IF 6.4 1区生物学

eLife Pub Date : 2025-07-08 DOI: 10.7554/eLife.102068

Renaud Prevel, Erwan Pernet, Kim A Tran, Abderrahmane Sadek, Mina Sadeghi, Elizabeth Lapshina, Leonardo F Jurado, Arnold S Kristof, Mohieddine Moumni, Jeremie Poschmann, Maziar Divangahi

{"title":"β-Glucan reprograms alveolar macrophages via neutrophil/IFNγ axis in a murine model of lung injury.","authors":"Renaud Prevel, Erwan Pernet, Kim A Tran, Abderrahmane Sadek, Mina Sadeghi, Elizabeth Lapshina, Leonardo F Jurado, Arnold S Kristof, Mohieddine Moumni, Jeremie Poschmann, Maziar Divangahi","doi":"10.7554/eLife.102068","DOIUrl":"10.7554/eLife.102068","url":null,"abstract":"Alveolar macrophages (AMs) reside in the lower airways and play a crucial role in lung health and response to sterile inflammation and infections. AMs possess remarkable adaptability to different environmental challenges that can persist through their memory capacity (trained immunity). β-Glucan has been characterized as a potent inducer of central trained immunity by reprogramming haematopoietic stem cells in the bone marrow. In the present study, we show that systemic administration of β-glucan in mice induces peripheral trained immunity by reprogramming AMs in the lungs, in a Dectin1-independent manner. We furthermore demonstrate that AM reprogramming at both the transcriptional and metabolic levels exacerbate lung injury following bacterial (lipopolysaccharide) or viral (polyI:C) challenges via a neutrophil/IFN-γ-dependent manner. These findings identify an additional facet of β-glucan in trained immunity involving AM reprogramming and shed light on the potential detrimental effects of trained immunity.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12237401/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144583417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Diverse prey capture strategies in teleost larvae. 硬骨鱼幼虫捕食策略的多样性。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-08 DOI: 10.7554/eLife.98347

Duncan S Mearns, Sydney A Hunt, Martin W Schneider, Ash V Parker, Manuel Stemmer, Herwig Baier

{"title":"Diverse prey capture strategies in teleost larvae.","authors":"Duncan S Mearns, Sydney A Hunt, Martin W Schneider, Ash V Parker, Manuel Stemmer, Herwig Baier","doi":"10.7554/eLife.98347","DOIUrl":"10.7554/eLife.98347","url":null,"abstract":"Animal behavior is adapted to the sensory environment in which it evolved, while also being constrained by physical limits, evolutionary history, and developmental trajectories. The hunting behavior of larval zebrafish (Danio rerio), a cyprinid native to streams in Eastern India, has been well characterized. However, it is unknown if the complement and sequence of movements employed during prey capture by zebrafish is universal across freshwater teleosts. Here, we explore the syntax of prey capture behavior in larval fish belonging to the clade Percomorpha, whose last common ancestor with cyprinids lived ~240 mya. We compared the behavior of four cichlid species from Lake Tanganyika endemic to deep benthic parts of the lake (Lepidiolamprologus attenuatus, Lamprologus ocellatus, and Neolamprologus multifasciatus) or inhabiting rivers (Astatotilapia burtoni) with that of medaka (Oryzias latipes), a fish found in rice paddies in East Asia. Using high-speed videography and neural networks, we tracked eye movements and extracted swim kinematics during hunting from larvae of these five species. Notably, we found that the repertoire of hunting movements of cichlids is broader than that of zebrafish, but shares basic features, such as eye convergence, positioning of prey centrally in the binocular visual field, and discrete prey capture bouts, including two kinds of capture strikes. In contrast, medaka swim continuously, track the prey monocularly without eye convergence, and position prey laterally before capturing them with a side swing. This configuration of kinematic motifs suggests that medaka may judge distance to prey predominantly by motion parallax, while cichlids and zebrafish may mainly use binocular visual cues. Together, our study documents the diversification of locomotor and oculomotor adaptations among hunting teleost larvae.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12237402/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144583414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Unraveling CRP/cAMP-mediated metabolic regulation in Escherichia coli persister cells. 揭示CRP/ camp介导的大肠杆菌持久性细胞代谢调节。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-08 DOI: 10.7554/eLife.99735

