Effect of Mushroom-Bacteria Co-culture on Mushroom Growth and Antimicrobial Properties]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748174v1?rss=1">
<title>
<![CDATA[
Receptor-binding domain 2 of Clostridioides difficile binary toxin as a promising vaccine component against C. difficile infection 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748174v1?rss=1
</link>
<description><![CDATA[
Symptoms of Clostridioides difficile infection (CDI) are primarily caused by two major protein toxins, toxin A (TcdA) and toxin B (TcdB). In addition, approximately 5-30% of C. dif[fi]cile strains produce a third toxin, C. difficile binary toxin (CDT), which is has been associated with enhanced virulence and severe disease. CDT consists of an enzymatic component CDTa, and a binding and translocation component CDTb, which mediates the delivery of CDTa into host cells. CDTb contains two receptor-binding domains, RBD1 and RBD2. Recent structural studies suggest that RBD2 plays a critical role in the formation and stabilization of the di-heptameric CDTb assembly required for efficient intoxication of host cells. In this study, we evaluated the immunogenicity and protective potential of RBD1 and RBD2 using in silico, in vitro and in vivo approaches. Sequence analysis demonstrated that RBD2 is highly conserved among diverse CDT-producing C. difficile ribotypes and toxinotypes. Immunization of mice with RBD2, but not RBD1 conferred effective protection against direct CDT challenge. Moreover, RBD2 immunization protected hamsters against infection with a CDT-only-producing C. difficile strain (DSM 101085; TcdA-TcdB-CDT). Mechanistically, anti-RBD2 serum, but not anti-RBD1 serum, effectively neutralized CDT-mediated cytotoxicity, as demonstrated by inhibition of cell rounding in Vero cells. Collectively, these findings identify RBD2 as a promising vaccine antigen targeting CDT and provide functional evidence supporting its critical role in CDT-mediated host-cell intoxication. Incorporation of RBD2 into multivalent C. difficile vaccines may broaden protection against hypervirulent, CDT-producing strains.
]]></description>
<dc:creator><![CDATA[ Wang, S., Heuler, J. S., Nakanishi, Y., Kim, H. B., Sun, X. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748174</dc:identifier>
<dc:title><![CDATA[Receptor-binding domain 2 of Clostridioides difficile binary toxin as a promising vaccine component against C. difficile infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.747438v1?rss=1">
<title>
<![CDATA[
Osmotic adaptation rather than stress response: A time-resolved proteomic analysis of PEG-induced water limitation in Phytophthora cinnamomi 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.747438v1?rss=1
</link>
<description><![CDATA[
Water availability is critical for plants and their microbial communities, including pathogens. The plant pathogen Phytophthora cinnamomi persists in soils with fluctuating moisture, yet cellular responses to water limitation remain poorly understood in Phytophthora and oomycetes more broadly. Although we recently characterized the proteomic response of P. cinnamomi to NaCl-induced osmotic and ionic stress, its response to PEG-mediated water limitation remains poorly understood, leaving a critical gap in our understanding of drought-relevant stress adaptation. Here, we quantified mycelial growth and profiled time-resolved proteome dynamics of P. cinnamomi during polyethylene glycol (PEG-3350)-treatment, simulating moderate water limiting conditions. Treatment with 5% PEG-3350 enhanced radial mycelial growth relative to controls, with no early growth inhibition observed. Label-free proteomics identified 1,097 protein groups, with 880 proteins shared between conditions and an asymmetric abundance profile dominated by decreasing protein abundance over time. Only a small subset of proteins increased, mainly enzymes involved in redox buffering (e.g., thioredoxin and glutaredoxin-like proteins) and mitochondrial/metabolic regulation (e.g., alternative oxidase) and mitochondrial/metabolic regulation. Hierarchical clustering revealed a potential three-phase temporal program: early translational and regulatory remodeling (1-6 HPT), sustained metabolic adjustment (6-12 HPT), and delayed engagement of redox and proteostasis functions (12-24 HPT). Network analysis demonstrated that redox-associated function was integrated throughout this adaptation, with individual clusters further specialized by cofactor preference (NADP- versus NAD-dependent enzymes) and distinct metabolic roles (malate dehydrogenase, CoA-ligase activity). This coordinated, multi-phase reorganization sustained mycelial growth despite moderate osmotic stress, indicating that P. cinnamomi employs active proteomic adaptation rather than passive stress tolerance. These findings reveal the cellular mechanisms underlying drought persistence in this invasive pathogen and suggest molecular targets for disease management under water-limited conditions.
