mBio最新文献

{"title":"\"Showing the data\" in published biology research.","authors":"Benjamin G Freeman, Marina E Haldopoulos, Sidharth Srinivasan, Taylor L Cooper, Soobin An, Miharu Yasumuro, Aiswarya Venkata Suresh Kumar, Melody Modarressi, Xinyu Zhang, Akul Chopra","doi":"10.1128/mbio.00572-26","DOIUrl":"https://doi.org/10.1128/mbio.00572-26","url":null,"abstract":"<p><p>Showing the data is the first rule of effective figures, yet this mandate is often ignored. Perhaps the signature offender is the \"dynamite plot\"-a bar graph showing mean and error. Here, we evaluate recent trends in the use of dynamite plots by analyzing 8,834 figures from 2,930 studies published between 2021 and 2025 in 18 journals from five fields of biology. We find that dynamite plots constitute ~25% of figures and are especially common in microbiology journals. However, the use of dynamite plots has declined substantially across fields and journals from 30% of figures in 2021 to 18% in 2025-evidence that biologists increasingly show the data in their figures. We advocate for authors, reviewers, and editors to continue this trend, suggest simple dot plots as an effective replacement for dynamite plots, and describe other options when space or sample size makes dot plots less feasible.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0057226"},"PeriodicalIF":4.7,"publicationDate":"2026-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inverse stable isotope labeling (InverSIL) links predicted catecholate siderophore gene clusters to their products in diverse bacteria.","authors":"Jose Miguel D Robes, Tashi C E Liebergesell, Victoria P Medvedeva, Aaron W Puri","doi":"10.1128/mbio.03391-25","DOIUrl":"https://doi.org/10.1128/mbio.03391-25","url":null,"abstract":"<p><p>Bacteria produce high-affinity, iron-chelating secondary metabolites called siderophores to access insoluble Fe(III) in their environments. Genome mining has revealed many predicted siderophore biosynthetic gene clusters (BGCs) in bacterial genomes; however, the structures of their siderophore products remain mostly undetermined. This limits our molecular-level understanding of how bacteria acquire iron. Here, we apply inverse stable isotope labeling (InverSIL) to rapidly connect predicted siderophore BGCs to their products. With InverSIL, bacteria are grown on ¹³C-substituted carbon sources and then fed predicted biosynthetic precursors at their natural isotopic abundance to identify BGC products by mass spectrometry, removing issues with the availability of isotopically substituted precursors. We use InverSIL to determine the structures of the siderophore products of predicted BGCs from the methylotrophic genera <i>Methylophilus</i> and <i>Methylorubrum</i>, as well as the siderophores produced by the opportunistic pathogen <i>Chromobacterium violaceum</i>, which were previously shown to be essential for virulence yet remained structurally uncharacterized. We next use this approach to reveal the unexpected production of enterobactin by the genera <i>Kushneria</i> and <i>Paracoccus</i>, which was difficult to predict from genome sequences due to the distributed nature of the biosynthetic genes within the genomes. Finally, we use InverSIL to discover new siderophores, the cellulochelins, from the cellulose-degrading plant symbiont <i>Cellulomonas</i> sp. strain Leaf334. These findings demonstrate the utility of InverSIL for functional BGC characterization and expand our molecular understanding of bacterial iron acquisition strategies.</p><p><strong>Importance: </strong>Iron acquisition is important for microbial survival, and bacteria produce secondary metabolites called siderophores to scavenge iron from the environment. While bacterial genome sequences show many predicted genes for making siderophores, most remain unlinked to their metabolic products. Understanding which siderophores bacteria produce is critical for elucidating microbial iron acquisition strategies, ecological interactions, and potential roles in host-microbe interactions. Here, we demonstrate how inverse stable isotope labeling (InverSIL) can rapidly link predicted siderophore gene clusters to their corresponding metabolites. By applying InverSIL to diverse bacterial strains, we validate known siderophore products and uncover unexpected products, highlighting the limitations of current <i>in silico</i> predictions. This study highlights the value of combining experimental approaches with genome mining to advance our understanding of how bacteria acquire iron from their environment.