PLoS Pathogens最新文献

{"title":"UBXN7 facilitates SARS-CoV-2 replication via inhibiting the K48-linked ubiquitination of viral N protein.","authors":"Tian Xia, Min Luo, Yuncheng Wang, Yaping Qin, Xiaoning Li, Shuying Chen, Junqi Xiang, Shanrong Yang, Yaokai Wang, Jing Zhu, Bo Yang, Li Lin, Jiajun Yan, Yunxiao Dou, Jian Shang, Na Zang, Yong Lin, Xiaohong Yao, Yushun Wan","doi":"10.1371/journal.ppat.1013593","DOIUrl":"10.1371/journal.ppat.1013593","url":null,"abstract":"<p><p>Host factor-mediated post-translational modification of coronavirus proteins has been demonstrated as an important strategy for regulating viral proliferation. Identification of key host genes involved in this process may provide potential therapeutic targets. In this study, we used the complementary reverse genetic system to determine that UBXN7 promotes SARS-CoV-2 viral double-stranded RNA (dsRNA) production and also promotes the replication of other human coronaviruses. However, UBXN7 does not affect the replication of VSV and RSV, suggesting that it may be a potential pan human coronaviral anti-infection target. Our results revealed that UBXN7 did not affect the viral invasion of cells, but instead hijacked viral genome assembly by interacting with SARS-CoV-2 N protein via its UBX domain. Further data indicated that UBXN7 inhibits K48-linked ubiquitination and proteasomal degradation of SARS-CoV-2 N protein, leading to N protein accumulation. Moreover, K257 of N protein was identified as specific target site of UBXN7 which are critical for viral replication. These findings reveal a novel relationship between host gene-mediated protein ubiquitylation and viral genome assembly, providing new strategies for potential pan-coronavirus drug design.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013593"},"PeriodicalIF":4.9,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12520364/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Myosin-9 is required for lysosome-mediated nonlytic reovirus egress.","authors":"Isabel Fernández de Castro, Martin Sachse, Gwen M Taylor, José J Fernández, Raquel Tenorio, Sara Y Fernández-Sánchez, Terence S Dermody, Cristina Risco","doi":"10.1371/journal.ppat.1013597","DOIUrl":"10.1371/journal.ppat.1013597","url":null,"abstract":"<p><p>Mammalian orthoreoviruses (reoviruses) are nonenveloped, double-stranded RNA viruses that assemble progeny particles in cytoplasmic viral factories (VFs) and exit some types of cells using a nonlytic release mechanism. In human brain microvascular endothelial cells (HBMECs), progeny reovirus virions are selectively sorted from VFs into sorting organelles (SOs), which are derived from lysosomes. Smaller membranous carriers (MCs) bud from SOs and transport progeny virions to the plasma membrane where they are released nonlytically by fusion of MCs with the plasma membrane. To discover cellular factors required for lysosomal modification and nonlytic egress, we used mass spectrometry to identify proteins associated with lysosomes purified from uninfected and reovirus-infected HBMECs as well as virions purified from HBMECs and L929 cells, which differ in the pathways used by reovirus for egress. Network analysis of the proteomic results from HBMECs yielded an enrichment of cytoskeletal proteins centered on myosin-9. Using siRNA gene-silencing of myosin-9, pharmacological inhibition of myosin-9, super-resolution light microscopy, electron microscopy, and three-dimensional electron tomography, we found that myosin-9 acts at late stages of reovirus replication to promote viral egress. Myosin-9 associates with actin filaments attached to mature virions and mediates nonlytic egress of viral progeny from HBMECs. Our findings provide insights into the role of myosin-9 in the intracellular lysosome-mediated reovirus egress pathway and illuminate a new potential therapeutic target for viruses that use this nonlytic egress pathway.