]]></description>
<dc:creator><![CDATA[ Vinson, L. S., Loo, T., Kulshreshtha, S., Dobson, R. C. J., Meisrimler, C. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.747438</dc:identifier>
<dc:title><![CDATA[Osmotic adaptation rather than stress response: A time-resolved proteomic analysis of PEG-induced water limitation in Phytophthora cinnamomi]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748177v1?rss=1">
<title>
<![CDATA[
Differential Biofilm Susceptibility and Potent Isavuconazole Post-Antifungal Effect Distinguish Cutaneotrichosporon dermatis from Trichosporon asahii 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748177v1?rss=1
</link>
<description><![CDATA[
Cutaneotrichosporon dermatis (formerly Trichosporon dermatis) is a basidiomycetous yeast-like fungus known to cause summer-type hypersensitivity pneumonitis, although its virulence in humans remains poorly understood. We performed morphological and molecular identification of an isolate from the sputum and blood cultures of an immunocompromised patient, together with pathogenicity assessment using a Galleria mellonella model, biofilm formation/eradication assays, antifungal susceptibility testing, drug combination effects, and the post-antifungal effect (PAFE), compared with Trichosporon asahii. The isolate was identified as C. dermatis by ITS/IGS1 sequencing, supported by phylogenetic analysis. Growth of C. dermatis increased more at 37 than at 25. In the Galleria mellonella assay, C. dermatis, T. asahii, and Candida albicans each showed dose-dependent pathogenicity at sufficiently high inocula, although Rhizopus oryzae was the most potent pathogen on a per-CFU basis. C. dermatis formed biofilms that were more completely inhibited by terbinafine (TRB) and amphotericin B (AmB) than azole agents, which showed only partial inhibitory activity even at high concentrations. Susceptibility testing showed relatively strong susceptibility to AmB and azole agents. In the TRB and azole combination assay, the fractional inhibitory concentration index (FICI) was below 0.5, indicating synergy. Isavuconazole (ISC) showed a markedly stronger PAFE than the other azole agents tested. These findings indicate that although azoles show only partial efficacy against its biofilm, C. dermatis can still cause invasive infection, and that azole monotherapy or TRB and azole combination therapy, aided by the potent PAFE of ISC, may represent effective treatment options.
]]></description>
<dc:creator><![CDATA[ Yoshinouchi, T., Nakamura, T., Mori, D., Yasunaga, J.-i., Tanaka, Y. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748177</dc:identifier>
<dc:title><![CDATA[Differential Biofilm Susceptibility and Potent Isavuconazole Post-Antifungal Effect Distinguish Cutaneotrichosporon dermatis from Trichosporon asahii]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748148v1?rss=1">
<title>
<![CDATA[
The RNA virome of early metazoans sheds light on long-term virus-host relationships 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748148v1?rss=1
</link>
<description><![CDATA[
Ctenophores and placozoans arose early in metazoan evolution and are characterized by traits associated with key aspects of animal evolution. Despite the evolutionary significance of ctenophores and placozoans, their RNA viromes are poorly understood. To determine the diversity and evolution of RNA virome in these organisms, particularly whether the viruses present with these ancient host lineages similarly occupy basal phylogenetic positions, we analysed publicly available transcriptome data from the Sequence Read Archive (SRA). Accordingly, we identified 26 putative novel viruses classified into 11 virus groups, including members of the families Flaviviridae and Chuviridae. The novel viruses clustered with those previously identified in vertebrates, invertebrates, plants and fungi. Notably, some virus sequences within the Flaviviridae, Chuviridae, Lispiviridae and Marnaviridae were highly divergent, branching deeply relative to their closest known relatives or forming distinct lineages, in some cases suggesting a divergence early in metazoan evolution. In contrast, viruses within the Birnaviridae, Endornaviridae, Mymonaviridae, Narnaviridae, Phasmaviridae, Orthomyxoviridae, Orthototiviridae, and some viruses within the Picornavirales, exhibited patterns consistent with more recent diversification and host jumping. In addition, RNA viruses were detected across multiple species and tissues within the Ctenophora (including whole organisms and embryos) and Placozoa, expanding their host range and highlighting a largely uncharacterized diversity. Together, these findings expand the known diversity and host range of several virus groups, and shed light on virus evolution in early metazoans, demonstrating both host jumping within aquatic environments and virus host-associations that may span the entirety of animal evolution.
