{"gene":"NDUFS1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2001,"finding":"Point mutations and large-scale deletions in the NDUFS1 gene (encoding the 75-kDa Fe-S subunit of mitochondrial complex I) cause isolated complex I deficiency in humans, establishing NDUFS1 as a nuclear-encoded structural subunit required for complex I activity.","method":"Denaturing HPLC and direct cDNA sequencing of NDUFS1 in patient fibroblasts; biochemical respiratory chain enzyme assays","journal":"American journal of human genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct sequencing plus biochemical activity assays in patient-derived cells, single study but multiple mutations and patients","pmids":["11349233"],"is_preprint":false},{"year":2006,"finding":"A C1564A missense mutation (Q522K) in NDUFS1 reduces the level of mature complex I, markedly inhibits NADH-ubiquinone oxidoreductase activity, causes accumulation of mitochondrial H2O2 and superoxide, decreases mitochondrial potential, and leads to glutathione depletion; ROS increase was not observed in the NDUFS4 mutant, demonstrating a specific role for the Fe-S NDUFS1 subunit in electron transfer and ROS generation.","method":"Biochemical assays in patient fibroblasts: complex I activity measurement, ROS detection (H2O2, O2•−), mitochondrial membrane potential measurement, glutathione quantification, glutathione peroxidase activity assay; dibutyryl-cAMP rescue experiment","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal biochemical methods in patient cells with mechanistic rescue experiment, single lab","pmids":["16478720"],"is_preprint":false},{"year":2010,"finding":"Novel NDUFS1 mutations (including a premature stop, amino acid substitutions, and a single-amino-acid deletion) cause decreased complex I amount and activity and a disturbed complex I assembly pattern in patient fibroblasts, establishing NDUFS1 as required for proper assembly and stability of mitochondrial complex I.","method":"NDUFS1 gene sequencing in patient fibroblasts; complex I activity assay; Blue Native PAGE assembly analysis","journal":"Molecular genetics and metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical activity and native PAGE assembly assays across three independent patients, single lab","pmids":["20382551"],"is_preprint":false},{"year":2011,"finding":"A homozygous p.Thr595Ala mutation in NDUFS1 causes severe reduction of complex I enzyme activity in muscle and complex I dysfunction in a Neurospora crassa insertional mutagenesis model and in patient fibroblasts grown in galactose, providing cross-species genetic evidence that NDUFS1 is functionally conserved and essential for complex I activity.","method":"Muscle biopsy complex I enzyme activity assay; Neurospora crassa insertional mutagenesis model; patient fibroblasts grown in galactose (stress condition to unmask OXPHOS defect)","journal":"Neurogenetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — orthologous model organism mutagenesis plus patient biochemistry, single study","pmids":["21203893"],"is_preprint":false},{"year":2013,"finding":"Caspase-3 cleaves the p75 NDUFS1 subunit of respiratory complex I downstream of MOMP during TNFα+cycloheximide-induced apoptosis; this cleavage drives ROS formation, which then triggers lysosomal membrane permeability (LMP) and cathepsin release, amplifying apoptosis. A caspase-non-cleavable p75 mutant prevented LMP, confirming the NDUFS1 cleavage event as mechanistically causal.","method":"Genetic epistasis with Bax/Bak, Apaf-1, caspase-9, caspase-3/7 double-knockout cells; caspase-non-cleavable NDUFS1 mutant expression; MitoQ antioxidant rescue; LMP and cathepsin release assays; ROS measurement","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — mutagenesis of cleavage site combined with genetic KO epistasis and pharmacological rescue across multiple orthogonal readouts","pmids":["23788428"],"is_preprint":false},{"year":2019,"finding":"MDM2 directly binds NDUFS1 via its amino-terminal region (aa 1–101), sequesters it in the cytoplasm, prevents its mitochondrial localization, and thereby destabilizes complex I and respiratory supercomplexes, leading to decreased mitochondrial respiration, oxidative stress, and commitment to the mitochondrial apoptosis pathway in a p53-independent manner.","method":"Complementary biochemical (Co-IP, pull-down), organellar fractionation, and cellular approaches; MDM2 amino-terminal truncation mapping; supercomplex analysis by BN-PAGE; Drosophila