ATP Synthase Subunit Research Articles

Comprehensive analysis of the succinylome in Vero cells infected with peste des petits ruminants virus Nigeria 75/1 vaccine strain.

Peste des petits ruminants virus (PPRV) is currently the only member of the Morbillivirus caprinae species within the genus Morbillivirus of the family Paramyoxviridae. PPRV causes a highly contagious disease in small ruminants, especially goats and sheep. Succinylation is a newly identified and conserved modification and plays an important role in host cell response to pathogen infection. However, the extent and function of succinylation in Vero cells during PPRV infection remains unknown. In this study, a global profile of the succinylome in Vero cells infected with PPRV Nigeria 75/1 vaccine strain (PPRVvac) was performed by dimethylation labeling-based quantitative proteomics analysis. A total of 2633 succinylation sites derived from 823 proteins were quantified. The comparative analysis of differentially succinylated sites revealed that 228 down-regulated succinylation sites on 139 proteins and 44 up-regulated succinylation sites on 38 proteins were significantly modified in response to PPRVvac infection, seven succinylation motifs were identified. GO classification indicated that the differentially succinylated proteins (DSuPs) mainly participated in cellular respiration, biosynthetic process and transmembrane transporter activity. KEGG pathway analysis indicated that DSuPs were related to protein processing in the endoplasmic reticulum. Protein-protein interaction networks of the identified proteins provided further evidence that various ATP synthase subunits and carbon metabolism were modulated by succinylation, while the overlapped proteins between succinylation and acetylation are involved in glyoxylate and dicarboxylate metabolism. The findings of the present study provide the first report of the succinylome in Vero cells infected with PPRVvac and provided a foundation for investigating the role of succinylation alone and its overlap with acetylation in response to PPRVvac.

ATP5F1D: A central player in mitochondrial energy metabolism and its implications in disease

ATP synthase f1 subunit delta (ATP5F1D, previously APT5D), a subunit of mitochondrial ATP synthase, plays a pivotal role in ATP production and maintaining cellular energy balance. This study explores ATP5F1D’s involvement in energy metabolism and its implications in disease through a comprehensive review of approximately 100 articles published over the past two decades. Findings indicate that ATP5F1D is closely associated with metabolic adaptation, oxidative stress, and apoptotic regulation, with its dysregulation linked to pathological conditions such as tumor energy reprogramming and cardiovascular energy deficits. As a critical biomarker and therapeutic target, ATP5D holds promise for advancing the understanding of mitochondrial biology and metabolic diseases.

Transcriptome Analysis Reveals Key Genes Involved in the Response of Triticum urartu to Boron Toxicity Stress

The domestication and breeding of wheat genotypes through the years has led to the loss in their genetic variation, making them more prone to different abiotic stresses. Boron (B) toxicity is one of the stresses decreasing the wheat cultivars’ yield in arid and semi-arid regions around the world. Wild wheat progenitors, such as Triticum urartu Thumanian ex Gandilyan, possess a broader gene pool that harbors several genes conferring tolerance to various biotic and abiotic stresses. Unfortunately, T. urartu is not well-explored at the molecular level for its tolerance towards B toxicity in soil. In this study, for the first time, we compared the transcriptomic changes in the leaves of a high B-tolerant T. urartu genotype, PI662222, grown in highly toxic B (10 mM B in the form of boric acid) with the ones grown in the control (3.1 μM B) treatment in hydroponic conditions. The obtained results suggest that several mechanisms are involved in regulating the response of the studied T. urartu genotype toward B toxicity. All the growth parameters of T. urartu genotype, including root–shoot length, root fresh weight, and root–shoot dry weight, were less affected by high boron (10 mM) as compared to the boron-tolerant bread wheat cultivar. With a significant differential expression of 654 genes, 441 and 213 genes of T. urartu genotype were down- and upregulated, respectively, in the PI662222 leaves in high B in comparison to the control treatment. While key upregulated genes included those encoding RNA polymerase beta subunit (chloroplast), ATP synthase subunit gamma, chloroplastic, 60S ribosomal protein, and RNA-binding protein 12-like, the main downregulated genes included those encoding photosystem II protein D, ribulose bisphosphate carboxylase small subunit, and peroxidase 2-like. Interestingly, both Gene Ontology enrichment and KEGG pathways emphasized the possible involvement of the genes related to the photosynthetic process and apparatus in the high B tolerance of the T. urartu genotype. The further functional characterization of the identified potential T. urartu genes will facilitate their utilization in crop improvement programs for B toxicity stress.