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013597"},"PeriodicalIF":4.9,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294140","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phosphorylation of SNAP-25 at Ser187 is enhanced following its cleavage by Botulinum Neurotoxin Serotype A, promoting the dominant-negative effect of the resulting fragment.","authors":"Dilara Koc, Sena Ezgin, Ebru Kavakli, Krishna P Kota, Edanur Sen, Christopher Mahone, Mary Ellen Palko, Lisa H Cazares, Tolga Can, Lino Tessarollo, Sina Bavari, Antoine Marion, Erkan Kiris","doi":"10.1371/journal.ppat.1013604","DOIUrl":"10.1371/journal.ppat.1013604","url":null,"abstract":"<p><p>Botulinum Neurotoxin Serotype A (BoNT/A), responsible for most human botulism cases, inhibits neurotransmitter release by cleaving the target protein SNAP-25. Previous literature demonstrated that BoNT/A mediated cleavage of a small subset of the SNAP-25 pool, resulting in SNAP-25 (1-197) fragments, is sufficient to block exocytosis. SNAP-25 (1-197) potentially competes against intact SNAP-25 for SNARE complexes and blocks neurotransmission through a dominant-negative mechanism. However, how a tiny fraction of cleaved SNAP-25 efficiently outcompetes a large pool of intact SNAP-25 remains unknown. Here, we examined the importance of SNAP-25 phosphorylation at Ser187 residue, located in the C-terminus SNARE domain, in the context of BoNT action. Our results demonstrated that Ser187-phosphorylated SNAP-25 can be efficiently cleaved in cells. Importantly, BoNT/A-cleaved SNAP-25 fragments in neuronal and non-neuronal cells are heavily phosphorylated at Ser187 and localized on the cell membrane. SNAP-25 (1-197) binds to syntaxin-1A, and the interaction is enhanced by Ser187 phosphorylation. We also found that SNAP-25 (1-197) survives longer than the BoNT/A enzymatic component itself in cells. Molecular modeling suggested that SNAP-25 (1-197), phosphorylated or not, forms stable SNARE complexes; however, Ser187 phosphorylation induces local changes in surface electrostatic potential and dynamics of the complex. This study characterizes the molecular mechanism underlying the dominant-negative effect of SNAP-25 (1-197) on neurotransmission. This research could have implications for the future development of BoNT/A inhibitors and the generation of new BoNT/A clinical formulations by regulating the abundance of Ser187 phosphorylation in cleaved SNAP-25 fragments.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013604"},"PeriodicalIF":4.9,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294192","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Is vaccination a feasible public health strategy against fatal Borna disease virus 1 (BoDV-1) encephalitis? An epidemiological perspective.","authors":"Kirsten Pörtner, Christina Frank, Hendrik Wilking, Klaus Stark, Christiane Herden, Martin Beer, Dennis Rubbenstroth, Dennis Tappe","doi":"10.1371/journal.ppat.1013571","DOIUrl":"10.1371/journal.ppat.1013571","url":null,"abstract":"<p><p>Human Borna disease virus 1 (BoDV-1) encephalitis is characterized by rapid clinical progression, an absence of a causal therapy and an extremely high case fatality rate. Here, we discuss prevention options through a hypothetical vaccine focusing on epidemiological features.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013571"},"PeriodicalIF":4.9,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12517474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145287337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mucosal Taï Forest virus infection causes disease in ferrets.","authors":"Paige Fletcher, Kyle L O'Donnell, Joseph F Rhoderick, Corey W Henderson, Atsushi Okumura, Trenton Bushmaker, Kathleen Cordova, Greg Saturday, Andrea Marzi","doi":"10.1371/journal.ppat.1013579","DOIUrl":"10.1371/journal.ppat.1013579","url":null,"abstract":"<p><p>The filovirus Taï Forest virus (TAFV) caused a single human case of infection originating from a chimpanzee outbreak, demonstrating that humans are susceptible to TAFV infection. Existing animal disease models use intramuscular (IM) infection; however, natural filovirus infection likely occurs mucosal. We aimed to develop a ferret disease model by inoculation of TAFV by the IM, intranasal (IN), or aerosol routes. The IM group showed minimal signs of disease while IN and aerosol inoculations resulted in moderate to severe disease and partial lethality. The surviving IN or IM TAFV-infected ferrets were rechallenged IM or IN with Ebola virus (EBOV) as a pilot study assessing the cross-protection potential between these closely related viruses. Only ferrets IN-inoculated with TAFV and IN-inoculated with EBOV were protected from disease, all others succumbed to disease after EBOV infection. This data shows that ferrets are a feasible model to assess TAFV pathogenicity by mucosal exposure routes and that possible cross-protection between TAFV and EBOV may be achieved upon mucosal exposure.