and murine transgenic Mdm2 models; oxygen consumption rate assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct binding demonstrated by pull-down with domain mapping, replicated in two animal models, multiple orthogonal functional readouts","pmids":["30879903"],"is_preprint":false},{"year":2019,"finding":"Biallelic NDUFS1 mutations decrease the stability of the entire N-module of complex I and disrupt electron transfer between two iron-sulfur clusters within NDUFS1, causing metabolic reprogramming including TCA cycle inhibitory feedback and elevated reactive oxygen species stress.","method":"Proteome and metabolome profiling of patient-derived cells; structural inference from iron-sulfur cluster positions; comparison with a second CI gene mutation","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteomics plus metabolomics in patient cells with mechanistic interpretation from structural context, single lab","pmids":["31557978"],"is_preprint":false},{"year":2020,"finding":"AKAP1 interacts with NDUFS1 (identified by immunoprecipitation and mass spectrometry) and is required for translocation of NDUFS1 from the cytosol to mitochondria; AKAP1 deficiency prevents this translocation, inhibits complex I activity, reduces ATP production, increases mitochondrial ROS, and exacerbates cardiomyocyte apoptosis. Restoration of AKAP1 rescues mitochondrial NDUFS1 localization and cardiac function.","method":"Co-immunoprecipitation and LC-MS/MS; Akap1-KO mice with STZ-induced diabetes; AAV9-Akap1 cardiac overexpression rescue; echocardiography; complex I activity assay; ROS measurement; subcellular fractionation","journal":"Diabetologia","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP/MS plus in vivo KO and AAV rescue with multiple functional readouts establishing the AKAP1–NDUFS1 mitochondrial import mechanism","pmids":["32072193"],"is_preprint":false},{"year":2021,"finding":"NDUFS1 knockdown in cardiomyocytes decreases mitochondrial DNA content, mitochondrial membrane potential, and mitochondrial mass while increasing mitochondrial ROS production; Ndufs1 overexpression reverses Ang II-induced cardiomyocyte hypertrophy phenotypes, establishing a direct role for NDUFS1 in maintaining mitochondrial membrane potential in cardiomyocytes.","method":"siRNA knockdown and overexpression of Ndufs1 in rat cardiomyocytes; Ang II hypertrophy model; MMP measurement (JC-1), mtDNA content, mitochondrial mass, and ROS assays","journal":"Oxidative medicine and cellular longevity","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss- and gain-of-function with multiple mitochondrial phenotype readouts, single lab","pmids":["33763166"],"is_preprint":false},{"year":2022,"finding":"PHB2 directly interacts with NDUFS1 (identified by Co-IP and mass spectrometry) and co-localizes with it in mitochondria; this interaction facilitates NDUFS1 binding to NDUFV1, stabilizes complex I, and enhances complex I activity, thereby elevating oxidative phosphorylation levels in colorectal cancer cells.","method":"Co-immunoprecipitation and mass spectrometry; confocal co-localization; complex I activity assay after PHB2 knockdown or overexpression; PHB2 KD combined with PHB2 OE rescue","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP/MS plus co-localization and functional complex I activity assay, single lab","pmids":["36658121"],"is_preprint":false},{"year":2022,"finding":"Glutathionylation of NDUFS1 within complex I (induced by disulfiram) increases mitochondrial superoxide/H2O2 production during reverse electron transfer from the ubiquinone pool via substrates glycerol-3-phosphate and proline; deglutathionylation of NDUFS1 by reducing agents restores normal complex I activity and decreases ROS production.","method":"Immunocapture of complex I from liver mitochondria; disulfiram-induced glutathionylation; site-specific inhibitors for complex I, III, GPD, PRODH; ROS measurement; reducing-agent reversal of glutathionylation","journal":"Antioxidants (Basel, Switzerland)","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — in vitro mitochondrial assay with immunocapture, pharmacological dissection of ROS sites, and reversible modification, single lab","pmids":["36290766"],"is_preprint":false},{"year":2022,"finding":"Mutations at NDUFS1-corresponding positions in the homologous E. coli nuoG subunit reduce NADH oxidase activity and disrupt complex I assembly (assessed by co-immunoprecipitation and time-delayed expression assays), and many