Long Non-Coding RNA LOC113219358 Regulates Immune Responses in Apis mellifera Through Protein Interactions.

Long non-coding RNAs (lncRNAs) are emerging as critical regulators in honeybee physiology, influencing development, behavior, and stress responses. This study investigates the role of lncRNA LOC113219358 in the immune response and neurophysiological regulation of Apis mellifera brains. Using RNA interference (RNAi) and RNA sequencing (RNA-seq), we demonstrate that silencing lncLOC113219358 significantly alters the expression of 162 mRNA transcripts, including genes associated with detoxification, energy metabolism, and neuronal signaling. Functional enrichment analysis revealed involvement in neuropeptide signaling, ATP synthesis, and oxidative phosphorylation pathways. Acetylcholinesterase (AChE), Glutathione-S-transferase (GST) and cytochrome P450 (CYP450) activities were significantly downregulated with 48 h of RNAi treatment. Additionally, RNA pull-down assays identified 113 proteins interacting with lncLOC113219358, including ATP synthase subunits, heat shock proteins, and major royal jelly proteins, suggesting its role in cellular stress responses and neural activity modulation. These findings provide mechanistic insights into how lncLOC113219358 mediates honeybee responses to environmental stressors, contributing to our understanding of lncRNA-regulated neural and immune functions in pollinators.

Adenine nucleotide translocator and ATP synthase cooperate in mediating the mitochondrial permeability transition.

The permeability transition (PT) is a permeability increase of the mitochondrial inner membrane causing mitochondrial swelling in response to matrix Ca2+. The PT is mediated by regulated channel(s), the PT pore(s) (PTP), which can be generated by at least two components, adenine nucleotide translocator (ANT) and ATP synthase. Whether these provide independent permeation pathways remains to be established. Here, we assessed the contribution of ANT to the PT based on the effects of the selective ANT inhibitors atractylate (ATR) and bongkrekate (BKA), which trigger and inhibit channel formation by ANT, respectively. BKA partially inhibited Ca2+-dependent PT and did not prevent the inducing effect of phenylarsine oxide, which was still present in mouse embryonic fibroblasts deleted for all ANT isoforms. The contribution of ANT to the PT emerged at pH 6.5 (a condition that inhibits ATP synthase channel opening) in the presence of ATR, which triggered mitochondrial swelling and elicited currents in patch-clamped mitoplasts. Unexpectedly, ANT-dependent PT at pH 6.5 could also be stimulated by benzodiazepine-423 [a selective ligand of the oligomycin sensitivity conferral protein (OSCP) subunit of ATP synthase], suggesting that the ANT channel is regulated by the peripheral stalk of ATP synthase. In keeping with docking simulations, ANT could be co-immunoprecipitated with ATP synthase subunits c and g, and oligomycin (which binds adjacent c subunits) decreased the association of ANT with subunit c. These results reveal a close cooperation between ANT and ATP synthase in the PT and open new perspectives in the study of this process. KEY POINTS: We have assessed the relative role of adenine nucleotide translocator (ANT) and ATP synthase in generating the mitochondrial permeability transition (PT). At pH 7.4, bongkrekate had little effect on Ca2+-dependent PT, and did not prevent the inducing effect of phenylarsine oxide, which was still present in mouse embryonic fibroblasts deleted for all ANT isoforms. The contribution of ANT emerged at pH 6.5 (which inhibits ATP synthase channel opening) in the presence of atractylate, which triggered mitochondrial swelling and elicited currents in patch-clamped mitoplasts. Benzodiazepine-423, a selective ligand of the oligomycin sensitivity conferral protein subunit of ATP synthase, stimulated ANT-dependent PT at pH 6.5, suggesting that the ANT channel is regulated by the peripheral stalk of ATP synthase. ANT could be co-immunoprecipitated with ATP synthase subunits c and g; oligomycin, which binds adjacent c subunits, decreased the association with subunit c, in keeping with docking simulations.

