Journal of Virology最新文献

{"title":"Advances in African swine fever virus molecular biology and host interactions contributing to new tools for control.","authors":"Linda K Dixon","doi":"10.1128/jvi.00932-24","DOIUrl":"10.1128/jvi.00932-24","url":null,"abstract":"<p><p>African swine fever virus (ASFV) causes a frequently fatal hemorrhagic disease in domestic pigs and wild boar. The spread from Africa to Georgia in 2007 initiated a pandemic affecting many European and most Asian countries. This has had a very high socio-economic impact and threatens global food security. The virus is a large, complex, cytoplasmic DNA virus, the only member of the <i>Asfarviridae</i> family and codes for 170-190 proteins. Many of these have unknown functions and do not resemble other viruses or host proteins. This complexity has hindered the development of vaccines and other tools for control. The intensity of research has increased since the spread of ASFV in Europe and Asia, leading to rapid advances in knowledge. This review summarizes recent research, including the determination by cryogenic electron microscopy of the virus capsid structure and virion proteome. Novel information on the virus replication cycle, including mechanisms of virus entry into cells and the identification of host endosomal proteins important for entry, is summarized. Multiple, novel virus immune evasion proteins and their targets in the type I interferon response and inflammation pathways have been identified. The potential for the application of this knowledge to developing novel control tools, including modified live vaccines and other interventions targeting critical virus processes or host interactions, is discussed.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0093224"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172490/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143975413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The NSP6-L260F substitution in SARS-CoV-2 BQ.1.1 and XBB.1.16 lineages compensates for the reduced viral polymerase activity caused by mutations in NSP13 and NSP14.","authors":"Yuri Furusawa, Kiyoko Iwatsuki-Horimoto, Seiya Yamayoshi, Yoshihiro Kawaoka","doi":"10.1128/jvi.00656-25","DOIUrl":"10.1128/jvi.00656-25","url":null,"abstract":"<p><p>Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variants emerged at the end of 2021, and their subvariants are still circulating worldwide. While changes in the S protein of these variants have been extensively studied, the roles of amino acid substitutions in non-structural proteins have not been fully revealed. In this study, we found that SARS-CoV-2 bearing the NSP6-L260F substitution emerged repeatedly when we generated several SARS-CoV-2 variants by reverse genetics or when we passaged SARS-CoV-2 isolated from clinical samples and that it was selected under cell culture conditions. Although this substitution has been detected in BQ.1.1 and XBB.1.16 that circulated in nature, its effect on viral properties is unclear. Here, we generated SARS-CoV-2 with or without the NSP6-L260F by reverse genetics and found that NSP6-L260F promotes virus replication <i>in vitro</i> and <i>in vivo</i> by increasing viral polymerase activity and enhancing virus pathogenicity in hamsters. We also identified disadvantageous substitutions, NSP13-M233I and NSP14-D222Y, that reduced BQ.1.1 and XBB.1.16 replication, respectively. These adverse effects were compensated for by NSP6-L260F. Our findings suggest the importance of NSP6-L260F for virus replication and pathogenicity and reveal part of the evolutionary process of Omicron variants.IMPORTANCEAlthough the properties of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variants continue to change through the acquisition of various amino acid substitutions, the roles of the amino acid substitutions in the non-structural proteins have not been fully explored. In this study, we found that the NSP6-L260F substitution enhances viral polymerase activity and is important for viral replication and pathogenicity. In addition, we found that the NSP13-M233I substitution in the BQ.1.1 lineage and the NSP14-D222Y substitution in the XBB.1.16 lineage reduce viral polymerase activity, and this adverse effect is compensated for by the NSP6-L260F substitution. Our results provide insight into the evolutionary process of SARS-CoV-2.