map to subunit interfaces; compound heterozygote modeling identified which mutation in a pair is more deleterious.","method":"Site-directed mutagenesis in E. coli nuoG (NDUFS1 ortholog); membrane vesicle NADH oxidase activity assay; co-immunoprecipitation assembly assay; time-delayed expression assay; alanine substitution series","journal":"Mitochondrion","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution in bacterial model with mutagenesis and assembly assays, single lab, ortholog model","pmids":["36462614"],"is_preprint":false},{"year":2023,"finding":"Reduction of NDUFS1 in gastric cancer cells activates the mitochondrial ROS–HIF1α signaling pathway, upregulating FBLN5 (a transcriptional target of HIF1α), thereby promoting cancer cell proliferation, migration, and invasion; NDUFS1 overexpression suppresses this pathway and inhibits tumor growth in vivo.","method":"Confocal microscopy for NDUFS1 subcellular localization and mROS measurement; CCK-8, colony formation, transwell assays; mouse xenograft model; western blot and IHC for pathway components","journal":"British journal of cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss- and gain-of-function with in vivo xenograft, pathway component measurement, and localization imaging, single lab","pmids":["37644092"],"is_preprint":false},{"year":2024,"finding":"Agrimol B causes caspase-3-mediated degradation of NDUFS1 protein, leading to mitochondrial ROS accumulation, autophagosome-lysosome fusion blockade (autophagy arrest), and HCC cell growth inhibition; NDUFS1 overexpression partially restores mitochondrial ROS levels and reverses autophagy arrest induced by agrimol B.","method":"NDUFS1 overexpression rescue experiment; caspase-3 activity assay; mROS measurement; autophagosome accumulation assay; in vitro and PDX in vivo models","journal":"Free radical biology & medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic rescue with NDUFS1 OE and caspase-3 pathway dissection, in vivo PDX validation, single lab","pmids":["38697493"],"is_preprint":false},{"year":2024,"finding":"RNF43 (an E3 ubiquitin ligase) directly interacts with NDUFS1 and promotes its ubiquitination and proteasomal degradation, reducing oxidative phosphorylation activity; NDUFS1 is thus a downstream target of RNF43 in endometrial stromal cells.","method":"Co-immunoprecipitation demonstrating RNF43–NDUFS1 interaction; ubiquitination assay; NDUFS1 knockdown phenocopy of RNF43 overexpression; OXPHOS activity measurement","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP establishing interaction plus ubiquitination assay and functional rescue, single lab","pmids":["38988031"],"is_preprint":false},{"year":2025,"finding":"Berberine directly binds and activates SIRT3, which deacetylates NDUFS1 (the catalytic subunit in the N-module of complex I), causing dissociation of complex I from the mitochondrial membrane; this selectively and reversibly reduces complex I abundance and OXPHOS activity in hepatocytes, improving glucose and lipid metabolism.","method":"In vivo oral administration followed by mitochondrial isolation; SIRT3 activation assay; acetylation state of NDUFS1 measured by IP; complex I dissociation by BN-PAGE; oxygen consumption rate; glucose/lipid metabolic readouts","journal":"Science China. Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding of berberine to SIRT3 linked to NDUFS1 deacetylation and complex I dissociation with multiple orthogonal readouts, single lab","pmids":["40493314"],"is_preprint":false},{"year":2025,"finding":"PCBP2 binds NDUFS1 mRNA (verified by RNA-immunoprecipitation and RNA-protein pull-down), stabilizes it, and promotes NDUFS1 protein expression; increased NDUFS1 in turn activates NRF2 nuclear translocation, inhibiting cardiomyocyte ferroptosis during myocardial infarction.","method":"RNA-immunoprecipitation (RIP); RNA-protein pull-down; NDUFS1 overexpression and PCBP2 overexpression experiments; NRF2 nuclear translocation assay; ferroptosis markers; in vivo MI mouse model with LV-PCBP2/LV-NDUFS1","journal":"Molecular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RIP and pull-down establishing RNA-protein interaction, plus in vivo rescue, single lab","pmids":["40784311"],"is_preprint":false},{"year":2025,"finding":"Caspase-3 cleaves NDUFS1 at residue D255; mutation D255A abolishes this cleavage and attenuates ROS accumulation and mitochondrial dysfunction induced by trichothecene mycotoxins, confirming that caspase-3-mediated NDUFS1 cleavage