Breaking the energy chain: importance of ATP synthase in Mycobacterium tuberculosis and its potential as a drug target.

Unveiling novel pathways for drug discovery forms the foundation of a new era in the combat against tuberculosis. The discovery of a novel drug, bedaquiline, targeting mycobacterial ATP synthase highlighted the targetability of the energy metabolism pathway. The significant potency of bedaquiline against heterogeneous population of Mycobacterium tuberculosis marks ATP synthase as an important complex of the electron transport chain. This review focuses on the importance and unique characteristics of mycobacterial ATP synthase. Understanding these distinctions enables the targeting of ATP synthase subunits for drug discovery, without aiming at the mammalian counterpart. Furthermore, a brief comparison of the structural differences between mycobacterial and mitochondrial ATP synthase is discussed. Being a complex multi-subunit protein, ATP synthase offers multiple sites for potential inhibitors, including the a, c, ε, γ, and δ subunits. Inhibitors targeting these subunits are critically reviewed, providing insight into the design of better and more potent chemical entities with the potential for effective treatment regimens.

High Glucose Inhibits O-GlcNAc Transferase Translocation in Early Osteoblast Differentiation by Altering Protein Phosphatase 2A Activity.

Our previous study revealed a link between O-GlcNAc transferase (OGT) localization and protein phosphatase 2A (PP2A) activity in osteoblast. Given the association of PP2A downregulation with osteoblast differentiation, we hypothesized that OGT localization changes during this process. We examined OGT localization in MC3T3-E1 cells undergoing differentiation under normal and high glucose conditions. Changes in PP2A activity were followed by alterations in OGT localization. Organ culture of calvaria revealed similar OGT localization changes in bone-surrounding osteoblasts near the suture area. Furthermore, the levels of O-GlcNAc modification in various proteins including Runt-related transcription factor 2, Osterix, and ATP synthase subunit alpha (ATP5A) were shifted in parallel with OGT translocation. These findings suggest a regulatory role of OGT, under the influence of PP2A, during osteoblast differentiation.

Catestatin improves heart metabolic flexibility by promoting mitochondrial structure and function.

Hypertension, a major cause of cardiomyopathy, is one of the most critical risk factors for heart failure and mortality worldwide. Loss of metabolic flexibility of cardiomyocytes is one of the major causes of heart failure. Although Catestatin (CST) treatment is known to be both hypotensive and cardioprotective, its effect on cardiac metabolism is unknown. In this study, we undertook a transcriptomic approach to identify differentially expressed genes that were filtered using Boolean implication relationships to develop a model of gene regulation in saline or CST-supplemented CST knockout (CST-KO) mice. The analysis revealed a set of gene signatures (fibroblast, cardiomyocyte, and macrophage) rescued after CST supplemented CST-KO mice compared to wild-type. Furthermore, we independently validated these gene signature models using publicly available patient datasets. Since the gene signature includes genes related to glucose, fatty acid metabolism, and mitochondrial function, we assessed the glucose and fatty acid uptake after CST treatment. We found that CST treatment can restore the cardiac metabolic inflexibility in CST-KO heart due to the metabolic shift of glucose utilization to fatty acid as energy source. Binding studies after immunoprecipitation and mass spectrometry revealed CST binding with ATP synthase, supported by molecular simulation and computational modeling that predicted CST binding to α/β subunit of ATP synthase. Colocalization of CST with mitochondria and increased mitochondrial membrane potential and ATP production upon CST treatment in neonatal cardiomyocytes further exhibit CST as a key regulator of cardiac metabolism and mitochondrial function.

Integrating structure-guided and fragment-based inhibitor design to combat bedaquiline resistant Mycobacterium tuberculosis: a molecular dynamics study.