]]></description>
<dc:creator><![CDATA[ Ortiz-Baez, A. S., Mifsud, J. C. O., Schwarz, J., Sadiq, S., Holmes, E. C. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748148</dc:identifier>
<dc:title><![CDATA[The RNA virome of early metazoans sheds light on long-term virus-host relationships]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747087v1?rss=1">
<title>
<![CDATA[
Design and characterization of broadly protective influenza A(H3N2) vaccine candidates using protein language models 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747087v1?rss=1
</link>
<description><![CDATA[
Seasonal influenza A viruses cause significant global morbidity each year. Although vaccination remains the primary preventive strategy, effectiveness is often reduced by antigenic drift. This challenge is particularly pronounced for influenza A(H3N2), which has required eight vaccine updates over the past decade. Here, we present a computational framework to engineer broadly reactive influenza A(H3N2) vaccines, using protein language models to generate novel hemagglutinin (HA) sequences and a machine learning model to predict antigenic distance from circulating strains. In a proof-of-concept study, seven HA candidates designed using sequence data from 2013-2018 were evaluated in mice against contemporary and subsequently circulating viruses. Two candidates elicited protective levels of reactive antibodies, robust H3-specific antibody-secreting cell responses, and cross-neutralization against contemporary clades and drifted 2019-2020 strains. These findings demonstrate that an integrated generation-selection strategy can enhance vaccine coverage across current and future A(H3N2) seasons and may be applicable to other influenza subtypes.
]]></description>
<dc:creator><![CDATA[ Howard, V. R., Allen, J. D., Thomas, M. H., Sautto, G. A., Ross, T. M., Georgiev, I. S. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747087</dc:identifier>
<dc:title><![CDATA[Design and characterization of broadly protective influenza A(H3N2) vaccine candidates using protein language models]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.745682v1?rss=1">
<title>
<![CDATA[
Maturation-dependent splicing alterations constrain SYNGAP1 splice-switching therapy 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.745682v1?rss=1
</link>
<description><![CDATA[
Haploinsufficiency in SYNGAP1 causes a severe neurodevelopmental syndrome. SYNGAP1 protein is mainly detected in neuronal synapses. However, SYNGAP1 RNA is more widely expressed and strongly regulated via alternative splicing: alternative 3' splice site (A3SS) inclusion leads to non-productive transcripts that are degraded through nonsense-mediated decay. Recently, splice-switching oligonucleotides (SSOs) that redirect SYNGAP1 splicing to increase SYNGAP1 protein levels were developed. However, we hypothesized that during neuronal maturation, non-productive splicing may decrease to enhance functional transcripts in mature neurons. This would reduce the abundance of the SSO target transcript, limiting the potential for SSO treatment to increase neuronal SYNGAP1 expression. Using neural differentiation of human induced pluripotent stem cells, we show that the A3SS transcript is abundant in neural progenitors, astrocytes, microglia and immature neurons, with minimal presence in mature neurons. These data imply that SSOs targeting A3SS might lack therapeutic efficacy to rescue the neuronal phenotypes associated with SYNGAP1 haploinsufficiency.
]]></description>
<dc:creator><![CDATA[ Kamp, J. A., Wijnant, K. A., Maas, N., Gülyurt, D., Rieder, M. J., Jolfaei, M. A., Gontan, C., Kushner, S. A., Elgersma, Y., Vissers, L. E., Nadif Kasri, N., De Vrij, F. M. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.745682</dc:identifier>
<dc:title><![CDATA[Maturation-dependent splicing alterations constrain SYNGAP1 splice-switching therapy]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747343v1?rss=1">
<title>
<![CDATA[
Early MATR3 loss and distinct neurodegenerative molecular signatures precede the onset of neuropathology in motor neurons and Purkinje cells of MATR3 S85C knock-in mouse model of ALS 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747343v1?rss=1
</link>
<description><![CDATA[
Amyotrophic lateral sclerosis (ALS) is a motor neuron disease, leading to progressive muscle weakness and motor impairment. Growing evidence indicates that cerebellar Purkinje cells, which play a central role in motor coordination, are also affected in ALS. However, it is unclear whether the molecular events that initiate neurodegeneration in these ALS-relevant motor-controlling neurons are shared or distinct. Here, we used a MATR3 S85C knock-in (KI) mouse model of early-stage ALS with stage-specific motor phenotypes and selective vulnerability of motor neurons and Purkinje cells to decipher the molecular events underlying neurodegeneration in these two neuronal populations. We found that a profound reduction in detectable MATR3 S85C immunoreactivity (hereafter referred to as MATR3 loss) in both motor neurons and Purkinje cells precedes the onset of motor dysfunction and neuropathology, implicating MATR3 loss as the earliest detectable molecular event. Our bulk cerebellar RNA profiling and motor neuron-specific RNA profiling data at the onset of MATR3 loss revealed distinct molecular signatures. In the cerebellum, Ngfr expression emerged in Purkinje cells before the onset of neuronal loss and remained elevated throughout the disease course. This increase was accompanied by activation of the JNK-mediated cell death pathway. In the motor neurons, elevated Fgf21 and integrated stress response (ISR) gene expression were the first to be observed and persisted throughout disease progression, consistent with previous findings in SOD1 mouse models. Our findings provide mechanistic insights into the initiation of neurodegeneration in ALS-relevant motor-controlling neurons and implicate potential neuron type-specific targets for future therapeutics.
