The FEBS journal最新文献

{"title":"Potency of agarose gel-supported lipid bilayers for electrophysiologic analysis of channel pores formed by Bacillus thuringiensis insecticidal proteins.","authors":"Tsubasa Okuda, Tomoya Takeuchi, Mami Asakura, Minako Hirano, Toru Ide, Tohru Hayakawa","doi":"10.1111/febs.70070","DOIUrl":"https://doi.org/10.1111/febs.70070","url":null,"abstract":"<p><p>Electrophysiologic analysis using artificial lipid bilayers is useful for studying the formation of pores by insecticidal proteins, especially the ion permeability of toxin pores. However, such studies are time-consuming and require special skills, particularly regarding the construction of lipid bilayers and promoting toxin pore formation. To facilitate the analysis of toxin pore formation in the present study, we evaluated the usefulness of agarose gel-supported lipid bilayers for electrophysiologic measurements using two structurally different mosquito-larvicidal proteins, Mpp46Ab and Cry4Aa. The agarose gel-supported lipid bilayers enabled the measurement of channel currents through pores made by both toxins and, notably, the lipid bilayers could be easily reconstructed even after disruption of the lipid bilayer. Using this system, measurements could be repeated at least five times using the same apparatus and toxins. We also investigated the effect of the lipid bilayer component on toxin pore formation and found that the incorporation of both cholesterol and sphingomyelin into the lipid bilayer facilitates the formation of pores by both Mpp46Ab and Cry4Aa. Both cholesterol and sphingomyelin are major components of lipid raft microdomains, suggesting that, in addition to recruiting toxin receptors, raft microdomains play a key role in membrane insertion and pore formation by insecticidal proteins.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143652819","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A metastasis-associated pannexin-1 mutant (Panx1^1-89) forms a minimalist ATP release channel.","authors":"Junjie Wang, Noah J Levi, Maykelis Diaz-Solares, Carsten Mim, Gerhard Dahl, Rene Barro-Soria","doi":"10.1111/febs.70060","DOIUrl":"10.1111/febs.70060","url":null,"abstract":"<p><p>A truncated form of the ATP release channel pannexin 1 (Panx1), Panx1^1-89, is enriched in metastatic breast cancer cells and has been proposed to mediate metastatic cell survival by increasing ATP release through mechanosensitive Panx1 channels. However, whether Panx1^1-89 on its own [without the presence of wild-type Panx1 (wtPanx1)] mediates ATP release has not been tested. Here, we show that Panx1^1-89 by itself can form a constitutively active membrane channel, capable of releasing ATP even in the absence of wtPanx1. Our biophysical characterization reveals that most basic structure-function features of the channel pore are conserved in the truncated Panx1^1-89 polypeptide. Thus, augmenting extracellular potassium ion concentrations enhances Panx1^1-89-mediated conductance. Moreover, despite the severe truncation, Panx1^1-89 retains sensitivity to most wtPanx1 channel inhibitors. Therefore, Panx1 blockers may be of therapeutic value to combat metastatic cell survival. Our study both provides a mechanism for ATP release from cancer cells and suggests that Panx1^1-89 might aid in the structure-function analysis of Panx1 channels.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635043","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"How to promote co-development between postdocs: a case for peer group meetings.","authors":"Lais Brigliadori Fugio, Tiphaine Douanne, Jane Jardine, Sara Rosińska, Quentin Roux","doi":"10.1111/febs.70067","DOIUrl":"https://doi.org/10.1111/febs.70067","url":null,"abstract":"<p><p>'Who in their right mind would add another meeting to an already overflowing schedule?' This thought crossed our minds when we first discussed the idea of a postdoc peer group meeting. We are five early career researchers working in the same laboratory, each from a different background, shaped as much by our origins, stages in life, and previous research experience. When we started our series of meetings by asking 'What is a postdoc?', we struggled with our own definition of the position, bringing different answers and perspectives. This made us realize that postdoc peer group meetings could be a journey toward open discussion, co-development, and self-discovery.