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0339125"},"PeriodicalIF":4.7,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839670","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Structural evolution of a fungal cell wall protein family for β-glucan-binding and cell separation.","authors":"Philipp Schöppner, Vitali Weitzel, Maik Veelders, Lukas Korf, Jonas Andräs, Katharina Wolf, Stefan Brückner, Lars-Oliver Essen, Hans-Ulrich Mösch","doi":"10.1128/mbio.03535-25","DOIUrl":"https://doi.org/10.1128/mbio.03535-25","url":null,"abstract":"<p><p>In fungi, the continuous biosynthesis and remodeling of the cell wall are crucial for growth, division, and development. A hallmark of fungal cell walls is their layered structure, which includes several carbohydrate polymers, such as β-glucans, and a large number of associated cell wall proteins. The fungal-specific family of SUN domain proteins has been implicated in cell wall remodeling and cell separation, but detailed structure-based analyses revealing precise molecular functions have been lacking until now. In this study, we determined high-resolution crystal structures of the SUN domains from two paralogs of the SUN family in budding yeast. We find that their bilobal architecture consists of a domain with high structural similarity to the Sushi/SCR/CCP domain (INTERPRO family IPR000436), and an intimately associated thaumatin-like domain (IPR001938). Together, these domains form a highly conserved canyon fitted to accommodate both single- and triple-helical β-glucan polymers. Within this canyon, we identify 12 conserved polar residues that are crucial for the function of SUN domains in mediating cell separation. We further demonstrate that SUN domains are functionally interchangeable between paralogs in budding yeast, as well as between orthologs from budding yeast and phylogenetically distant fission yeast or filamentous fungi. We conclude that the fungal SUN domain family represents a unique class of β-1,3-glucan-binding proteins involved in cell wall remodeling and separation, whose successful evolution was enabled by the fusion of ancestral sushi- and thaumatin-like domains.</p><p><strong>Importance: </strong>Fungal cell walls are dynamic extracellular structures essential for growth and morphogenesis, making them prime targets for antifungal drugs and the host immune system. Although many protein families involved in the synthesis, crosslinking, and degradation of cell wall polymers are known, the molecular functions and structural evolution of most cell wall proteins remain poorly understood. Our in-depth structural, functional, and phylogenetic analysis of the fungal SUN domain protein family sheds light on a central question: how specific protein families have evolved structurally to enable dynamic cell wall remodeling during growth and division. Moreover, this work identifies precise structural targets within the fungal cell wall that could guide the development of novel diagnostics and therapeutics against life-threatening fungal infections.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0353525"},"PeriodicalIF":4.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Loss of nsp14-exonuclease activity impairs the replication, proofreading, fitness, and pathogenesis of SARS-CoV-2.","authors":"Jordan Anderson-Daniels, Meghan V Diefenbacher, Boyd L Yount, Rita M Meganck, Longping V Tse, Kaitlyn N Burke, Hector A Miranda, D Trevor Scobey, Xiaotao Lu, Laura Stevens, Kenneth H Dinnon, Nathaniel S Chapman, Camryn Pajon, John M Powers, Cameron Nguyen, Rachel L Graham, Nicholas S Heaton, Ralph S Baric, Mark R Denison, Timothy P Sheahan","doi":"10.1128/mbio.00073-26","DOIUrl":"https://doi.org/10.1128/mbio.00073-26","url":null,"abstract":"<p><p>Coronaviruses (CoVs) replicate their RNA genomes with a higher degree of fidelity than other RNA viruses, a mechanism mediated by the proofreading and recombination activities of the exoribonuclease domain of replicase nonstructural protein 14 (nsp14-ExoN). Both murine hepatitis virus (MHV) and SARS-CoV tolerate nsp14-ExoN loss-of-function mutations (ExoN-) (D90A and E92A), but have impaired replication fidelity and pathogenesis; yet