]]></description>
<dc:creator><![CDATA[ Maksimovic, K., Majji, R., Santos, J. R., Chan, C., Zelaya, A., Lee, J., Dias, M., Gluscencova, O. B., Youssef, M. M. M., Kim, S., Noronha, T., Lai, C., Fan, Y., Metri, M. N., You, J., Kao, C. S., Wang, L.-Y., Lefebvre, J. L., Wilson, M. D., Yalamanchili, H. K., Park, J. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747343</dc:identifier>
<dc:title><![CDATA[Early MATR3 loss and distinct neurodegenerative molecular signatures precede the onset of neuropathology in motor neurons and Purkinje cells of MATR3 S85C knock-in mouse model of ALS]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.25.747102v1?rss=1">
<title>
<![CDATA[
A neuronal CRISPRi screen identifies PQLC2 as a lysosomal pH regulator controlling tau homeostasis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.25.747102v1?rss=1
</link>
<description><![CDATA[
Lysosomes make key contributions to the maintenance of cellular proteostasis, and their functional compromise has been linked to aging and neurodegenerative disease. A defining characteristic of lysosomes is their relative acidity compared to other subcellular compartments, a quality that enables the efficient breakdown of macromolecules. Evidence suggests that neuronal lysosomal pH becomes dysregulated with aging and neurodegenerative disease, yet the mechanisms by which lysosomal pH is maintained remain incompletely understood. To better understand neuronal lysosomal pH regulation, we conducted a genome-wide CRISPRi-based screen in iPSC-derived iNeurons for modifiers of lysosomal pH. We validated several previously known regulators of lysosomal pH and identified novel pathways capable of modifying lysosomal pH, including protein UFMylation and mitochondrial homeostasis. We demonstrate that loss of the lysosomal cationic amino acid exporter, PQLC2, prevents lysosomal acidification in a manner independent of amino acid transport. A novel, tauopathy-associated mutation in PQLC2 impairs lysosomal acidification and drives tau accumulation. Together, this study reveals novel genes that modify lysosomal pH and highlights potential new targets for ameliorating age-related lysosome dysfunction.
]]></description>
<dc:creator><![CDATA[ Welch, M., Sampognaro, P. J., Shu, S., Chaplot, K., Bothra, A., Castruita, P. A., Smith, A. W., Antee, T., Hodul, M., Tian, R., Gao, V., Limas, J. C., Burris, K. D., Parker, J. L., Yokoyama, J. S., Miller, B. L., Seeley, W. W., Newstead, S., Kampmann, M., Kao, A. W. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.25.747102</dc:identifier>
<dc:title><![CDATA[A neuronal CRISPRi screen identifies PQLC2 as a lysosomal pH regulator controlling tau homeostasis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.26.747219v1?rss=1">
<title>
<![CDATA[
Learning a threat converges on the circuit processing innate threat 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.26.747219v1?rss=1
</link>
<description><![CDATA[
A prevailing view in affective neuroscience holds that innate and learned behaviors are processed through distinct neuroanatomical pathways, one pre-wired, the other running on synaptic plasticity. However, here we show that processing innate and learned threats in the lateral amygdala deviates fundamentally from this view. We tracked the three core elements of circuit function (excitatory neurons, inhibitory neurons, and neuromodulators) in mice, as they were exposed to an innately aversive looming stimulus and as they learned a cued threat. Tracking the same neurons across sessions, revealed a subpopulation of excitatory neurons recruited by the innate threat that was preferentially potentiated following auditory threat learning. Furthermore, both forms of threat converged on the same modulatory mechanisms: the disinhibitory VIP/SST motif and norepinephrine release, but with a critical difference. While the innately aversive stimulus possessed privileged access to these pathways, the learned cue acquired access through synaptic plasticity. In this instance, learning about a new threat apparently recruits a circuit that protects animals from natural threats.