The first FDA approved, MDR-TB inhibitory drug bedaquiline (BDQ), entraps the c-ring of the proton-translocating F0 region of enzyme ATP synthase of Mycobacterium tuberculosis, thus obstructing successive ATP production. Present-day BDQ-resistance has been associated with cardiotoxicity and mutation(s) in the atpE gene encoding the c subunit of ATP synthase (ATPc) generating five distinct ATPc mutants: Ala63→Pro, Ile66→Met, Asp28→Gly, Asp28→Val and Glu61→Asp. We created three discrete libraries, first by repurposing bedaquiline via scaffold hopping approach, second one having natural plant compounds and the third being experimentally derived analogues of BDQ to identify one drug candidate that can inhibit ATPc activity more efficiently with less toxic properties. For this purpose, we adopted techniques like molecular dynamics simulation, virtual screening, PCA, DCCM, binding affinity analysis to gauge structure-function relationship of the L136-ATPc complexes. L136 was found to induce a distinguishable conformational change in the bound ATPc which captivated the c9 rotor ring. L136 displays a binding free energy of -57.294, -59.027, -57.273, -58.726, -55.889 and -58.651 kcal/mol for ATPc_WT and the five respective mutants. The pIC50 value for the L136 ligand for the same proteins was unveiled to be 6.760, 7.285, 6.898, 7.222, 6.987 and 7.687. Moreover, L136 exhibited a strong ADMET profile. Furthermore, we discovered that the change in the hydrophobic platform in ATPc mutants hinders BDQ binding, which is overcome by L136, ensuring efficient binding and providing an assessment of L136's mechanism of ATPc inhibition. L136 provides a scope for invivo test for future clinical drug trials.

Causal relationship between mitochondrial function and delirium: a bidirectional two-sample Mendelian randomisation analysis.

Previous studies have suggested a potential link between delirium and mitochondrial function. Consequently, this Mendelian randomisation (MR) study aimed to further investigate their causal relationship. In this bidirectional MR study, the relationship between 73 proteins related to mitochondrial function and delirium, including delirium not induced by alcohol or other psychoactive substances (DEL) and delirium associated with alcohol withdrawal (AL-DEL). The random-effects inverse variance weighting (RE-IVW) method was used as the primary analytical method. Furthermore, multivariable MR (MVMR) analysis was performed to assess the impact of positive exposures and known risk factors for delirium. To ensure the reliability of our findings, heterogeneity and pleiotropy tests were conducted. The results of the RE-IVW method of MR analysis revealed that two proteins were positively associated with DEL (P < 0.05, odds ratio (OR) >1), whereas one protein was negatively associated with AL-DEL (P < 0.05, OR <1). In MVMR, ATP synthase subunit beta (ATP5F1B) was positively associated with DEL (P < 0.05, OR >1). Moreover, reverse MR analysis demonstrated that DEL was positively associated with three proteins (P < 0.05, OR >1) and negatively associated with two proteins (P < 0.05, OR <1). Finally, none of these associations displayed heterogeneity and horizontal pleiotropy (P > 0.05) or reverse causality. This bidirectional, multivariable two-sample MR analysis identified a causal relationship between eight proteins related to mitochondrial function and delirium. These findings offer novel insights that could potentially influence early diagnosis, expand our understanding of the underlying mechanisms, and inform treatment strategies for delirium. Nevertheless, given the possibility of bias, these results should be interpreted with caution.

First Report of Leaf Blight on Paris polyphylla Caused by Ceratobasidium ramicola in China.