Han G Ngo, Sayed Golam Mohiuddin, Aina Ananda, Mehmet Orman

{"title":"Unraveling CRP/cAMP-mediated metabolic regulation in Escherichia coli persister cells.","authors":"Han G Ngo, Sayed Golam Mohiuddin, Aina Ananda, Mehmet Orman","doi":"10.7554/eLife.99735","DOIUrl":"10.7554/eLife.99735","url":null,"abstract":"A substantial gap persists in our comprehension of how bacterial metabolism undergoes rewiring during the transition to a persistent state. Also, it remains unclear which metabolic mechanisms become indispensable for persister cell survival. To address these questions, we directed our efforts towards persister cells in Escherichia coli that emerge during the late stationary phase. These cells have been recognized for their exceptional resilience and are commonly believed to be in a dormant state. Our results indicate that the global metabolic regulator Crp/cAMP redirects the metabolism of these antibiotic-tolerant cells from anabolism to oxidative phosphorylation. Although our data demonstrates that persisters exhibit a reduced metabolic rate compared to rapidly growing exponential-phase cells, their survival still relies on energy metabolism. Extensive genomic-level analyses of metabolomics, proteomics, and single-gene deletions consistently highlight the critical role of energy metabolism, specifically the tricarboxylic acid (TCA) cycle, electron transport chain (ETC), and ATP synthase, in sustaining persister levels within cell populations. Altogether, this study provides much-needed clarification regarding the role of energy metabolism in antibiotic tolerance and highlights the importance of using a multipronged approach at the genomic level to obtain a broader picture of the metabolic state of persister cells.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12237405/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590717","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Homophilic wiring principles underpin neuronal network topology in vitro. 同质布线原理支持体外神经网络拓扑结构。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-08 DOI: 10.7554/eLife.85300

Danyal Akarca, Alexander W E Dunn, Philipp J Hornauer, Silvia Ronchi, Michele Fiscella, Congwei Wang, Marco Terrigno, Ravi Jagasia, Petra E Vértes, Susanna B Mierau, Ole Paulsen, Stephen J Eglen, Andreas Hierlemann, Duncan E Astle, Manuel Schröter

{"title":"Homophilic wiring principles underpin neuronal network topology in vitro.","authors":"Danyal Akarca, Alexander W E Dunn, Philipp J Hornauer, Silvia Ronchi, Michele Fiscella, Congwei Wang, Marco Terrigno, Ravi Jagasia, Petra E Vértes, Susanna B Mierau, Ole Paulsen, Stephen J Eglen, Andreas Hierlemann, Duncan E Astle, Manuel Schröter","doi":"10.7554/eLife.85300","DOIUrl":"https://doi.org/10.7554/eLife.85300","url":null,"abstract":"Economic efficiency has been a popular explanation for how networks self-organize within the developing nervous system. However, the precise nature of the economic negotiations governing this putative organizational principle remains unclear. Here, we address this question further by combining large-scale electrophysiological recordings, to characterize the functional connectivity of developing neuronal networks in vitro, with a generative modeling approach capable of simulating network formation. We find that the best fitting model uses a homophilic generative wiring principle in which neurons form connections to other neurons which are spatially proximal and have similar connectivity patterns to themselves. Homophilic generative models outperform more canonical models in which neurons wire depending upon their spatial proximity either alone or in combination with the extent of their local connectivity. This homophily-based mechanism for neuronal network emergence accounts for a wide range of observations that are described, but not sufficiently explained, by traditional analyses of network topology. Using rodent and human neuronal cultures, we show that homophilic generative mechanisms can accurately recapitulate the topology of emerging cellular functional connectivity, representing an important wiring principle and determining factor of neuronal network formation in vitro.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"14 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144583420","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}