]]></description>
<dc:creator><![CDATA[ Mermet-Joret, N., Nazari, M., Pommer, A. T., Ansarifar, S., Silva Luz, J., Vestergaard, A.-K., Nabavi, S. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.26.747219</dc:identifier>
<dc:title><![CDATA[Learning a threat converges on the circuit processing innate threat]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.29.747960v1?rss=1">
<title>
<![CDATA[
Glioblastoma Tumors with Decelerated Epigenetic Aging Are Characterized by Glutamatergic Neuronal Activity and Stemness 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.29.747960v1?rss=1
</link>
<description><![CDATA[
Introduction: Gliomas integrate into neural circuits and heighten neuronal excitability, engaging in bidirectional communication whereby neuronal activity promotes tumor growth and proliferation. Aging reshapes the brain microenvironment through extracellular matrix changes, altered secretory factors, and immune dysfunction, creating conditions permissive to tumorigenesis and limiting immunotherapy efficacy in glioblastoma. However, its effect on neuronal excitability and signaling in glioblastoma remains poorly understood. Methods: We developed a novel classification system for glioblastoma by leveraging three classes of DNA methylation-based aging biomarkers: chronological, biological, and mitotic clocks. This approach stratified tumors into accelerated and decelerated epigenetic aging subtypes, which we then characterized at the molecular, functional, and clinical levels using multimodal analyses. Guided by these profiles, we evaluated the in vitro effects of the FDA-approved agents levetiracetam and riluzole, alone and in combination with temozolomide, on U87MG and A172 cell lines. Specifically, we assessed changes in cell viability, apoptosis, and the expression of marker genes related to stemness, neuronal hyperexcitability, and immunosuppression. Results: Tumors with decelerated epigenetic aging showed expression modules and CpG hypomethylation associated with neuronal activity and stemness, and carried significantly worse prognosis. Single-cell and spatial multi-omics analyses revealed enrichment for neurons and malignant neural stem-like cells in these tumors. They also displayed enhanced intercellular communication, driven predominantly by glutamate signaling across the malignant, neuronal, and immune compartments of the tumor microenvironment. In vitro pharmacological inhibition of glutamatergic signaling with levetiracetam and riluzole reduced cell viability, induced apoptosis, and suppressed expression of stemness, neuronal hyperexcitability, and immunosuppression markers. Both agents potentiated the cytotoxic and apoptotic effects of temozolomide, supporting glutamatergic inhibition as a strategy for improving chemosensitivity. Conclusion: By establishing a framework for decoding glioblastoma heterogeneity through epigenetic aging, we identified the glutamatergic pathway as a clinically actionable vulnerability. Our findings suggest that combining anti-glutamatergic therapies with temozolomide exerts synergistic antitumor effects while mitigating adverse chemotherapy-induced phenotypes, such as increased stemness, neuronal hyperexcitability, and immunosuppression, thereby laying the groundwork for novel therapeutic strategies.