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0065625"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172475/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143975416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Classical swine fever virus utilizes stearoyl-CoA desaturase 1-mediated lipid metabolism to facilitate viral replication.","authors":"Ji-Shan Bai, Lin-Ke Zou, Ya-Yun Liu, Lin-Han Zhong, Jing Chen, Jin-Xia Chen, Bing-Qian Zhao, Rong-Chao Liu, Bo-Tao Sun, Bin Zhou","doi":"10.1128/jvi.00551-25","DOIUrl":"10.1128/jvi.00551-25","url":null,"abstract":"<p><p>Viral infections can significantly alter cellular lipid metabolism by modulating key rate-limiting enzymes, including fatty acid synthase (FASN), stearoyl-CoA desaturase 1 (SCD1), and acetyl-CoA carboxylase (ACC). Our previous study revealed the pivotal role of FASN in lipid droplet (LD) synthesis and the promotion of classical swine fever virus (CSFV) replication. However, the roles of the other two key enzymes in CSFV infection remain unexplored. In this study, we screened a library of 96 lipid metabolism-targeted compounds and identified an antiviral inhibitor of SCD1, a rate-limiting enzyme in monounsaturated fatty acid synthesis, that inhibits CSFV replication. Suppressing SCD1 activity through inhibitors or small interfering RNA knockdown reduces CSFV proliferation. However, this suppression is reversed by adding SCD1 active products (oleic acid/palmitoleic acid [OA/PA]), highlighting the essential role of SCD1 in CSFV proliferation. Mechanistically, CSFV non-structural protein p7 interacts with SCD1 and recruits it to the viral replication complex (VRC) during infection. Importantly, CSFV infection activates the endoplasmic reticulum stress pathway IRE1α/XBP1, which positively regulates SCD1 expression, leading to increased production of triglyceride (TG) and LDs and subsequently enhancing CSFV replication. In summary, our study elucidates the critical role of SCD1 in the CSFV life cycle and highlights its potential as an antiviral target for developing new therapies against <i>Flaviviridae</i>.IMPORTANCEUnderstanding the virus's pathogenesis within the host is essential for advancing antiviral therapeutics and vaccine development. Previous studies have demonstrated that classical swine fever virus (CSFV) leverages host lipid metabolic rate-limiting enzymes, such as fatty acid synthase (FASN), to support viral replication. This study identified stearoyl-CoA desaturase 1 (SCD1), a key enzyme in monounsaturated fatty acid biosynthesis, as a novel regulator of CSFV replication. Mechanistically, the viral non-structural protein p7 mediates the recruitment of SCD1 to the endoplasmic reticulum (ER), facilitating the formation of viral replication complexes (VRCs). Additionally, our findings showed that viral infection activated the ER stress pathway IRE1α/XBP1, which upregulated SCD1 expression and promoted the synthesis of triglycerides (TG) and lipid droplets (LDs). This study provides insights into the metabolic reprogramming triggered by viral infection to support replication and underscores the intricate crosstalk between ER stress and lipid metabolism during CSFV infection. These findings have significant implications for identifying novel antiviral targets against CSFV.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0055125"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172442/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144094075","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<u>Ad</u>vanced <u>v</u>iral genome <i><u>i</u>n vitro</i> <u>C</u>as9 <u>e</u>diting (AdVICE): an overnight method for traceless and limitless manipulation of adenoviral and vector genomes with large transgenes.","authors":"Jean-Baptiste Vergnes, Benoit Roger, Richard Iggo, Harald Wodrich","doi":"10.1128/jvi.02265-24","DOIUrl":"10.1128/jvi.02265-24","url":null,"abstract":"<p><p>The size and complexity of large viral genomes limit the technical possibilities for genome manipulations in fundamental research and medical or technological applications. State-of-the-art recombineering in bacteria has partially overcome this limit but remains a time-consuming and complex procedure requiring specialist expertise. Here, we describe a simplified and highly efficient <i>in vitro</i> protocol for unlimited and traceless manipulation applicable to large viral genomes from DNA viruses using a combination of CRISPR/Cas9 cleavage and <i>in vitro</i> DNA assembly. We successfully used the protocol to manipulate adenovirus genomes, showing that genome rescue from viruses, insertions, deletions, and mutagenesis can be performed in a simple overnight procedure in a standard laboratory setting without the need for advanced knowledge of molecular biology. Finally, we use our approach to demonstrate the <i>de novo</i>, multi-step construction of an adenovirus vector suitable for delivering very large transgenes for gene editing.IMPORTANCEThe 36 kb size of the adenoviral genome has long been a deterrent to the construction of adenoviral mutants by scientists wishing to study the virus itself or to construct adenoviral vectors for cell biology and gene therapy. Most previous techniques, such as recombineering and yeast gap repair, impress more by their elegance than by their ease. In this paper, we use Cas9 ribonucleoprotein particles (RNPs) to target cleavage to specific sites in an adenoviral plasmid, then repair the break by Gibson assembly. Gibson assembly with synthetic DNA fragments has transformed basic cloning. Combining it with Cas9 RNPs, which act like highly specific restriction enzymes, makes adenoviral mutagenesis as easy as traditional plasmid cloning. We have used the approach to modify multiple sites in the adenoviral genome, but it could be applied to any large DNA virus for which the genome can be cloned in a plasmid.