identical substitutions in MERS-CoV and SARS-CoV-2 have been reported to be lethal. Here, we report a saturation mutagenesis approach facilitating the recovery and analysis of several constellations of SARS-CoV-2 nsp14 ExoN-inactivating, loss-of-function substitutions, including the canonical D90A and E92A. Biochemical assays with purified WT or ExoN-nsp10-14 fusion proteins confirmed that active site substitutions abolished ExoN activity (ExoN-). SARS-CoV-2 ExoN- viruses exhibited impaired replication, RNA synthesis, and recombination, as well as decreased replication fidelity and loss of fitness <i>in vitro</i>. ExoN- viruses were significantly attenuated for replication in human primary airway epithelial cells and were attenuated for replication and pathogenesis in WT mice, as well as the highly susceptible K18 transgenic mice. In the absence of interferon signaling <i>in vivo</i>, SARS-CoV and SARS-CoV-2 ExoN- viral replication could be partially restored. These results demonstrate that SARS-CoV-2 ExoN- viruses are viable but highly impaired for replication, fitness, and fidelity <i>in vitro,</i> as well as innate immune antagonism and pathogenesis <i>in vivo</i>. Collectively, our results solidify the multiple critical roles of nsp14-ExoN across CoV genera and establish new approaches for rescuing and analyzing loss-of-function substitutions in studies of CoV replication, pathogenesis, and evolution.</p><p><strong>Importance: </strong>Coronaviruses (CoV) are important human pathogens causing hundreds of millions of infections and millions of deaths over the past 20 years. The study of how these viruses multiply and cause disease identifies points of attack for therapeutics. Using a high-throughput genetic approach, we systematically inactivated an essential enzyme CoV needs for replication called ExoN. We show that without ExoN, CoV replication fidelity and fitness are reduced in cell culture. Replication without ExoN in mice was diminished but could be partially restored in mice that lack key components of the immune response. Altogether, we reveal new insights into the complexities of CoV replication and virus and host interactions, which could be leveraged for the development of novel multifaceted therapeutics that attack the ever-expanding functions of the CoV replication complex in replication and pathogenesis.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0007326"},"PeriodicalIF":4.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex and regional effects of <i>Bacteroides</i> in the gut.","authors":"Rebecca A Valls, Kaitlyn E Barrack, Sarvesh V Surve, James B Bliska, George A O'Toole","doi":"10.1128/mbio.00469-26","DOIUrl":"https://doi.org/10.1128/mbio.00469-26","url":null,"abstract":"<p><p><i>Bacteroides</i> spp. are a key immune-programming microbe in healthy individuals-these bacteria have been shown to be reduced in abundance across a variety of disease states. Our study investigated the systemic and region-specific responses to <i>Bacteroides</i> colonization in the gut, including sex-related differences, in mice. Utilizing C57BL/6 mice, we administered <i>Bacteroides</i> to conventional, antibiotic-treated mice, then assessed this microbe's influence on the gut microbiota composition and inflammatory responses following an airway lipopolysaccharide challenge to assess effects on the gut-lung axis. We found that <i>Bacteroides</i> successfully colonizes the intestinal tract of antibiotic-treated mice, particularly the colon lumen of the large intestine, as evidenced by 16S rRNA amplicon gene sequencing and culturing. Differential gene expression analysis using NanoString technology revealed significant immune response variations across the gut regions, with notable differences in adaptive immune response genes. A striking sex-dependent outcome was noted in the regulation of <i>atg12</i> in the cecum, potentially enhancing autophagic function, particularly in female mice. Additionally, <i>Bacteroides</i> intestinal colonization was associated with altered expression of macrophage markers such as <i>cd163</i>, <i>cd84</i>, and <i>ms4a4a</i>, which may reflect shifts in the macrophage profile within the cecum. These findings pave the way for novel therapeutic approaches that leverage microbial impacts on gut and systemic health, offering a deeper understanding of <i>Bacteroides'</i> role in human health and disease. Our study highlights the necessity for further research to elucidate the intricate relationships between gut microbiota, host immunity, biological sex, and their interplay.