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Critical factors for flow cytometry analysis of the brain.","authors":"Mizuki Sadakata, Ayumu Konno, Akinori Takase, Tetsuhiro Kasamatsu, Takatoshi Iijima, Hirokazu Hirai, Tetsushi Sadakata","doi":"10.1111/febs.70063","DOIUrl":"https://doi.org/10.1111/febs.70063","url":null,"abstract":"<p><p>The brain is difficult to analyze using flow cytometry due to its complex interactions with cells, high lipid content, and high autofluorescence. In this study, we investigated methods to isolate various types of brain cells with high yield and viability. The results showed that protease selection significantly affected the viability of various cell types in the brain. Differences in the developmental stage also affected cell yield and viability. Furthermore, the intensity of autofluorescence differs greatly between various regions of the brain. Additionally, we searched for neuronal indicators capable of identifying a diverse range of neurons. The ratios of various exosomes contained in neurons differ depending on the type of neuronal marker. These results revealed critical factors that must be considered when analyzing various types of brain cells using flow cytometry.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pharmacological manipulation of liver fibrosis progression using novel HDAC6 inhibitors.","authors":"Maria Teresa Borrello, Dusan Ruzic, Hannah Paish, Eleanor Graham, Amy L Collins, Rebecca Scott, Sam Higginbotham, Branko Radovic, Glyn Nelson, David Bulmer, Lee A Borthwick, Stuart M Robinson, Jeremy French, John Moir, Steve A White, Colin Wilson, Sanjay Pandanaboyana, John Hammond, Rohan Thakkar, Wasfi Alrawashdeh, Rodrigo Figueiredo, Milos Petkovic, Katarina Nikolic, Fiona Oakley, Derek A Mann, Jelena Mann","doi":"10.1111/febs.70062","DOIUrl":"https://doi.org/10.1111/febs.70062","url":null,"abstract":"<p><p>Chronic liver injury characterized by unresolved hepatitis leads to fibrosis, potentially progressing to cirrhosis and hepatocellular carcinoma. Effective treatments for halting or reversing liver fibrosis are currently lacking. This study investigates the potential of HDAC6 as a therapeutic target in liver fibrosis. We synthesized two selective HDAC6 inhibitors, DR-3 and FDR2, and assessed their effects on hepatic stellate cell (HSC) activation and liver fibrosis using human precision cut liver slices (hPCLS). Molecular docking, deacetylation inhibition assays, and various cellular assays were employed to evaluate the specificity and anti-fibrotic efficacy of these inhibitors. DR-3 and FDR2 demonstrated high selectivity for HDAC6 over HDAC1, significantly inhibiting HSC activation markers and fibrogenic gene expression. Both inhibitors increased acetylation of α-tubulin and suppressed TGF-β1-induced SMAD signaling in HSCs. In human precision cut liver slices (hPCLS), DR-3 and FDR2 reduced fibrogenic protein levels and collagen deposition. The selective inhibition of HDAC6 by DR-3 and FDR2 effectively reduces HSC activation and fibrogenesis in liver models, supporting further investigation of HDAC6 inhibitors as potential anti-fibrotic therapies.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143627053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rational design of Chim3, a multifunctional peptide carrying a formyl peptide receptor 2 (FPR2) agonist module released by bacterial signal peptidase I (Spase I).","authors":"Samuel R Costa, Bianca O Lira, Gabriel F G Calixto, João B Nunes, Sabrina A Machado, Andreanne G Vasconcelos, Ana L P Lourenço, Thaís C de Sousa, Sónia Gonçalves, André M Murad, Nuno C Santos, José R de S de Almeida Leite, Kelly G Magalhães, Marcelo S Ramada, Guilherme D Brand","doi":"10.1111/febs.70055","DOIUrl":"https://doi.org/10.1111/febs.70055","url":null,"abstract":"<p><p>Membrane-active peptides are useful tools in the design of multifunctional molecules. For example, peptide chimeras may release, after proteolysis of membrane-adsorbed molecules, pharmacologically active fragments. In previous work, Chim2, an antimicrobial peptide composed of a membrane-active module, an enzymatic hydrolysis site, and an agonist moiety for type 2 formyl peptide receptors (FPR2), was conceptualized. Based on Chim2, a peptide named Chim3 was designed, adding a consensus sequence for the bacterial signal peptidase I (Spase I). Spase I is a protease located in an extracytoplasmic face of Gram-positive and Gram-negative bacterial membranes and is essential for protein