disrupts electron transport and amplifies mitochondrial ROS in a positive feedback loop with ERO1α-mediated ER oxidative stress.","method":"In vivo and in vitro mycotoxin exposure models; caspase-3 inhibition and siRNA knockdown; NDUFS1 D255A cleavage-site mutant expression; ROS measurement; mitochondrial function assays","journal":"Free radical biology & medicine","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct site mutagenesis (D255A) preventing caspase-3 cleavage with functional rescue in both in vitro and in vivo models, single lab but two orthogonal approaches","pmids":["41422996"],"is_preprint":false},{"year":2025,"finding":"NDUFS1 deficiency in alveolar epithelial cells reduces complex I activity, impairs NAD+ production, and increases ROS, which in turn decreases ENaCα expression and impairs alveolar fluid clearance; supplementing NAD+ via Olaparib restores ENaCα levels and alleviates acute lung injury phenotypes caused by NDUFS1 deficiency.","method":"NDUFS1 knockdown in alveolar epithelial cells; complex I activity assay; NAD+ measurement; ROS assay; ENaCα expression; Olaparib-mediated NAD+ supplementation rescue; ALI mouse models","journal":"International journal of medical sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KD plus pharmacological rescue establishing NAD+/ENaCα as mechanistic mediators downstream of NDUFS1, single lab","pmids":["40860777"],"is_preprint":false},{"year":2025,"finding":"Naringin facilitates translocation of NDUFS1 from the cytosol to mitochondria in cardiac microvascular endothelial cells during hypoxia-reoxygenation injury; this mitochondrial import of NDUFS1 restores mitochondrial function, reduces ROS, and suppresses ferroptosis via the IRF3/SLC7A11/GPX4 axis.","method":"Immunofluorescence and subcellular fractionation to track NDUFS1 localization; proteomic analysis; molecular docking and molecular dynamics; ferroptosis marker assays; in vivo MI/RI rat model","journal":"The Journal of nutritional biochemistry","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization by immunofluorescence with functional correlation but no direct interaction mapping; single lab, single study","pmids":["40617306"],"is_preprint":false},{"year":2026,"finding":"GL-V9 binds the MDM2 amino-terminal domain (aa 1–101) and acts as a molecular glue that facilitates MDM2–NDUFS1 interaction in the cytoplasm, preventing NDUFS1 mitochondrial localization, inhibiting complex I formation, disrupting mitochondrial homeostasis, and activating the OMA1-DELE1 integrated stress response to induce apoptosis, in a p53-independent manner.","method":"GST pull-down assay; cellular thermal shift assay (CETSA); surface plasmon resonance (SPR); immunofluorescence for NDUFS1 mitochondrial localization; MDM2 amino acid mutation mapping; mitochondrial membrane potential, superoxide, ATP, OCR assays; OMA1-DELE1 pathway readout","journal":"Journal of advanced research","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — direct binding confirmed by SPR, CETSA, GST pull-down, and amino acid mutagenesis; NDUFS1 localization and complex I function validated by multiple orthogonal assays; single lab","pmids":["41951044"],"is_preprint":false},{"year":2026,"finding":"NDUFS1-mediated complex I activity maintains pancreatic cancer stem cell stemness and tumorigenicity; mechanistically, CD147 promotes pSTAT3Tyr705-mediated NDUFS1 transcription, and NDUFS1 initiates SIRT1-DNMT1 metaboloepigenetic signaling that reduces PAX2 promoter methylation, increasing PAX2 expression to sustain stemness.","method":"ALDH+ CSC sorting and tumorsphere assay; complex I activity assays; NDUFS1 KD/OE; ChIP for PAX2 promoter methylation; DNMT1/SIRT1 pathway analysis; CD147 overexpression; in vivo tumorigenicity assays","journal":"MedComm","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal functional assays linking NDUFS1 to epigenetic regulation of stemness, single lab","pmids":["41930324"],"is_preprint":false},{"year":2026,"finding":"NDUFS1 K170 lactylation (induced during ischemia-reperfusion) impairs the cardioprotective effects of PG; overexpression of NDUFS1 K170 lactylation diminished PG-mediated improvement of MIRI, establishing a specific lysine lactylation site on NDUFS1 as a post-translational modification that modulates mitochondrial function and ferroptosis in myocardial injury.","method":"Multi-omics (metabolomics, proteomics); overexpression of NDUFS1 K170 lactylation mutant; PDK4 overexpression; in vivo rat MI/RI