</p><p><strong>Importance: </strong>This research marks an investigation into how specific microbiota, like <i>Bacteroides</i>, regulate host responses across different gut regions to influence systemic health. By dissecting the impact of <i>Bacteroides</i> across multiple regions of the intestinal tract, this study offers new insights into the localized and whole-body effects of this important immune-programming microbe. Such an understanding is crucial as it helps in unraveling the complex interplay between gut microbes and the host's immune system. This research helps bridge the gap between local intestinal ecology and overall systemic health, addresses important questions relevant to the gut-lung axis, and helps pave the way for innovative therapies.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0046926"},"PeriodicalIF":4.7,"publicationDate":"2026-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839867","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transforming the American Academy of Microbiology for social good through scientific portfolios.","authors":"Nguyen K Nguyen, Rachel M Burckhardt, Stefano Bertuzzi, Arturo Casadevall, Vanessa Sperandio, Jay T Lennon","doi":"10.1128/mbio.00828-26","DOIUrl":"https://doi.org/10.1128/mbio.00828-26","url":null,"abstract":"<p><p>As the research landscape evolves, scientific societies must adapt their programs to meet changing community needs. The American Academy of Microbiology (Academy or AAM) has recently developed a new model centered around scientific portfolios aimed at advancing its vision of becoming an effective scientific think tank. Here, we describe this transition and the process used to develop and implement a portfolio-based approach. We highlight the Climate Change and Microbes Scientific Portfolio as a case study, demonstrating its successes and its ability to guide the design of future portfolios.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0082826"},"PeriodicalIF":4.7,"publicationDate":"2026-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-omics profiling of fungal balls in chronic pulmonary aspergillosis patients reveals microbiome dynamics and metabolic adaptations.","authors":"Chan Liu, Matheus Mertz Ribeiro, Jian Yang, Liangyu Li, Jianxiong Li, Xianqiu Chen, Yude Wang, Le-Le Wang, Beibei Wang, Yiming Zhou, Jing Zhang, Jijin Jiang, Jielu Lin, Endrews Delbaje, Jin-Fu Xu, Gustavo H Goldman, Shuo Liang","doi":"10.1128/mbio.00348-26","DOIUrl":"https://doi.org/10.1128/mbio.00348-26","url":null,"abstract":"<p><p>Fungal balls (aspergillomas) are a debilitating complication of chronic pulmonary aspergillosis, but their functional biology as multi-kingdom ecosystems is poorly understood. Through integrated multi-omics analysis of 61 patient-derived fungal balls, we reveal their complex ecology. While <i>Aspergillus fumigatus</i> dominates the fungal niche (59% of patients), bacterial co-colonization is ubiquitous, primarily by <i>Pseudomonas aeruginosa</i> and <i>Haemophilus influenzae</i>. Metabolomics and metatranscriptomics unveil a structured division of labor and active warfare, including metabolic cross-feeding, competition for iron, and reciprocal antagonism via secondary metabolites, such as fumagillin and fumigaclavine C produced by <i>A. fumigatus</i>. Host metabolomics and transcriptomics revealed a potent but dysregulated human immune response, characterized by neutrophil activation and failed resolution. Our findings redefine aspergilloma not as a mere fungal aggregate, but as a resilient polymicrobial biofilm across kingdoms, in which synergistic and antagonistic inter-kingdom interactions drive pathogenesis and chronicity, suggesting new therapeutic strategies targeting the pathogenic consortium.IMPORTANCEChronic pulmonary aspergillosis (CPA) and its hallmark fungal balls (aspergillomas) represent a debilitating and difficult-to-treat respiratory disease, affecting millions worldwide. Here, we provide the first integrated multi-omics profile of surgically resected fungal balls from 61 CPA patients, revealing these structures not as mere fungal colonies, but as resilient, cross-kingdom biofilms teeming with bacterial