export. Chim3 was synthesized and its activity as an antimicrobial agent was determined. In addition, Chim3 was incubated with Escherichia coli and Staphylococcus aureus, and peptide hydrolysis products were evaluated by LC-MS/MS. Data demonstrate that Chim3 has potent antimicrobial activity. After incubation with bacteria, Chim3 underwent intense hydrolysis. Proteolysis was detected in the Chim3 Spase I consensus sequence after incubation with both bacteria, and the release of the FPR2 agonist segment was observed. The synthesis of an improved structure of Chim3 with N-methyl tyrosine in the FPR2 agonist segment was performed, resulting in CHIM3Y-NMe. This modification caused significantly higher concentrations of the FPR2 agonist portion arising from the modified peptide after incubation assays with E. coli. The modified FPR2 agonist WK(Y-NMe)M-NH₂ interacted with the mouth region of FPR2 and induced the release of TNF-α and IL-6 in mouse macrophages, making CHIM3Y-NMe an interesting antimicrobial and immunomodulatory molecule for further development aimed at in vivo application.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A steric gate prevents mutagenic dATP incorporation opposite 8-oxo-deoxyguanosine in mitochondrial DNA polymerases.","authors":"Noe Baruch-Torres, Carlos H Trasviña-Arenas, Alexandru Ionut Gilea, Upeksha C Dissanayake, Missael Molina-Jiménez, Paola L García-Medel, Corina Díaz-Quezada, Tiziana Lodi, G Andrés Cisneros, Enrico Baruffini, Luis G Brieba","doi":"10.1111/febs.70064","DOIUrl":"https://doi.org/10.1111/febs.70064","url":null,"abstract":"<p><p>Reactive oxygen species (ROS) generate DNA lesions that alter genome integrity. Among those DNA lesions, 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxodG) is particularly mutagenic. 8-oxodG efficiently incorporates deoxycytidine monophosphate (dCMP) and deoxyadenosine monophosphate (dAMP) via base pairing mediated by its anti and syn conformations, respectively. In family-A DNA polymerases (DNAPs), the amino acids responsible for modulating dCMP or dAMP incorporation across 8-oxodG are located in a determined structural position. Those residues are a conserved tyrosine located at the N terminus of the α-helix O and a nonconserved residue located six amino acids after this conserved tyrosine. In yeast mitochondrial DNAP (DNA-directed DNA polymerase gamma MIP1 [Mip1]), those residues correspond to amino acids Y757 and F763. We hypothesized that the phenyl group of the F763 residue impinges on the syn conformation of 8-oxodG, therefore reducing dAMP misincorporation. Here, we measured dCMP and dAMP incorporation across 8-oxodG using wild-type and F763 Mip1 mutants. Our data suggest that both residue F763 and the universally conserved Y757 assemble a steric gate that obtrudes the 8-oxodG(syn) conformation. As the human orthologue of Mip1, DNA polymerase gamma (HsPolγ) or DNAP γ, also harbors phenylalanine at the corresponding position to Mip1-F763, the steric gate mechanism might similarly be responsible for controlling HsPolγ's fidelity when tolerating 8-oxodG lesions.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607219","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Caspase-1/11 controls Zika virus replication in astrocytes by inhibiting glycolytic metabolism.","authors":"Ingrid S de Farias, Guilherme Ribeiro, Isaú H Noronha, Victoria Weise L Lucena, Jean P S Peron, Pedro M Moraes-Vieira, Jose C Alves-Filho, Karina R Bortoluci","doi":"10.1111/febs.70061","DOIUrl":"https://doi.org/10.1111/febs.70061","url":null,"abstract":"<p><p>Zika virus (ZIKV) poses a significant threat due to its association with severe neurological complications, particularly during pregnancy. Although viruses exhibit tropism for neural cells, including astrocytes, the role of these cells in controlling ZIKV replication remains unclear. In this study, we demonstrated that ZIKV induces caspase-1 activation in primary astrocytes despite the absence of classical signs of inflammasome activation. Caspase-1 and caspase-11 double knockout (caspase-1/11^-/-) astrocytes exhibit heightened permissiveness to viral replication, accompanied by overactivation of glycolytic metabolism. Inhibition of glycolysis reversed the susceptibility of caspase-1/11^-/- astrocytes to ZIKV infection. Protein network analysis revealed mammalian target of rapamycin complex (mTORC) as a link between proteins involved in glycolysis and caspase-1, and mTORC inhibition also suppressed viral replication. Furthermore, we found that the impact of caspase-1/11 on astrocytes depends on the regulation of pyruvate transport to mitochondria for viral replication. Overall, our findings elucidate a caspase-1/11-dependent microbicidal mechanism in astrocytes that involves the mTORC/glycolytic pathway/pyruvate axis, providing insights into potential therapeutic targets for ZIKV infection.