]]></description>
<dc:creator><![CDATA[ Motevasseli, M., Eterafi, M., Alaei, H., Zandi, P., Shajari, N., Tabrzi, M., Safarzadeh, E. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.29.747960</dc:identifier>
<dc:title><![CDATA[Glioblastoma Tumors with Decelerated Epigenetic Aging Are Characterized by Glutamatergic Neuronal Activity and Stemness]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.748071v1?rss=1">
<title>
<![CDATA[
Pan-cancer analysis identifies nine conserved miRNA regulators of tumor cytolytic activity and clinically actionable immune targets 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.748071v1?rss=1
</link>
<description><![CDATA[
Abstract Background: Cytolytic activity (CYT), a widely used transcriptomic surrogate of anti-tumor immune cytotoxicity derived from GZMA (granzyme A) and PRF1 (Perforin 1) expression, is associated with clinical outcomes across cancers. MicroRNAs (miRNAs) are key post-transcriptional regulators of tumor immunity, yet their pan-cancer roles in modulating cytolytic activity remain incompletely understood. Objective: This study aimed to identify conserved miRNA regulators of tumor cytolytic activity and their downstream gene-mediated networks across diverse cancer types, while evaluating their clinical and therapeutic relevance. Methods: Matched miRNA and mRNA expression profiles from 9,288 primary tumors across 31 TCGA cancer types were analyzed. A multi-stage framework was applied: per-cancer Spearman correlations (|{rho}| >= 0.30, FDR < 0.05) identified recurrent CYT-associated miRNAs (at least 3 cancer types); these were integrated with TargetScan-predicted targets and subjected to pan-cancer and cross-cancer triple filtering (miRNA-gene and gene-CYT associations). All associations underwent tumor purity adjustment using Consensus Purity Estimate (CPE), with LUMP (Leukocytes Unmethylation for Purity) as sensitivity analysis. Candidates were further prioritized by random forest modeling with bootstrap stability, cancer-type-adjusted Cox regression, mediation analysis, immune cell deconvolution, k-means molecular subtyping, pathway enrichment, and DGIdb-based drug-target prioritization. Results: The analysis converged on 38 high-confidence miRNA-gene-CYT regulatory triplets involving 9 conserved miRNAs and 31 target genes after stringent purity adjustment and multi-layer validation. All nine miRNAs exhibited complete bootstrap stability. Mediation analysis confirmed significant gene-level mediation in 37 of 38 triplets (FDR < 0.01), with mediated proportions up to 94%. The final miRNA signature defined two distinct pan-cancer immune subtypes (immune-hot vs. immune-cold) with significantly different cytolytic activity and overall survival (OS) (HR = 0.754, FDR = 1.12 x 10^-4). The network was enriched for T-cell activation and lymphocyte differentiation pathways and highlighted multiple druggable targets, including CTLA4 and CD274 (PD-L1), nominating 124 candidate compounds. Conclusions: In conclusion, this tumor purity-adjusted pan-cancer study defines a compact, reproducible, and clinically relevant miRNA network that regulates cytolytic activity across diverse malignancies. By linking miRNA biology to immune subtyping and actionable therapeutic targets, the present work provides a valuable foundation for advancing precision immuno-oncology.
]]></description>
<dc:creator><![CDATA[ Bagherlou, N., Aliyari, S., Salehi, Z., Pirouzkhah, M., Weis, C.-A. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.748071</dc:identifier>
<dc:title><![CDATA[Pan-cancer analysis identifies nine conserved miRNA regulators of tumor cytolytic activity and clinically actionable immune targets]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.08.30.747968v1?rss=1">
<title>
<![CDATA[
Cryo-EM structure of CYP2C9 reveals a dimer-of-trimers assembly 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.08.30.747968v1?rss=1
</link>
<description><![CDATA[
Human cytochrome P450 2C9 (CYP2C9) is a hepatic microsomal enzyme involved in the oxidative metabolism of clinically important drugs, but the structural organization of its oligomeric assemblies outside crystallographic packing environments remains poorly understood. Here, we report the cryo-EM structure of human CYP2C9 determined under aqueous, membrane-free conditions at 3.31 Angstrom resolution. The structure reveals a C2-symmetric hexameric assembly organized as a dimer of trimers. Individual protomers retain the conserved P450 fold and heme-binding architecture observed in previously reported crystal structures, indicating that assembly formation does not substantially perturb the catalytic core. The hexamer is stabilized by defined intra-trimer interfaces involving the N-terminal region and residues around Trp212 and Phe482, together with inter-trimer interfaces involving Leu71 and the 220-227 loop. These interfaces are distinct from the crystal packing contacts observed in CYP2C9 crystal structures, demonstrating that the assembly is not a simple recapitulation of crystallographic packing. Notably, the inter-trimer interface is located near the FG-loop-containing surface previously implicated in membrane association. This suggests that the observed hexamer may represent a membrane-free association of two trimers through membrane-related surfaces, whereas the trimeric arrangement itself may be compatible with membrane-associated organization. The structure therefore provides a framework for investigating how trimer formation, membrane interaction and local conformational changes in the FG-loop region may influence CYP2C9 function.
]]></description>
<dc:creator><![CDATA[ Tanino, H., Tsujino, H., Nakao, T., Oie, C., Makino, F., Miyata, T., Kasai, K., Namba, K., Inoue, T. ]]></dc:creator>
<dc:date>2026-08-31</dc:date>
<dc:identifier>doi:10.64898/2026.08.30.747968</dc:identifier>
<dc:title><![CDATA[Cryo-EM structure of CYP2C9 reveals a dimer-of-trimers assembly]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-08-31</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