model; GPX4, ACSL4, PDK4 pathway readouts","journal":"Redox biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — site-specific lactylation mutant with in vivo functional readout, single lab, single study","pmids":["42134300"],"is_preprint":false},{"year":2025,"finding":"ATF3, upregulated downstream of the ER stress PERK-eIF2α-ATF4 pathway by sorafenib, negatively regulates NDUFS1 expression; siRNA silencing of ATF3 partially restores mitochondrial function impaired by sorafenib, defining an ATF3→NDUFS1 regulatory axis in sorafenib-induced cardiotoxicity.","method":"Transcriptomic and proteomic profiling; ATF3 siRNA knockdown; Western blot validation; mitochondrial function assays; ER stress inhibitor (GSK2606414) rescue; H9C2 cell model and in vivo rat model","journal":"Frontiers in pharmacology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — ATF3 KD with functional rescue but mechanistic link to NDUFS1 relies primarily on proteomic screening without direct ATF3–NDUFS1 promoter or binding assay, single lab","pmids":["40880646"],"is_preprint":false}],"current_model":"NDUFS1 (CI-75k) encodes the 75-kDa iron-sulfur subunit that forms the core of the N-module of mitochondrial respiratory chain complex I, where it coordinates iron-sulfur clusters essential for electron transfer from NADH to ubiquinone; it must be imported from the cytosol into mitochondria (a step facilitated by AKAP1 and antagonized by MDM2 sequestration), assembles into complex I through subunit interfaces (where pathogenic mutations disrupt assembly and activity), and its activity is regulated post-translationally by glutathionylation (increasing ROS via reverse electron transfer), SIRT3-dependent deacetylation (leading to complex I dissociation), and lactylation at K170; caspase-3 cleaves NDUFS1 during apoptosis to amplify mitochondrial ROS and lysosomal membrane permeability, while binding partners including PHB2, CD147-STAT3, and RNF43 modulate its stability and activity to control oxidative phosphorylation, ROS production, and downstream signaling in diverse cellular contexts."},"narrative":{"mechanistic_narrative":"NDUFS1 encodes the 75-kDa iron-sulfur subunit that forms the catalytic core of the N-module of mitochondrial respiratory complex I, where it coordinates iron-sulfur clusters that relay electrons from NADH toward ubiquinone [PMID:11349233, PMID:31557978]. Pathogenic point mutations, deletions, and single-residue substitutions reduce complex I amount and NADH-ubiquinone oxidoreductase activity and disrupt assembly and stability of the entire N-module, causing isolated complex I deficiency in humans; many disease residues map to subunit interfaces and to the electron-transfer path between two internal Fe-S clusters, and loss of activity drives accumulation of mitochondrial superoxide and H2O2, membrane-potential collapse, and glutathione depletion [PMID:11349233, PMID:16478720, PMID:20382551, PMID:31557978, PMID:36462614]. Because NDUFS1 is nuclear-encoded, its function depends on import into mitochondria: AKAP1 binds NDUFS1 and is required for its cytosol-to-mitochondria translocation, whereas MDM2 directly binds the protein and sequesters it in the cytoplasm, destabilizing complex I and respiratory supercomplexes and committing cells to mitochondrial apoptosis in a p53-independent manner [PMID:30879903, PMID:32072193]. NDUFS1 abundance and activity are further tuned post-translationally and through protein and RNA partners: glutathionylation increases ROS via reverse electron transfer [PMID:36290766], SIRT3-dependent deacetylation dissociates complex I from the membrane [PMID:40493314], K170 lactylation modulates mitochondrial function in ischemic myocardium [PMID:42134300], RNF43 ubiquitinates it for proteasomal degradation [PMID:38988031], PHB2 stabilizes the NDUFS1-NDUFV1 interface to enhance OXPHOS [PMID:36658121], and PCBP2 binds and stabilizes NDUFS1 mRNA [PMID:40784311]. During apoptosis, caspase-3 cleaves NDUFS1 (at D255) to amplify mitochondrial ROS, which in turn triggers lysosomal membrane permeabilization and ER oxidative stress feedback [PMID:23788428, PMID:41422996]. Through these mechanisms NDUFS1 acts as a node controlling oxidative phosphorylation, ROS output, and downstream signaling — including HIF1α, NRF2, and ferroptosis-related pathways — in cardiac, epithelial, and cancer contexts [PMID:33763166, PMID:37644092, PMID:40784311, PMID:40860777, PMID:41930324].","teleology":[{"year":2001,"claim":"Established