co-colonizers, particularly <i>Pseudomonas aeruginosa</i> and <i>Haemophilus influenzae</i>. Our findings uncover a dynamic battlefield where fungi and bacteria engage in metabolic cross-feeding, chemical warfare, and competition for nutrients such as iron. We demonstrate that the host mounts a potent but dysregulated immune response characterized by chronic neutrophilic inflammation and failed resolution, driving tissue damage and disease persistence. Our data provide a foundation for novel therapeutic strategies aimed at disrupting microbial synergy, modulating host inflammation, and breaking the cycle of chronic infection, an approach that could significantly improve outcomes for patients with this refractory disease.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0034826"},"PeriodicalIF":4.7,"publicationDate":"2026-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839925","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"mGem: More than building blocks? Mitochondrial metabolism, viruses, and the host response to infection.","authors":"Jessica Alvarez, Dustin C Hancks","doi":"10.1128/mbio.02501-25","DOIUrl":"https://doi.org/10.1128/mbio.02501-25","url":null,"abstract":"<p><p>Beyond essential roles as central hubs integrating homeostatic cellular metabolism, mitochondria have emerged as critical determinants of infection outcomes. Mitochondrial activities, like MAVS signaling and the release of cytochrome c and mitochondrial DNA, drive host defenses. Across cell types, mitochondrial metabolism and antiviral responses are also increasingly being connected by evidence such as viral-encoded antagonists. Nonetheless, metabolic rewiring in infected cells is still largely viewed as a means to satisfy biosynthetic demands for both viral replication and the host response. However, perturbation of metabolic states within infected and bystander cells seemingly has consequences for outcomes, implying an incompletely understood metabo-immunoregulatory logic. Here, we consider roles for mitochondrial metabolism reprogramming as an active cue that licenses progressive immune states to adapt host responses. In the coming years, integration of mitochondrial biology and new methodologies, including spatial approaches, will illuminate the interplay of mitochondrial metabolism on primary antiviral responses.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0250125"},"PeriodicalIF":4.7,"publicationDate":"2026-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Constitutive interferon epsilon expression shapes antiviral epithelial states in the female reproductive tract and intestine.","authors":"Rebecca L Casazza, Samantha Skavicus, David Hare, Kaila A Cooley, Nicholas S Heaton, Carolyn B Coyne","doi":"10.1128/mbio.00340-26","DOIUrl":"https://doi.org/10.1128/mbio.00340-26","url":null,"abstract":"<p><p>Antiviral defenses at mucosal barriers are essential for preventing viral entry and systemic infection. Interferon epsilon (IFNε) is a unique type I IFN that, unlike other family members, is not induced by infection but is constitutively expressed in epithelial tissues. IFNε was initially characterized in the female reproductive tract (FRT), where it provides broad antiviral protection, but its roles outside the FRT remain poorly defined. Here, we used <i>Ifnε^-/-</i> mice and single-cell RNA sequencing to delineate IFNε function across distinct mucosal surfaces. In the FRT, <i>Ifnε</i> expression was restricted to specific epithelial subsets, was independent of estrous stage, and maintained basal ISG expression. IFNε was also retained intracellularly in primary human FRT-derived cells. Extending these analyses to the intestine, we found that IFNε is highly expressed in villous-tip enterocytes of the small intestine <i>in vivo</i>, where it sustains inflammatory enterocyte subsets and maintains type III IFN expression. Loss of <i>Ifnε</i> depleted these subsets and rendered mice more susceptible to enteric viral infection. Together, these findings establish IFNε as a constitutively expressed, spatially restricted IFN that coordinates mucosal antiviral defenses across both reproductive and gastrointestinal epithelial tissues.