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013579"},"PeriodicalIF":4.9,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12530580/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145287327","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly accurate protein structure prediction-based virtual docking pipeline accelerating the identification of anti-schistosomal compounds.","authors":"Wenjun Cheng, Mengjie Gu, Yuepeng Wang, Jing Wang, Shan Li, Gongwen Chen, Xu Chen, Oyetunde T Oyeyemi, Yang Hong, Wei Hu, Jipeng Wang","doi":"10.1371/journal.ppat.1013274","DOIUrl":"10.1371/journal.ppat.1013274","url":null,"abstract":"<p><p>Schistosomiasis is a major neglected tropical disease that lacks an effective vaccine and faces increasing challenges from praziquantel resistance, underscoring the urgent need for novel therapeutics. Target-based drug discovery (TBDD) is a powerful strategy for drug development. In this study, we utilized AlphaFold to predict the structures of target proteins from Schistosoma mansoni and S. japonicum, followed by virtual molecular screening to identify potential inhibitors. Among 202 potential therapeutic targets, we identified 37 proteins with high-accuracy structural predictions suitable for molecular docking with 14,600 compounds. This screening yielded 268 candidate compounds, which were further evaluated ex vivo for activity against both adult and juvenile S. mansoni and S. japonicum. Seven compounds exhibited strong anti-schistosomal activity, with HY-B2171A (Carubicin hydrochloride, CH) emerging as the most potent. CH was predicted to target the splicing factor U2AF65, and knockdown of its coding gene Smp_019690 resulted in a phenotype similar to CH treatment. RNA sequencing revealed that both CH treatment and Smp_019690 RNA interference (RNAi) disrupted splicing events in the parasites. Further studies demonstrated that CH impairs parasite viability by inhibiting U2AF65 function in mRNA splicing regulation. By integrating RNAi-based target identification with structure-based virtual screening, alongside ex vivo phenotypic and molecular analyses of compound-treated schistosomes, our study provides a comprehensive framework for anti-schistosomal drug discovery and identifies promising candidates for further preclinical development.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013274"},"PeriodicalIF":4.9,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12533970/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145287361","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HTLV-1 p12 is cleaved to p8 by the signal peptidase complex and its inhibition impairs p8-dependent transmission.","authors":"Florian Simon, Norbert Donhauser, Laura M Kemeter, Franziska Wittdorf, Sebastian Millen, Heinrich Sticht, Andrea K Thoma-Kress","doi":"10.1371/journal.ppat.1013570","DOIUrl":"10.1371/journal.ppat.1013570","url":null,"abstract":"<p><p>Infection with the oncogenic delta-retrovirus Human T-cell Leukemia Virus Type 1 (HTLV-1) causes aggressive CD4 + T-cell malignancy or progressive neuroinflammatory disorders after a long latency period. The HTLV-1 accessory protein p8 is cleaved from its precursor p12, and both proteins are required for viral persistence. Moreover, p8 enhances viral infectivity by inducing cell-cell contacts and cellular conduit formation. To date, host factors cleaving p12 to p8 remain unknown. Here, we report that p12 carries a signal peptide that is cleaved by the signal peptidase complex (SPC) to generate p8, and blocking of p12 cleavage correlated with a decreased cell aggregation and conduit formation, leading to impaired viral transmission of chronically HTLV-1 infected MT-2 cells. Bioinformatics identified p12 to carry a signal peptide, which is cleaved to generate p8. Inhibition of the SPC function by the SPC-specific inhibitor Cavinafungin and transient knockdown of SPC subunits confirmed the importance of the SPC to cleave p12 to p8. Mutational studies of the signal peptide sequence based on bioinformatics