Paris polyphylla var. yunnanensisis a perennial herb with significant medicinal properties in anticancer, anti-inflammatory, antibacterial immunomodulatory and antispasmodic activities (Duan et al. 2018). In April 2022, leaf blight disease emerged in Xiangtan City (Hunan), affecting P. polyphylla plantings over an area of 3×104 m2 (27.904°N, 112.918°E). The disease incidence reached an average of 22% of the plants in the field, with infected plants initially displaying water-soaked chlorosis, followed by dry yellow shrinkage that gradually spread from the leaf tips to the entire plant. To identify the causal agent, 20 leaf lesions (4 mm2) collected from 20 plants were surface sterilized with 75% ethanol for 10 s, 5% NaOCl for 30 s, rinsed in sterile distilled water three times, transferred to potato dextrose agar (PDA) plates with lactic acid (0.125%) , and incubated at 28 °C in the dark. Four isolates (PP21 to PP24) with similar morphologies were obtained and purified by the hyphal-tip method. Colonies on PDA initially appeared white with cottony mycelium, later turning light-yellow on the underside. Septate hyphae were branched at right angles with a small constriction at the departure of the branch point and measured 4.16 to 7.93 µm in diameter. Binucleate cells within the septate hyphae were visualized using Giemsa staining (Servicebio, China). For molecular identification, the rDNA internal transcribed spacer (ITS), the second largest subunit of nuclear DNA-directed RNA polymerase II (RPB2), and ATP synthase subunit 6 (ATP6) were amplified from genomic DNA of the isolates extracted by Fungus Genomic DNA Extraction Kit (Bioflux, China) using primers ITS1/ITS4 (White et al. 1990), bRPB2-6F/bRPB2-7.1R (Matheny 2005; Reeb et al. 2004) and ATP61/ATP62 (Kretzer and Bruns 1999), respectively. The ITS, RBP2 and ATP6 of the four isolates were sequenced and deposited in GenBank. BLASTn search of sequenced ITS (PQ187050, PQ187051, PP728052, PQ187052), RBP2 (PQ202833, PQ202834, PP735921, PQ202835) and ATP6 (PQ202837, PQ202838, PP735922, PQ202839) revealed a >99% identity with the type strain of Ceratobasidium ramicola CBS133.82 (NR138368, DQ301708, and DQ301577). For phylogenetic analysis, concatenated sequences of ITS, RBP2, and ATP6 were employed using the maximum-likelihood method in MEGA-X. Based on the morphological and molecular analyses, the isolates were classified into the C. ramicola clade (Bandoni 1979; Samuels et al. 2012; Jeong et al. 2023). To test the pathogenicity of the isolate PP23, mycelial plugs (5 mm in diameter) were placed directly on 15 healthy leaves from 15 three-year-old plants after puncturing with sterile needles. Sterile PDA plugs served as controls. All plants were kept in a greenhouse with conditions of 25°C, 80% relative humidity and a photoperiod of 12 h. After 5 days, all infected plants developed leaf blight symptoms similar to those described above, whereas the control plants remained asymptomatic. The pathogenicity test was conducted three times. C. ramicolawas re-isolated from all symptomatic leaves and confirmed to be identical to the original isolate based on morphology and nucleotide sequences of ITS, RBP2, and ATP6. While C. ramicola is commonly reported to be isolated from diseased cacao, its pathogenicity to cacao remains unknown (Samuels et al. 2012). To our knowledge, this is the first report of C. ramicola causing leaf blight on P. polyphylla, a medicinal herb with significant economic importance in China.

Structure of yeast RAVE bound to a partial V1 complex

Vacuolar-type ATPases (V-ATPases) are membrane-embedded proton pumps that acidify intracellular compartments in almost all eukaryotic cells. Homologous with ATP synthases, these multisubunit enzymes consist of a soluble catalytic V1 subcomplex and a membrane-embedded proton-translocating VO subcomplex. The V1 and VO subcomplexes can undergo reversible dissociation to regulate proton pumping, with reassociation of V1 and VO requiring the protein complex known as RAVE (regulator of the ATPase of vacuoles and endosomes). In the yeast Saccharomyces cerevisiae, RAVE consists of subunits Rav1p, Rav2p, and Skp1p. We used electron cryomicroscopy (cryo-EM) to determine a structure of yeast RAVE bound to V1. In the structure, RAVE is an L-shaped complex with Rav2p pointing toward the membrane and Skp1p distant from both the membrane and V1. Only Rav1p interacts with V1, binding to a region of subunit A not found in the corresponding ATP synthase subunit. When bound to RAVE, V1 is in a rotational state suitable for binding the free VO complex, but in the structure, it is partially disrupted, missing five of its 16 subunits. Other than these missing subunits and the conformation of the inhibitory subunit H, the V1 complex with RAVE appears poised for reassembly with VO.

Effect of Chloroplast ATP Synthase on Reactive Oxygen Species Metabolism in Cotton.