NDUFS1 as a nuclear-encoded structural subunit whose mutation directly causes a respiratory chain disease, answering whether the gene is required for complex I function in humans.","evidence":"dHPLC and cDNA sequencing of patient fibroblasts plus respiratory chain enzyme assays","pmids":["11349233"],"confidence":"Medium","gaps":["Did not resolve how individual mutations affect the electron-transfer path","No assembly-stage analysis"]},{"year":2006,"claim":"Showed the Fe-S NDUFS1 subunit specifically governs both electron transfer and ROS generation, distinguishing it from other complex I subunits whose loss does not raise ROS.","evidence":"Biochemical activity, ROS, membrane-potential and glutathione assays in Q522K patient fibroblasts with cAMP rescue","pmids":["16478720"],"confidence":"High","gaps":["Single mutation in one patient line","Mechanism of ROS amplification not at residue resolution"]},{"year":2010,"claim":"Demonstrated that NDUFS1 mutations disturb complex I assembly, not just steady-state activity, defining the subunit as required for complex assembly and stability.","evidence":"Gene sequencing, activity assays and Blue Native PAGE assembly analysis in three patients","pmids":["20382551"],"confidence":"Medium","gaps":["Assembly intermediates not structurally mapped","Single lab"]},{"year":2011,"claim":"Provided cross-species confirmation that NDUFS1 is functionally conserved and essential, addressing whether human findings reflect a core conserved role.","evidence":"Muscle enzymology, Neurospora crassa insertional mutant, and galactose-stressed patient fibroblasts","pmids":["21203893"],"confidence":"Medium","gaps":["Single homozygous mutation","No direct biochemical Fe-S cluster measurement"]},{"year":2013,"claim":"Defined a non-respiratory role: caspase-3 cleavage of NDUFS1 during apoptosis amplifies ROS and triggers lysosomal membrane permeabilization, placing the subunit in an apoptotic feed-forward circuit.","evidence":"Bax/Bak, Apaf-1, caspase-9, caspase-3/7 KO epistasis, caspase-non-cleavable mutant, and MitoQ rescue","pmids":["23788428"],"confidence":"High","gaps":["Exact cleavage residue not defined here","Generality across apoptotic stimuli untested"]},{"year":2019,"claim":"Identified MDM2 as a direct cytoplasmic sequestering partner that blocks NDUFS1 mitochondrial import to suppress respiration and promote apoptosis independently of p53.","evidence":"Co-IP/pull-down with N-terminal (aa 1-101) domain mapping, BN-PAGE supercomplex analysis, and Drosophila/mouse Mdm2 models","pmids":["30879903"],"confidence":"High","gaps":["How MDM2 selects NDUFS1 over other import cargo unclear","No structure of the MDM2-NDUFS1 interface"]},{"year":2019,"claim":"Linked NDUFS1 mutations to destabilization of the whole N-module and disrupted inter-cluster electron transfer, connecting genotype to metabolic reprogramming.","evidence":"Proteome/metabolome profiling of patient cells with structural inference from Fe-S cluster positions","pmids":["31557978"],"confidence":"Medium","gaps":["Electron-transfer disruption inferred, not directly measured","Single lab"]},{"year":2020,"claim":"Established AKAP1 as a factor required for NDUFS1 import into mitochondria, defining a positive regulator of its localization opposite to MDM2.","evidence":"Reciprocal Co-IP/LC-MS/MS, Akap1-KO diabetic mice, AAV9-Akap1 cardiac rescue and fractionation","pmids":["32072193"],"confidence":"High","gaps":["Whether AKAP1 acts at the import machinery directly is unresolved","Interaction interface not mapped"]},{"year":2021,"claim":"Showed NDUFS1 level controls cardiomyocyte mitochondrial mass, mtDNA content and membrane potential, linking the subunit to hypertrophy phenotypes.","evidence":"siRNA knockdown and overexpression in rat cardiomyocytes with Ang II hypertrophy model","pmids":["33763166"],"confidence":"Medium","gaps":["Mechanism connecting NDUFS1 to mtDNA content unclear","Single lab, rodent cells"]},{"year":2022,"claim":"Defined several regulators of NDUFS1: PHB2 stabilizes the NDUFS1-NDUFV1 interface to boost OXPHOS, glutathionylation drives reverse-electron-transfer ROS, and ortholog mutagenesis mapped activity/assembly to subunit interfaces.","evidence":"Co-IP/MS and complex I assays (PHB2), immunocaptured complex I with reversible glutathionylation, and E. coli nuoG site-directed mutagenesis","pmids":["36658121","36290766","36462614"],"confidence":"Medium","gaps":["Glutathionylation site on NDUFS1 not pinpointed","PHB2 effect shown in