predictions generated cleavage-deficient p12 mutants and verified critical residues for signal peptide cleavage. Further, co-culture assays between Cavinafungin pre-treated chronically infected MT-2 cells, which transmit HTLV-1 dependent on p8, and Jurkat T-cells revealed a significantly impaired viral cell-cell transmission, suggesting that blockage of p12 cleavage interferes with p8-dependent HTLV-1 transmission. Imaging analysis confirmed that SPC-inhibition impairs cell aggregation in MT-2 cells and blocks p8-induced conduit formation in transfected Jurkat T-cells. Collectively, we identified the SPC as the host cell factor cleaving p12 to p8. Inhibition of p12 cleavage led to the absence of p8, which led to impaired cell-to-cell transmission, and coincided with the absence of p8-induced cell aggregation and conduit formation.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013570"},"PeriodicalIF":4.9,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12513584/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145276352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lactic acid produced by optimal vaginal Lactobacillus spp. potently and specifically inactivates HIV-1 in vitro by targeting the viral RNA genome and reverse transcriptase.","authors":"Muriel Aldunate, David Tyssen, Adam Johnson, Catherine F Latham, Paula Ellenberg, Nathan Cowieson, Joshua A Hayward, Rob J Center, Paul A Ramsland, Anna C Hearps, Gilda Tachedjian","doi":"10.1371/journal.ppat.1013594","DOIUrl":"10.1371/journal.ppat.1013594","url":null,"abstract":"<p><p>Vaginal microbiota modulates susceptibility to sexually transmitted infections and produces carboxylic acid metabolites that have antimicrobial activity; however, their activity against viral sexually transmitted infections is not well defined. We determined the HIV-1 virucidal activity of lactic acid (LA), short chain fatty acids (SCFAs), and succinic acid, representing conditions observed in women with an optimal Lactobacillus-dominated vaginal microbiota compared to women with bacterial vaginosis. Virucidal activity against enveloped HIV-1 and HSV-2, the non-enveloped HPV16, and the mechanism by which LA inactivates HIV-1 was further assessed. LA was > 10-fold more potent at inactivating an HIV-1 transmitted/founder strain than SCFAs and succinic acid when tested at an equivalent 20  mM of protonated acid (p≤0.05). While LA decreased HIV-1 infectivity by >103-fold, virions were intact, expressed a similar gp120:p24 ratio, and showed only a 2-fold decrease in CD4 binding compared to untreated HIV-1 (p≤0.05). Treatment of recombinant gp120 with LA revealed no major conformational changes by small angle X-ray scattering. LA treatment of HIV-1 resulted in an 80% decrease in virion-associated reverse transcriptase activity compared to untreated virus (p < 0.01), which was more potent than acetic acid or HCl-adjusted media at the same pH, with this effect observed in the presence of cervicovaginal fluid. LA decreased HIV-1 virion-associated RNA levels by ∼50% compared to untreated virus (p < 0.001), acetic acid or HCl acidified media. In contrast, HSV-2 virucidal activity of LA was similar to acetic acid and HCl-acidified media while HPV16 was acid-resistant. Our results demonstrate LA's potent and specific HIV-1 virucidal activity compared to SCFAs and succinic acid found in the female reproductive tract, and its HIV-1 virucidal mechanism mediated by penetration of the viral membrane and core to target a key viral enzyme and nucleic acid. These findings have implications for the vaginal transmission of HIV to partners and neonates during birth.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013594"},"PeriodicalIF":4.9,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12527216/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145276345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TfR1 facilitates influenza virus endocytosis and uncoating by interacting with NA and M1 via extracellular and intracellular domains.","authors":"Xinchen Wang, Yuanhao Li, Dezhong Ji, Yiming Wang, Xiaoyang Wang, Kangming Guo, Mengyang Wang, Yu Mu, Chen Qin, Tao Yuan, Yuanjie Zhang, Zhiqian Chen, Xingxing Zhu, Xiaohui Zhang, Honghui Jiang, Qiuchen He, Chuanling Zhang, Sulong Xiao, Lihe Zhang, Demin