引用次数: 0

Forward genetics in C. elegans reveals genetic adaptations to polyunsaturated fatty acid deficiency. 秀丽隐杆线虫的正向遗传学揭示了对多不饱和脂肪酸缺乏的遗传适应。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-08 DOI: 10.7554/eLife.104181

Delaney Kaper, Uroš Radović, Per-Olof Bergh, August Qvist, Marcus Henricsson, Jan Borén, Marc Pilon

{"title":"Forward genetics in C. elegans reveals genetic adaptations to polyunsaturated fatty acid deficiency.","authors":"Delaney Kaper, Uroš Radović, Per-Olof Bergh, August Qvist, Marcus Henricsson, Jan Borén, Marc Pilon","doi":"10.7554/eLife.104181","DOIUrl":"10.7554/eLife.104181","url":null,"abstract":"Polyunsaturated fatty acids (PUFAs) are essential for mammalian health and function as membrane fluidizers and precursors for signaling lipids, though the primary essential function of PUFAs within organisms has not been established. Unlike mammals who cannot endogenously synthesize PUFAs, C. elegans can de novo synthesize PUFAs starting with the Δ12 desaturase FAT-2, which introduces a second double bond to monounsaturated fatty acids to generate the PUFA linoleic acid. FAT-2 desaturation is essential for C. elegans survival since fat-2 null mutants are non-viable; the near-null fat-2(wa17) allele synthesizes only small amounts of PUFAs and produces extremely sick worms. Using fluorescence recovery after photobleaching (FRAP), we found that the fat-2(wa17) mutant has rigid membranes and can be efficiently rescued by dietarily providing various PUFAs, but not by fluidizing treatments or mutations. With the aim of identifying mechanisms that compensate for PUFA-deficiency, we performed a forward genetics screen to isolate novel fat-2(wa17) suppressors and identified four internal mutations within fat-2 and six mutations within the HIF-1 pathway. The suppressors increase PUFA levels in fat-2(wa17) mutant worms and additionally suppress the activation of the daf-16, UPRer and UPRmt stress response pathways that are active in fat-2(wa17) worms. We hypothesize that the six HIF-1 pathway mutations, found in egl-9, ftn-2, and hif-1, all converge on raising Fe2+ levels and in this way boost desaturase activity, including that of the fat-2(wa17) allele. We conclude that PUFAs cannot be genetically replaced and that the only genetic mechanism that can alleviate PUFA-deficiency do so by increasing PUFA levels.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12237404/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144590714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The molecular logic of Gtr1/2- and Pib2-dependent TORC1 regulation in budding yeast. 出芽酵母中Gtr1/2-和pib2依赖性TORC1调控的分子逻辑。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-07 DOI: 10.7554/eLife.94628

Jacob H Cecil, Cristina M Padilla, Austin A Lipinski, Paul Langlais, Xiangxia Luo, Andrew P Capaldi

引用次数: 0

Magnetically steered cell therapy for reduction of intraocular pressure as a treatment strategy for open-angle glaucoma. 磁导向细胞治疗开角型青光眼降低眼压的研究。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-07 DOI: 10.7554/eLife.103256

M Reza Bahranifard, Jessica Chan, A Thomas Read, Guorong Li, Lin Cheng, Babak N Safa, Seyed Mohammad Siadat, Anamik Jhunjhunwala, Hans E Grossniklaus, Stanislav Y Emelianov, W Daniel Stamer, Markus H Kuehn, C Ross Ethier