Abnormal programmed cell death in the tapetum is induced by reactive oxygen species (ROS), which are the main factors leading to cytoplasmic male sterility (CMS). These abnormalities are caused by genetic interactions between nuclear and cytoplasmic genes. To explore the role of chloroplast genes in ROS metabolism, next-generation and single-molecule real-time sequencing of the chloroplast genome were performed in the cotton CMS line Jin A (Jin A-CMS). Our results showed that the chloroplast genome is 160,042 bp in length and consists of 131 genes, including 112 functional genes. An analysis of the functional annotation and sequence comparison with the Gossypium hirsutum chloroplast genome as a reference revealed that 29 genes in Jin A-CMS have single-nucleotide polymorphisms, including subunits of ATP synthase, NAD(P)H-quinone redox reductase, and photosystem complexes. Compared to the Jin B maintainer, the anthers of Jin A-CMS at the microspore abortion stage have significantly lower expression of atpB, atpE, and atpF. The relative expression of these genes is significantly higher in the three-line F1 hybrids compared to Jin A-CMS. The ROS levels in the leaves increased in response to the silencing of atpE and atpF in cotton plants. In summary, the results of our study show that the ATP synthase subunit genes atpE and atpF are closely linked with ROS metabolism. These results provide basic information for the functional analysis of ATP synthase in cotton.

Novel Interacting Partners of MGP-40, a Chitinase-Like Protein in Buffalo Mammary Epithelial Cells.

Mammary Gland Protein-40 (MGP-40), also known as chitinase-3-like protein 1 (CHI3L1), is involved in critical biological processes such as inflammation, tissue remodeling, and cell proliferation, especially during the involution phase of the mammary gland. This study aimed to explore the molecular mechanisms of MGP-40 by identifying its novel interacting partners in buffalo mammary epithelial cells (BuMECs). Stable overexpression of MGP-40 in BuMECs was achieved through transfection with the pCIneo-MGP-40 vector, followed by G418 selection and confirmation by Western blot analysis. To identify interacting proteins, Co-immunoprecipitation (Co-IP) of BuMEC lysate using an anti-YKL-40 antibody was performed, and the eluted proteins were analyzed using SDS-PAGE and mass spectrometry (MALDI-TOF/TOF). The analysis revealed several interacting proteins, including synaptotagmin-like 3, Ras-related Rab19, RIB34A-like protein with coiled coils, and ATP synthase subunit g. These interacting partners suggest that MGP-40 is involved in crucial cellular processes like vesicle trafficking, cytoskeletal organization, and energy metabolism, extending its known functions in inflammation and tissue remodeling. Notably, the interactions with synaptotagmin-like 3 and Rab proteins emphasize MGP-40's potential role in vesicular transport, essential for milk production in mammary epithelial cells, while the association with ATP synthase subunit g links MGP-40 to energy regulation during lactation. These findings provide preliminary insights into the potential roles of MGP-40 in mammary gland physiology, particularly in cellular processes such as vesicle trafficking and energy metabolism. Further studies, including in vivo validation, are essential to confirm these interactions and clarify their relevance to mammary gland function and pathology.

Transcriptome and interactome-based analyses to unravel crucial proteins and pathways involved in Acinetobacter baumannii pathogenesis.

The present study employed an integrated transcriptome and interactome-based analyses to identify key proteins and pathways associated with Acinetobacter baumannii infection towards the development of novel therapeutics against this pathogen. Transcriptome analysis of A.baumannii strains (ATCC 17978 and AbH12O-A2) identified 253 and 619 differentially expressed genes (DEGs), respectively. These genes were involved in essential molecular functions, including DNA binding, metal ion binding, and oxidoreductase activity. The centrality and module analyses of these identified DEGs had shortlisted 27 and 41 hub proteins, which were central to the ATCC 17978 and AbH12O-A2 networks, and essential for bacterial survival. Significantly, three proteins (SecA, glutathione synthase, and aromatic-amino-acid transaminase) from the ATCC 17978 strain and seven proteins (ATP synthase subunit alpha, translation initiation factor IF-2, SecY, elongation factors G, Tu, and Ts, and tRNA guanine-N1-methyltransferase) from the AbH12O-A2 strain showed interactions with human proteins, identified through host-pathogen interaction (HPI) analysis of hub proteins (referred as hub-HPI proteins). These proteins were observed to participate in vital pathways, including glutathione metabolism, secondary metabolite biosynthesis and quorum sensing. Targeting these hub-HPI proteins through novel therapeutic strategies holds the potential to disrupt the critical bacterial pathways, thereby controlling A. baumannii infections. Furthermore, their localization analysis indicated that nine proteins were cytoplasmic and one was membrane protein. Among them, six were druggable and four were novel proteins. Overall, this comprehensive study provides valuable insights into the crucial proteins and pathways involved during A. baumannii infection, and offers potential therapeutic targets for designing novel antimicrobial agents to tackle the pathogen.