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The deubiquitinase inhibitor WP1130 drives nuclear aggregation and reactivation of mutant p53 for selective cancer cell targeting.","authors":"Swapnil Oak, Onkar Karajgikar, Nikhil Gadewal, Prasad Sulkshane, Tripti Verma, Sanjay Gupta, Tanuja Teni","doi":"10.1111/febs.70036","DOIUrl":"https://doi.org/10.1111/febs.70036","url":null,"abstract":"<p><p>Mutations in the TP53 gene may lead to the loss of its tumor suppressor function and the acquisition of oncogenic properties. The enhanced stability of mutant p53 (mutp53) is one of the pivotal factors for its oncogenic functions, rendering proteins implicated in mutp53 stabilization as promising targets for therapeutic intervention. Although deubiquitinases (DUBs) are commonly deregulated in various cancers, their specific impact on mutp53 stabilization remains largely unexplored. In this study, we demonstrated the involvement of DUBs-USP5 and USP9X in-enhancing mutp53 stability while revealing the effects of DUB inhibitor WP1130 in selectively destabilizing different p53 mutants in cancer cells of various origins. Mechanistically, WP1130 induced mutp53 ubiquitination and nuclear aggregation, resulting in its partitioning to the detergent-insoluble fraction. Moreover, combined treatment with the proteasome inhibitor augmented mutp53 accumulation in this fraction, indicating proteasomal degradation of these aggregates. Interestingly, WP1130 did not alter the stability or induce aggregation of WTp53 protein, suggesting its selective targeting of mutp53. Furthermore, WP1130 disrupted the interaction of mutp53 with HSP40 and HSP90 while promoting its association with ubiquitin ligase CHIP, thereby facilitating mutp53 destabilization. Notably, WP1130 reactivated mutp53 via induction of a wild-type-like p53 conformation, upregulating its downstream effectors and inducing apoptosis, possibly due to its targeted binding near the mutation site, as suggested by our in silico analysis. These findings highlight the roles of USP9X and USP5 in mutp53 stabilization and underscore the therapeutic potential of DUB inhibitor WP1130 for the selective targeting of mutp53-expressing cancer cells.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Derivatives of MOPS: promising scaffolds for SARS coronaviruses Macro domain-targeted inhibition.","authors":"Oney Ortega Granda, Karine Alvarez, Benjamin Morin, Bruno Canard, François Ferron, Nadia Rabah","doi":"10.1111/febs.70039","DOIUrl":"https://doi.org/10.1111/febs.70039","url":null,"abstract":"<p><p>The severe acute respiratory syndrome coronavirus (SARS-CoV/CoV-2) genome encodes 16 non-structural proteins (nsps), which coordinate cell remodeling, virus replication and participate in viral evasion. Notably, nsp3 contains a protein module termed Macro domain, which carries IFN antagonist activity that interferes with host innate immunity response. This domain is able to bind and hydrolyze ADP-ribose derivatives. This activity is correlated to viral escape and thus makes Macro domains a valuable therapeutic target. In the present paper, we report a SARS-CoV Macro domain structure in complex with a MOPS molecule. Based on our structural data, molecular docking was performed on a set of MOPS analogs in the ADP-ribose binding pocket. We present an ELISA-based assay to select hits based on the inhibition of recombinant SARS-CoV/CoV-2 Macro domain-ADP-ribose complex formation. Among the tested analogs, MOPSO and CAPSO are the more efficient in inhibiting ADP-ribose-binding. Structural analysis of these molecules in the ADP-ribose pocket reveals potential interactions with amino acid residues involved in the coordination of ADP-ribose. Overall, these findings suggest that MOPSO and CAPSO bear potential to be used as a scaffold for the design of Macro domain-specific inhibitors.</p>","PeriodicalId":94226,"journal":{"name":"The FEBS journal","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143607181","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}