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0226524"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172474/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144111128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of SARS-CoV-2 intrahost genetic evolution in vaccinated and non-vaccinated patients from the Kenyan population.","authors":"Doreen Lugano, Kennedy Mwangi, Bernard Mware, Gilbert Kibet, Shebbar Osiany, Edward Kiritu, Paul Dobi, Collins Muli, Regina Njeru, Tulio de Oliveira, M Kariuki Njenga, Andrew Routh, Samuel O Oyola","doi":"10.1128/jvi.00482-25","DOIUrl":"10.1128/jvi.00482-25","url":null,"abstract":"<p><p>Vaccination is a key control measure of coronavirus disease 2019 by preventing severe effects of disease outcomes, reducing hospitalization rates and death, and increasing immunity. However, vaccination can affect the evolution and adaptation of SARS-CoV-2 largely through vaccine-induced immune pressure. Here, we investigated intrahost recombination and single nucleotide variations (iSNVs) on the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome in non-vaccinated and vaccinated sequences from the Kenyan population to profile intrahost viral genetic evolution and adaptations driven by vaccine-induced immune pressure. We identified recombination hotspots in the S, N, and ORF1a/b genes and showed the genetic evolution landscape of SARS-CoV-2 by comparing within- and inter-wave recombination events from the beginning of the pandemic (June 2020 to December 2022) in Kenya. We further reveal differential expression of recombinant RNA species between vaccinated and non-vaccinated individuals and perform an in-depth analysis of iSNVs to identify and characterize the functional properties of non-synonymous mutations found in ORF-1 a/b, S, and N genes. Lastly, we detected a minority variant in non-vaccinated patients in Kenya, with an immune escape mutation S255F of the spike gene, and showed differential recombinant RNA species. Overall, this work identified unique <i>in vivo</i> mutations and intrahost recombination patterns in SARS-CoV-2, which could have significant implications for virus evolution, virulence, and immune escape.IMPORTANCEThe impact of vaccination on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genetic diversity in Kenya and much of Africa remains unknown. This can be attributed to lower sequencing rates; however, this information is relevant to improvement in vaccine and antiviral research. In this study, we investigated how vaccination and SARS-CoV-2 transmission waves affect intrahost non-homologous recombination and single nucleotide variations (iSNVs). We identified unique <i>in vivo</i> mutations and intrahost recombination patterns in SARS-CoV-2, which could have significant implications for virus evolution, virulence, and immune escape. We also demonstrate a methodology for studying genetic changes in a pathogen by a simultaneous analysis of both intrahost single nucleotide variations and recombination events. The study reveals the diversity of SARS-CoV-2 in Kenya and highlights the need for sustained genomic surveillance in Kenya and Africa to better understand how the virus evolves. Such surveillance ensures detection of drifts in evolution, allowing information for updates in vaccines, policy making, and containment of future variants of SARS-CoV-2.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0048225"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172480/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144024410","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dissection of the global responses of mandarin fish pyloric cecum to an acute ranavirus (MRV) infection reveals the formation of serositis and then ascites.","authors":"Wenfeng Zhang, Yong Li, Xiaosi Wu, Qianqian Sun, Yuting Fu, Shaoping Weng, Jianguo He, Chuanfu Dong","doi":"10.1128/jvi.02308-24","DOIUrl":"10.1128/jvi.02308-24","url":null,"abstract":"<p><p>Mandarin fish ranavirus (MRV), a new member of the species <i>Ranavirus micropterus1</i>, sharing over 98% whole-genome nucleotide identity with the well-known largemouth bass virus (LMBV), is a distinct member of the genus <i>Ranavirus</i> within the family <i>Iridoviridae</i>. Our recent work showed that acute MRV