{"title":"Magnetically steered cell therapy for reduction of intraocular pressure as a treatment strategy for open-angle glaucoma.","authors":"M Reza Bahranifard, Jessica Chan, A Thomas Read, Guorong Li, Lin Cheng, Babak N Safa, Seyed Mohammad Siadat, Anamik Jhunjhunwala, Hans E Grossniklaus, Stanislav Y Emelianov, W Daniel Stamer, Markus H Kuehn, C Ross Ethier","doi":"10.7554/eLife.103256","DOIUrl":"10.7554/eLife.103256","url":null,"abstract":"Trabecular meshwork (TM) cell therapy has been proposed as a next-generation treatment for elevated intraocular pressure (IOP) in glaucoma, the most common cause of irreversible blindness. Using a magnetic cell steering technique with excellent efficiency and tissue-specific targeting, we delivered two types of cells into a mouse model of glaucoma: either human adipose-derived mesenchymal stem cells (hAMSCs) or induced pluripotent cell derivatives (iPSC-TM cells). We observed a 4.5 [3.1, 6.0] mmHg or 27% reduction in intraocular pressure (IOP) for 9 months after a single dose of only 1500 magnetically steered hAMSCs, explained by increased outflow through the conventional pathway and associated with a higher TM cellularity. iPSC-TM cells were also effective, but less so, showing only a 1.9 [0.4, 3.3] mmHg or 13% IOP reduction and increased risk of tumorigenicity. In both cases, injected cells remained detectable in the iridocorneal angle 3 weeks post-transplantation. Based on the locations of the delivered cells, the mechanism of IOP lowering is most likely paracrine signaling. We conclude that magnetically steered hAMSC cell therapy has potential for long-term treatment of ocular hypertension in glaucoma.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12234007/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144575082","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The ESCRT protein CHMP5 restricts bone formation by controlling endolysosome-mitochondrion-mediated cell senescence. ESCRT蛋白CHMP5通过控制内溶酶体-线粒体介导的细胞衰老来限制骨形成。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-07 DOI: 10.7554/eLife.101984

Fan Zhang, Yuan Wang, Luyang Zhang, Chunjie Wang, Deping Chen, Haibo Liu, Ren Xu, Cole M Haynes, Jae-Hyuck Shim, Xianpeng Ge

{"title":"The ESCRT protein CHMP5 restricts bone formation by controlling endolysosome-mitochondrion-mediated cell senescence.","authors":"Fan Zhang, Yuan Wang, Luyang Zhang, Chunjie Wang, Deping Chen, Haibo Liu, Ren Xu, Cole M Haynes, Jae-Hyuck Shim, Xianpeng Ge","doi":"10.7554/eLife.101984","DOIUrl":"10.7554/eLife.101984","url":null,"abstract":"The dysfunction of the cellular endolysosomal pathway, such as in lysosomal storage diseases, can cause severe musculoskeletal disorders. However, how endolysosomal dysfunction causes musculoskeletal abnormalities remains poorly understood, limiting therapeutic options. Here, we report that CHMP5, a member of the endosomal sorting complex required for transport (ESCRT)-III protein family, is essential to maintain the endolysosomal pathway and regulate bone formation in osteogenic lineage cells. Genetic ablation of Chmp5 in mouse osteogenic cells increases bone formation in vivo and in vitro. Mechanistically, Chmp5 deletion causes endolysosomal dysfunction by decreasing the VPS4A protein, and CHMP5 overexpression is sufficient to increase the VPS4A protein. Subsequently, endolysosomal dysfunction disturbs mitochondrial functions and increases mitochondrial ROS, ultimately resulting in skeletal cell senescence. Senescent skeletal cells cause abnormal bone formation by combining cell-autonomous and paracrine actions. Importantly, the elimination of senescent cells using senolytic drugs can alleviate musculoskeletal abnormalities in Chmp5 conditional knockout mice. Therefore, our results show that cell senescence represents an underpinning mechanism and a therapeutic target for musculoskeletal disorders caused by the aberrant endolysosomal pathway, such as in lysosomal storage diseases. These results also uncover the function and mechanism of CHMP5 in the regulation of cell senescence by affecting the endolysosomal-mitochondrial pathway.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"13 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12234009/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144575083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Highly sensitive in vivo detection of dynamic changes in enkephalins following acute stress in mice. 小鼠急性应激后脑啡肽动态变化的高灵敏度体内检测。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-07 DOI: 10.7554/eLife.91609