Levilactobacillus brevis 47f: Bioadaptation to Low Doses of Xenobiotics in Aquaculture.

Agricultural and industrial activities are increasing pollution of water bodies with low doses of xenobiotics that have detrimental effects on aquaculture. The aim of this work was to determine the possibility of using Levilactobacillus brevis 47f culture in fish aquaculture under the influence of low doses of xenobiotics as an adaptogen. An increase in the survival of Danio rerio individuals exposed to the xenobiotic bisphenol A solution and fed with the L. brevis 47f was shown compared to control groups and, at the same time, the cytokine profile in the intestinal tissues of Danio rerio was also investigated. Analysis of differential gene expression of the L. brevis 47f grown under the action of high concentrations of bisphenol A showed changes in mRNA levels of a number of genes, including genes of various transport proteins, genes involved in fatty acid synthesis, genes of transcriptional regulators, genes of the arabinose operon, and the oppA gene. The identification of L. brevis 47f proteins from polyacrylamide gel by mass spectrometry revealed L-arabinose isomerase, Clp chaperone subunit, ATP synthase subunits, pentose phosphate pathway and glycolysis enzyme proteins, which are likely part of the L. brevis 47f strain's anti-stress response, but probably do not affect its adaptogenic activity toward Danio rerio.

Cucumber Green Mottle Mosaic Virus Coat Protein Hijacks Mitochondrial ATPδ to Promote Viral Infection.

The production and scavenging of reactive oxygen species (ROS) are critical for plants to adapt to biotic and abiotic stresses. In this study, we investigated the interaction between the coat protein (CP) of cucumber green mottle mosaic virus (CGMMV) and ATP synthase subunit δ (ATPδ) in mitochondria. Silencing of ATPδ by tobacco rattle virus-based virus-induced gene silencing impeded CGMMV accumulation in Nicotiana benthamiana leaves. Both the overexpression of ATPδ in transgenic plants and transient expression promoted CGMMV infection. Nitro blue tetrazolium (NBT) and 3,3'-diaminobenzidine (DAB) staining revealed that ATPδ inhibited O2 - production but not H2O2 production. The treatment of CGMMV-infected leaves with the ROS inhibitor diphenylene iodonium (DPI) induced a ROS burst that inhibited CGMMV infection. Reverse transcription-quantitative PCR and superoxide dismutase (SOD) activity assays showed that ATPδ, CGMMV infection, and CP expression specifically induced NbFeSOD3/4 expression and SOD activity, and silencing NbFeSOD3/4 inhibited CGMMV infection. We speculate that CGMMV CP interacts with ATPδ and hijacks it, thereby enhancing O2 - quenching by upregulating NbFeSOD expression and, in turn, SOD activity.

Reproductive effort affects cellular response in the mantle of Nodipecten subnodosus scallops exposed to acute hyperthermia

In marine ecosystems, temperature regulates the energy metabolism of animals. In the last decades, the temperature increase was related to mass mortality events of marine ectotherms, particularly during high-energy investment for reproduction. In scallops, the mantle has been poorly investigated while this tissue covers more than 40 % of the body mass, contributing to the perception of surrounding environmental stimuli. Our aim was to assess the cellular and molecular responses linked to energy metabolism in the mantle of adult N. subnodosus facing acute hyperthermia during reproductive effort. Scallops collected in spring (late gametogenesis) and summer (ripe gonads) were exposed to a control temperature (22 °C) or acute hyperthermia (30 °C) for 24 h. In spring, increased arginine kinase (AK) activity together with increased pyruvate kinase/citrate synthase ratio (PK/CS) suggested an enhanced carbohydrate, pyruvate, and arginine metabolism to maintain the adenylate energy charge (AEC) in the mantle of scallops coping with acute thermal increase. In summer, animals decreased their AEC (5 %) and arginine phosphate pool (40 %) and increased their anaerobic metabolism as shown by enhanced activities of lactate dehydrogenase (LDH) and octopine dehydrogenase (ODH), respectively. The abundance of twenty proteins involved in energy metabolism (isocitrate dehydrogenase, ATP synthase subunit β), protein protection (cognate heat shock protein 70), and cytoskeleton (actins and tubulins) were affected only by season. These results underlie the role of the mantle of N. subnodosus in the seasonal responses of this tissue to thermal fluctuations during reproductive effort with possible implications for the physiological performance of scallops under heat waves in wild or harvest conditions.