infection predominantly affects the pyloric cecum, a critical visceral organ in mandarin fish, and was hypothesized to drive the characteristic external clinical sign of severe ascites. In this study, we reveal that acute MRV infection initially targets the serosal layer of the pyloric cecum of mandarin fish, leading to rapid progression into fibrinous serositis characterized by serosal hypertrophy, fibrosis, hyperemia, edema, and tissue adhesions. Using single-cell RNA sequencing, we dissect the cellular composition of epithelial, immune, and stromal populations, identifying significant enrichment of macrophages and granulocytes, alongside T and natural killer cells, as key mediators of acute cytokine and inflammatory responses. Then, robust experimental evidence demonstrates that MRV infects specific immune cell subsets of T and B cells and stromal cells of fibroblasts, myofibroblasts, endothelial cells, and pericytes, resulting in upregulation of genes and pathways associated with extracellular matrix (ECM) formation, collagen biosynthesis, and vascular remodeling in the hyperplastic serosal zone. Additionally, both host-derived type V collagens and MRV-encoded collagens are implicated in ECM formation in the hypertrophic serosa. Collectively, this study provides a comprehensive single-cell resolution analysis of the pyloric cecum's response to acute MRV infection and highlights virus-driven serositis as the underlying cause of severe ascites in mandarin fish.IMPORTANCEThe pyloric cecum is a vital digestive and immune organ in many bony fish species, including the mandarin fish, a carnivorous species with an exceptionally developed pyloric cecum comprising 207-326 ceca per individual. While MRV/LMBV infects various fish species, severe ascites is uniquely observed in infected mandarin fish. This study demonstrates that acute MRV infection induces fibrinous serositis in the pyloric cecum, characterized by hyperemia, edema, and hyperplasia, ultimately resulting in ascites and mortality. Leveraging single-cell RNA sequencing, we provide a detailed landscape of the cell types affected or involved in the inflammatory response, revealing their roles in the pathogenesis of serositis. These findings advance our understanding of MRV-induced pathology and its species-specific manifestations.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0230824"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172472/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143978020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rescue of naïve porcine circovirus type 3 and its pathogenesis in CD pigs.","authors":"Baoge Zhang, Jinshuang Cai, Chenguang Zhu, Yuyu Zhang, Jiaqiang Wu, Yufeng Li","doi":"10.1128/jvi.00341-25","DOIUrl":"10.1128/jvi.00341-25","url":null,"abstract":"<p><p>The pathogenesis of naïve porcine circovirus type 3 (PCV3) in pigs is not accurately elucidated due to virus isolation failure <i>in vitro</i>. In this study, naïve PCV3 was successfully rescued and used to infect cesarean-derived (CD) pigs. The PCV3 genome was synthesized, cloned, and inserted into the pBluescript SK vector. PK-15 cells transfected with the recombinant plasmid pSK-PCV3 were passaged, and Cap protein expression was confirmed by immunofluorescence assay (IFA) and western blotting. Virus particles (~20 nm) were observed via transmission electron microscopy (TEM). The viral growth curve was plotted to determine the replication of the virus within the cells. Nocodazole treatment demonstrated that PCV3 replication is dependent on microtubule polymerization in the cell. Cells infected with PCV3 or PCV3-positive clinical samples (wPCV3) showed only cytoplasmic fluorescence. PCV3 can successfully infect CD pigs, resulting in persistent viremia, and increased viral loads in tissues and an antibody delay were observed. Inflammatory lesions were also observed in the lungs, lymph nodes, livers, and intestines of infected pigs. Stimulation of peripheral blood mononuclear cells (PBMCs) with virus-like particles (VLPs) containing the Cap protein significantly inhibited cell proliferation. scRNA-seq revealed a significant reduction in T helper 2 (Th2) cells and the migration of T helper 1 (Th1) cells toward the late differentiation stage following infection. In particular, the decrease in Th2 cells may indicate impaired humoral immunity, leading to delayed antibody production and immunosuppression. Our study is the first to observe PCV3 via TEM and to elucidate its immunosuppressive mechanisms in CD pigs.