Marwa O Mikati, Petra Erdmann-Gilmore, Rose Connors, Sineadh M Conway, Jim Malone, Justin Woods, Robert W Sprung, Reid R Townsend, Ream Al-Hasani

{"title":"Highly sensitive in vivo detection of dynamic changes in enkephalins following acute stress in mice.","authors":"Marwa O Mikati, Petra Erdmann-Gilmore, Rose Connors, Sineadh M Conway, Jim Malone, Justin Woods, Robert W Sprung, Reid R Townsend, Ream Al-Hasani","doi":"10.7554/eLife.91609","DOIUrl":"10.7554/eLife.91609","url":null,"abstract":"Enkephalins are opioid peptides that modulate analgesia, reward, and stress. In vivo detection of enkephalins remains difficult due to transient and low endogenous concentrations and inherent sequence similarity. To begin to address this, we previously developed a system combining in vivo optogenetics with microdialysis and a highly sensitive mass spectrometry-based assay to measure opioid peptide release in freely moving rodents (Al-Hasani et al., 2018, eLife). Here, we show improved detection resolution and stabilization of enkephalin detection, which allowed us to investigate enkephalin release during acute stress. We present an analytical method for real-time, simultaneous detection of Met- and Leu-enkephalin (Met-Enk and Leu-Enk) in the mouse nucleus accumbens shell (NAcSh) after acute stress. We confirm that acute stress activates enkephalinergic neurons in the NAcSh using fiber photometry and that this leads to the release of Met- and Leu-Enk. We also demonstrate the dynamics of Met- and Leu-Enk release as well as how they correlate to one another in the ventral NAc shell, which was previously difficult due to the use of approaches that relied on mRNA transcript levels rather than posttranslational products. This approach increases spatiotemporal resolution, optimizes the detection of Met-Enk through methionine oxidation, and provides novel insight into the relationship between Met- and Leu-Enk following stress.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"12 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12234004/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144575085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The C-terminus of the multi-drug efflux pump EmrE prevents proton leak by gating transport. 多药外排泵EmrE的c端通过门控输运防止质子泄漏。

IF 6.4 1区生物学

eLife Pub Date : 2025-07-07 DOI: 10.7554/eLife.105525

Merissa Brousseau, Da Teng, Nathan E Thomas, Gregory A Voth, Katherine A Henzler-Wildman

{"title":"The C-terminus of the multi-drug efflux pump EmrE prevents proton leak by gating transport.","authors":"Merissa Brousseau, Da Teng, Nathan E Thomas, Gregory A Voth, Katherine A Henzler-Wildman","doi":"10.7554/eLife.105525","DOIUrl":"10.7554/eLife.105525","url":null,"abstract":"The model multi-drug efflux pump from Escherichia coli, EmrE, can perform multiple types of transport leading to different biological outcomes, conferring resistance to some drug substrates and enhancing susceptibility to others. While transporters have traditionally been classified as antiporters, symporters, or uniporters, there is growing recognition that some transporters may exhibit mixed modalities. This raises new questions about their regulation and mechanism. Here, we show that the C-terminal tail of EmrE acts as a secondary gate, preventing proton leak in the absence of drug. Substrate binding unlocks this gate, allowing transport to proceed. Truncation of the C-terminal tail (∆107-EmrE) leads to altered pH regulation of alternating access, an important kinetic step in the transport cycle, as measured by NMR. ∆107-EmrE has increased proton leak in proteoliposomes, and bacteria expressing this mutant have reduced growth. Molecular dynamics simulations of ∆107-EmrE show the formation of a water wire from the open face of the transporter to the primary binding site in the core, facilitating proton leak. In WT-EmrE, the C-terminal tail forms specific interactions that block the formation of the water wire. Together, these data strongly support the C-terminus of EmrE acting as a secondary gate that regulates access to the primary binding site.","PeriodicalId":11640,"journal":{"name":"eLife","volume":"14 ","pages":""},"PeriodicalIF":6.4,"publicationDate":"2025-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12234006/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144583431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

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