</p><p><strong>Importance: </strong>Interferon epsilon (IFNε) is a unique type I IFN that, unlike other family members, is not induced by infection but is constitutively expressed in epithelial tissues. In this manuscript, we define the epithelial cell types that constitutively express IFNε in the uterus and small intestine at a single-cell resolution. We show that mice lacking IFNε lose key antiviral defenses in a tissue-dependent manner; uterine epithelial cells have diminished basal ISG expression, and key populations of cytokine-expressing enterocytes are absent from the small intestine. In the intestine, this correlates with increased susceptibility to infection with an enteric virus in mice. These findings establish IFNε as a key contributor to mucosal immunity, sustaining antiviral defenses within tissue-specific epithelial cells of both the female reproductive tract and intestine, and broaden our understanding of its role beyond traditional pathogen-induced interferon responses.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0034026"},"PeriodicalIF":4.7,"publicationDate":"2026-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147839667","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Japanese encephalitis virus NS1 and NS4B synergistically target TLR3 signaling to promote viral replication.","authors":"Quan Zeng, Maozhou He, Mengyao Li, Dong Yang, Lei Wang, Chenlin Hao, Zhaoqing Wang, Minmin Zhou, Tianrenzheng Zhu, Xueshi Niu, Zhihui Chen, Bingqian Zhang, Shaopo Zu, Xueyan Ding, Zhanyong Wei, Hin Chu, Honglei Zhang","doi":"10.1128/mbio.03604-25","DOIUrl":"https://doi.org/10.1128/mbio.03604-25","url":null,"abstract":"<p><p>To establish effective infection, viral pathogens employ diverse strategies through encoded proteins to interfere with host antiviral responses. While previous studies have predominantly focused on elucidating the mechanisms by which individual viral proteins regulate type I interferon (IFN) responses, this study presents the first demonstration that Japanese encephalitis virus (JEV)-encoded NS1 and NS4B proteins cooperatively target the TLR3 receptor signaling pathway to suppress IFN production. Here, we first discovered JEV-encoded multifunctional glycoprotein NS1, which inhibits TLR3-mediated IFN-β production by targeting TLR3 and TRIF. Mechanistically, NS1 interacts with these host factors and may induce their degradation via the autophagy pathway. Building on these findings, we further demonstrated that NS1 and NS4B act synergistically to suppress type I IFN production by enhancing TLR3 and TRIF degradation, thereby facilitating JEV replication. Structural simulation of the NS1-NS4B-TLR3 complex unveiled a dynamic mechanism that NS4B binding induces conformational changes in dimerized NS1, which leads to a tighter binding posture between NS1 and TLR3. Notably, NS4B binding expands the interface between NS1 and TLR3 by 1.5-fold that results in enhanced TLR3 degradation and downstream IFN-β suppression. Functional analysis confirmed that the C291/K293/R314 triple mutation in NS1 synergistically impairs TLR3 degradation. Collectively, using JEV as a model, this study reveals a novel mechanism by which two viral components can act synergistically to evade the host antiviral response. Given the common coexistence and functional interplay of viral-encoded proteins in naturally infected cells, this study establishes a framework for investigating cooperative interactions among multiple viral proteins.</p><p><strong>Importance: </strong>Viruses evade host immunity through encoded viral proteins, while previous research has predominantly focused on single protein mechanisms. This study reveals a novel cooperative immune evasion strategy, demonstrating for the first time that Japanese encephalitis virus NS1 and NS4B proteins act synergistically to degrade the host's TLR3 and TRIF adaptor, thereby suppressing type I interferon production. Structural simulations show that NS4B induces conformational changes in NS1, enhancing its binding to TLR3 and accelerating its degradation. This work establishes a new paradigm for how multiple viral components can function cooperatively to subvert antiviral defenses. Given that viral proteins naturally coexist and interact, this study provides a crucial framework for investigating complex viral protein interplay, which is fundamental to understanding viral pathogenesis.</p>","PeriodicalId":18315,"journal":{"name":"mBio","volume":" ","pages":"e0360425"},"PeriodicalIF":4.7,"publicationDate":"2026-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}