</p><p><strong>Importance: </strong>This study is of great significance as it successfully rescued naïve PCV3 and, for the first time, observed clear PCV3 viral particles using transmission electron microscopy. The successful infection of CD pigs deepened our understanding of its pathogenic mechanisms. The results revealed key aspects of viral replication, the impact of the virus on immune responses, and associated inflammatory lesions in various tissues. Notably, the study found that the reduction of Th2 cells leads to impaired humoral immunity and delayed antibody production, which may provide valuable insights for vaccine development and swine disease management.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0034125"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172495/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143989421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"IFITM proteins are key entry factors for porcine epidemic diarrhea coronavirus.","authors":"Lilei Lv, Huaye Luo, Jingxuan Yi, Kang Zhang, Yanhua Li, Wu Tong, Yifeng Jiang, Yanjun Zhou, Guangzhi Tong, Changlong Liu","doi":"10.1128/jvi.02028-24","DOIUrl":"10.1128/jvi.02028-24","url":null,"abstract":"<p><p>Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that poses a substantial threat to the global swine industry. However, our current understanding of the host factors crucial for PEDV infection remains limited. To identify these host factors, we conducted a genome-wide CRISPR/Cas9 gene knockout screen using a PEDV-permissive cell line. Our results indicate that the endogenous expression of human interferon-inducible transmembrane protein 3 (IFITM3) enhances PEDV entry and replication. Silencing or eliminating endogenous IFITM3 in Huh7 cells significantly suppressed PEDV entry, whereas reintroducing IFITM3 partially restored susceptibility to PEDV. Overexpression of human IFITM3 or IFITM2, but not IFITM1, in Huh7.5 cells substantially increased PEDV entry and replication. Importantly, our results suggest that human IFITM3 influences PEDV entry at a later stage. Furthermore, the overexpression of porcine IFITM1 significantly enhanced PEDV infection in LLC-PK1 cells, whereas the overexpression of porcine IFITM2/3 did not produce similar effects. Notably, removing the C-terminal 15 amino acids of porcine IFITM2/3 resulted in increased PEDV entry. Coimmunoprecipitation analyses showed that all IFITMs interacted with the PEDV S1 protein, indicating a direct role in the viral entry process. Additionally, porcine IFITM1 colocalized with the PEDV S protein at the cell nuclear periphery and enhanced PEDV infection in porcine small intestinal organoids. Overall, our results suggest that IFITMs are critical in facilitating PEDV entry into cells. Targeting IFITMs may provide a promising strategy for controlling PEDV transmission and developing interventions to mitigate the virus's impact on the swine industry.</p><p><strong>Importance: </strong>Understanding the mechanisms underlying porcine epidemic diarrhea virus (PEDV) infection is vital for addressing its significant impact on the swine industry. This study reveals that interferon-inducible transmembrane (IFITM) proteins, particularly human IFITM3 and porcine IFITM1, play crucial roles in facilitating PEDV entry and replication. By elucidating these molecular interactions, the research highlights the potential of IFITMs as therapeutic targets for managing PEDV infections and paves the way for antiviral strategies. Moreover, this research extends beyond PEDV management, underscoring the critical role of host factors in controlling the spread of pathogenic coronaviruses.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0202824"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172462/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and characterization of O-GlcNAc modifications of a conserved orthopoxvirus core protein.","authors":"Yunliang Zhang, Bernard Moss","doi":"10.1128/jvi.00058-25","DOIUrl":"10.1128/jvi.00058-25","url":null,"abstract":"<p><p>O-GlcNAcylation, a post-translational modification consisting of O-linked N-acetylglucosamine attached to serine and threonine residues, occurs in thousands of cytoplasmic, nuclear, and mitochondrial proteins but has been reported for relatively few viral proteins. We used click chemistry, specific antibodies, and mass spectrometry to investigate the O-GlcNAcylation of vaccinia virus (VACV) proteins. A virion protein of ~40 kDa was identified by SDS-polyacrylamide gel electrophoresis following azide-alkyne cycloaddition of biotin or an infrared dye to O-GlcNAc residues. Candidate O-GlcNAc virion proteins were detected by mass spectrometry, and A4, a highly conserved core component required for virion assembly, was identified by decreased electrophoretic mobility resulting from the specific attachment of multiple 10 kDa polyethylene glycol residues to O-GlcNAc sites. O-GlcNAc was not detected in virions of an A4 deletion mutant, suggesting A4 is the only or major constituent with this modification. Multiple O-GlcNAc modified amino acids in intrinsically disordered regions of A4 were