Zhou","doi":"10.1371/journal.ppat.1013511","DOIUrl":"10.1371/journal.ppat.1013511","url":null,"abstract":"<p><p>An intriguing enigma in virology is the utilization of transferrin receptor 1 (TfR1) by various viruses as an entry portal into host cells, a mechanism that remains relatively underexplored. In this study, we report a strategy to investigate the multifaceted aspects of viral entry, using Influenza A viruses (IAVs) as a model system. By decorating the sialylated viral envelope with photo-crosslinking moieties, we identify and elucidate the pivotal role of TfR1 in this process. Our results demonstrate that TfR1 initially functions as a receptor, interacting with the viral neuraminidase (NA) through its extracellular apical domain, thereby initiating viral endocytosis. Subsequently, TfR1 acts as a matrix degradator, engaging its intracellular stop-transfer sequence with the viral matrix protein 1 (M1), which in turn triggers proteasome- and aggresome-mediated nucleocapsid uncoating. The identification of the molecular interactions between TfR1 and NA, as well as the reciprocal degradation of TfR1 and M1 not only illuminates a cellular pathway that enriches our understanding of viral entry mechanisms but also presents exciting avenues for the development of innovative antiviral strategies beyond IAVs.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013511"},"PeriodicalIF":4.9,"publicationDate":"2025-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12533913/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145276364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The gonococcal vaccine candidate antigen NGO1701 is a N. gonorrhoeae periplasmic copper storage protein.","authors":"Shea K Roe, Rafał Mazgaj, Tianmou Zhu, Mariam Esmaeeli, Lisa A Lewis, Caroline Genco, Kevin J Waldron, Paola Massari","doi":"10.1371/journal.ppat.1013559","DOIUrl":"10.1371/journal.ppat.1013559","url":null,"abstract":"<p><p>The increasing worldwide trend of antibiotic-resistant Neisseria gonorrhoeae strains highlights the urgent need for new therapeutic strategies against this sexually transmitted pathogen, including a gonococcal vaccine. We previously designed a bioinformatics-based candidate selection pipeline (CASS) and identified potential novel gonococcal vaccine targets among hypothetical proteins expressed during natural human infection. One of these candidates, NGO1701, is a predicted periplasmic four-helix bundle protein with amino acid sequence homology to the copper storage protein 1 (Csp1) from Methylosinus trichosporium OB3b. In this study, we confirmed that purified NGO1701 binds 15 Cu(I) ions per monomer in vitro, supporting its function as Csp in N. gonorrhoeae. Using a ngo1701 deletion mutant generated in N. gonorrhoeae F62, we investigated its role in bacteria physiology. We showed that ablation of Csp was not limiting for bacterial growth and fitness in vitro, but the Δcsp strain became significantly more susceptible to copper mediated toxicity. This phenotype was rescued by csp gene complementation, indicating a role in protection against copper toxicity. Our results indicate that Csp participates in periplasmic copper homeostasis in N. gonorrhoeae, buffering excess copper to reduce toxicity and playing a putative role in copper delivery to important copper-enzymes. Csp does not appear to be involved in bacterial host cell interaction and activation in vitro, since no difference in the ability of N. gonorrhoeae to adhere/invade epithelial cells or induce IL-8 secretion was reported among wild type, csp deletion mutant and complemented strains. Furthermore, sera from mice immunized with NGO1701 failed to recognize Δcsp by dot blot and ELISA, and the sera's ability to kill N. gonorrhoeae was abrogated against Δcsp. However, both functions were restored after gene complementation, supporting the relevance of Csp as a potential vaccine target. Allelic analysis of Neisseria species revealed that this gene is absent in N. meningitidis, thus making it a gonococcal-specific target.</p>","PeriodicalId":48999,"journal":{"name":"PLoS Pathogens","volume":"21 10","pages":"e1013559"},"PeriodicalIF":4.9,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12510493/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145259421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}