Graphene quantum dots disrupt the mitochondrial potential of Trypanosoma brucei by interacting with the p18 subunit of ATP synthase F1 after endocytosis via the VSG recycling pathway

HypothesisTrypanosomiasis is one of the main threats to human and animal health in African countries. Trypanosoma brucei can evade the host immune recognition by rapidly altering its variant surface glycoprotein (VSG). The ATP synthase F1 subunit of the parasite exhibits extremely low similarity to that of its mammalian hosts, hypothetically making it an ideal target for the development of novel therapeutics. ExperimentsGraphene quantum dots (GQDs) were synthesized, and their adhesion to T. brucei surface and internalization was observed microscopically. The activity of ATP synthase and mitochondrial membrane potential of T. brucei were measured after exposure to GQDs. Proteomics, biolayer interferometry, and molecular dynamic simulations were utilized to evaluate the interaction between GQDs with the target proteins. FindingsGQDs specifically adhered to the VSG of T. brucei and were conveyed inside the parasite via the VSG internalization pathway. The GQDs promoted intracellular ROS production, interacted with, and inhibited the activity of the p18 subunit of ATP synthase, disrupted parasite mitochondrial membrane potential. Additionally, the GQDs caused a decrease in aminoacyl – tRNA biosynthesis, and upregulated RNA and protein degradation pathways. The findings of this study offer a novel avenue for the target-oriented discovery of anti-trypanosome drugs.

Ginsenoside Rg_1 modulates ATP5A1 deacetylation via SIRT3 to attenuate hypoxia/reoxygenation injury in HL-1 cardiomyocytes

This study investigated the mechanism by which ginsenoside Rg_(1 )attenuates hypoxia/reoxygenation(H/R) injury in HL-1 cardiomyocytes by inhibiting the acetylation of ATP synthase subunit alpha(ATP5A1) through silent information regulator 3(SIRT3). In this study, an H/R injury model was constructed by hypoxia for 6 h and reoxygenation for 2 h in HL-1 cardiomyocytes. First, the optimal effective concentration of ginsenoside Rg_1 was determined using a cell viability assay kit. Then, lactate dehydrogenase(LDH) leakage was measured using a microplate method to evaluate the protective effect of ginsenoside Rg_1 against H/R injury in HL-1 cardiomyocytes. Western blot was further used to detect SIRT3 expression and the acetylation level of mitochondrial proteins in cardiomyocytes. ATP content was measured using a luciferase assay. Immunoprecipitation was used to detect the acetylation level of ATP5A1. The oxygen consumption rate(OCR) was measured using the Seahorse XFp analyzer. Flow cytometry was used to assess cell apoptosis to explore the specific mechanism. The results showed that compared with the control group, the model group had a significant increase in LDH leakage, a decrease in SIRT3 expression, a significant increase in the acetylation levels of mitochondrial proteins and ATP5A1, a decrease in OCR and ATP content, and an increase in cell apoptosis rate. Compared with the model group, the ginsenoside Rg_1 group showed a significant decrease in LDH leakage, an increase in SIRT3 expression, a significant decrease in the acetylation levels of mitochondrial proteins and ATP5A1, an increase in OCR and ATP content, and a decrease in cell apoptosis rate. Compared with the ginsenoside Rg_1 group, the ginsenoside Rg_1 + si SIRT3 group showed a significant increase in LDH leakage, a decrease in SIRT3 expression, a significant increase in the acetylation levels of mitochondrial proteins and ATP5A1, a decrease in OCR and ATP content, and an increase in cell apoptosis rate. In conclusion, ginsenoside Rg_1 alleviates H/R injury in HL-1 cells by improving mitochondrial respiratory function through SIRT3-mediated inhibition of ATP5A1 acetylation.