identified by electron transfer dissociation mass spectrometry. Recombinant A4 was O-GlcNAcylated following stable and transient transfection of uninfected cell lines, suggesting a role for a cellular enzyme, which was confirmed by reduction of the modification by specific inhibitors of O-GlcNAc transferase during virus infection. Moreover, induced degradation of O-GlcNAc transferase prior to VACV infection decreased O-GlcNAcylation of A4 to undetectable levels without diminishing the A4 abundance. Nevertheless, the specific infectivity of O-GlcNAc-deficient virus particles was unimpaired. O-GlcNAcylation either has a subtle role in the VACV life cycle, or A4 is an inadvertent substrate of the promiscuous O-GlcNAc transferase.IMPORTANCEO-GlcNAc is a reversible enzymatic post-translational modification of serine and threonine residues found on thousands of cellular proteins with roles in regulating numerous functions including signal transduction, transcription, and stress response. However, little is known about O-GlcNAc modifications of viral proteins. Here, we report that the vaccinia virus A4 core protein has multiple O-GlcNAc modifications. The cellular O-GlcNAc transferase was shown to be required for modifying the vaccinia virus protein, which is synthesized and assembled into virus particles within cytoplasmic virus factories. Moreover, inhibition and degradation of the transferase prevented O-GlcNAcylation of A4. Nevertheless, virus assembly and replication <i>in vitro</i> were unaffected by the absence of the modification, suggesting that the addition of O-GlcNAc to A4 has a subtle role or that the modification is a byproduct of a promiscuous O-GlcNAc transferase that preferentially modifies intrinsically disordered regions of proteins.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0005825"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172419/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144128027","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Palmitoylation enhances the stability of porcine epidemic diarrhea virus spike protein by antagonizing its degradation via chaperone-mediated autophagy to facilitate viral proliferation.","authors":"Qisheng Qian, Shuang-Shuang Zhao, Lei Yang, Guangxu Xing, Yumei Chen, Chao Liang, Haili Wang, Rui Li, Songlin Qiao, Aiping Wang, Gaiping Zhang","doi":"10.1128/jvi.00347-25","DOIUrl":"10.1128/jvi.00347-25","url":null,"abstract":"<p><p>Porcine epidemic diarrhea (PED) is a highly pathogenic and infectious intestinal disease caused by the PED virus (PEDV) and has inflicted substantial economic losses on the global swine industry. Therefore, it is imperative to explore appropriate targets to restrain PEDV infection. PEDV spike (S) protein is crucial for viral infection and is regarded as an ideal target for the development of vaccines and antiviral therapeutics. Palmitoylation is a significant post-translational modification implicated in multiple viral replication cycles. Despite the fact that palmitoylation of certain coronavirus S proteins has been reported, the specific biological significance and underlying molecular mechanisms of PEDV S protein palmitoylation have not been fully defined. In the present study, we uncover that palmitoylation enhances the stability of PEDV S protein to promote viral proliferation. Mechanistically, we identify that a cysteine-rich region within the cytoplasmic tail of PEDV S protein is palmitoylated by the zinc finger Asp-His-His-Cys domain palmitoyltransferase 5 (ZDHHC5). We further illustrate that palmitoylation prevents the recognition of Lys-Phe-Glu-Arg-Gln (KFERQ)-like motif in PEDV S protein by heat shock cognate protein of 70 kDa (HSC70), thereby antagonizing its degradation via chaperone-mediated autophagy (CMA). Collectively, our findings underscore the importance of palmitoylation for PEDV pathogenesis and provide prospective targets for the development of antiviral interventions.IMPORTANCEPEDV poses a serious threat to pig farming worldwide. As a consequence, a comprehensive investigation of PEDV pathogenesis is of great significance for the prevention and control of the virus. Here, we verify that ZDHHC5-mediated palmitoylation of PEDV S protein enhances its stability through impeding recognition by HSC70 and antagonizing degradation via CMA to facilitate viral propagation. Our findings highlight the important role of palmitoylation in PEDV proliferation and support palmitoylation as a promising target for the development of antiviral strategies.</p>","PeriodicalId":17583,"journal":{"name":"Journal of Virology","volume":" ","pages":"e0034725"},"PeriodicalIF":4.0,"publicationDate":"2025-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12172468/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144119997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}