one cancer context"]},{"year":2023,"claim":"Connected NDUFS1 loss to mitochondrial ROS-HIF1α-FBLN5 signaling that promotes gastric cancer progression, framing NDUFS1 as a tumor suppressor in this context.","evidence":"Loss/gain-of-function with xenografts, localization imaging and pathway component blots/IHC","pmids":["37644092"],"confidence":"Medium","gaps":["Direct HIF1α induction kinetics not resolved","Single lab"]},{"year":2024,"claim":"Showed NDUFS1 degradation routes converge on ROS and downstream cell-fate outcomes: caspase-3-driven degradation arrests autophagy in liver cancer, and RNF43-mediated ubiquitination lowers OXPHOS in endometrial stroma.","evidence":"NDUFS1 overexpression rescue with caspase-3 dissection (agrimol B), and Co-IP plus ubiquitination assay (RNF43)","pmids":["38697493","38988031"],"confidence":"Medium","gaps":["RNF43 ubiquitination site on NDUFS1 not mapped","Pathway specificity across tissues unclear"]},{"year":2025,"claim":"Expanded the post-translational and transcriptional control of NDUFS1: SIRT3 deacetylation dissociates complex I from the membrane, PCBP2 stabilizes NDUFS1 mRNA to engage NRF2 anti-ferroptosis signaling, caspase-3 cleaves at D255, and the protein's NAD+ output controls ENaCα in alveolar clearance.","evidence":"SIRT3 activation/acetylation IP and BN-PAGE; RIP and pull-down with MI rescue; D255A cleavage-site mutant; knockdown with Olaparib NAD+ rescue","pmids":["40493314","40784311","41422996","40860777"],"confidence":"Medium","gaps":["Interplay among deacetylation, lactylation and glutathionylation untested","ENaCα link is downstream/correlative"]},{"year":2026,"claim":"Refined regulation by import and modification: GL-V9 acts as a molecular glue strengthening MDM2-NDUFS1 cytoplasmic sequestration to trigger the OMA1-DELE1 stress response, K170 lactylation modulates ischemic myocardial injury, and CD147-pSTAT3 transcription of NDUFS1 sustains pancreatic cancer stem-cell metaboloepigenetics.","evidence":"SPR/CETSA/GST pull-down with MDM2 mutagenesis (GL-V9); K170 lactylation mutant in MI/RI model; CSC sorting, ChIP and SIRT1-DNMT1-PAX2 pathway analysis","pmids":["41951044","42134300","41930324"],"confidence":"Medium","gaps":["Lactylation enzyme/eraser not identified","How complex I activity drives nuclear epigenetic signaling mechanistically unresolved"]},{"year":null,"claim":"How the competing import, modification, and degradation inputs (AKAP1, MDM2, SIRT3, glutathionylation, lactylation, RNF43, caspase-3) are integrated to set NDUFS1 abundance and complex I output in a given cell type remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model of competing localization/PTM control","No high-resolution structure of human NDUFS1 within the N-module","Stoichiometry and timing of modifications uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016491","term_label":"oxidoreductase activity","supporting_discovery_ids":[0,1,6]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0,2,11]}],"localization":[{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[5,7,19]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[5,7,20]}],"pathway":[{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[0,1,6,18]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[4,17,20]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[10,12,16]}],"complexes":["Mitochondrial respiratory complex I (N-module)"],"partners":["NDUFV1","MDM2","AKAP1","PHB2","RNF43","PCBP2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P28331","full_name":"NADH-ubiquinone oxidoreductase 75 kDa subunit, mitochondrial","aliases":["Complex I-75kD","CI-75kD"],"length_aa":727,"mass_kda":79.5,"function":"Core subunit of the mitochondrial membrane respiratory chain NADH dehydrogenase (Complex I) which catalyzes electron transfer from NADH through the respiratory chain, using ubiquinone as an electron acceptor (PubMed:30879903, PubMed:31557978). Essential for catalysing the entry and efficient transfer of electrons within complex I (PubMed:31557978). Plays a key role in the assembly and stability of complex I and participates in the association of complex I with ubiquinol-cytochrome reductase complex (Complex III) to form supercomplexes (PubMed:30879903, PubMed:31557978)","subcellular_location":"Mitochondrion inner membrane","url":"https://www.uniprot.org/uniprotkb/P28331/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NDUFS1","classification":"Not Classified","n_dependent_lines":494,"n_total_lines":1208,"dependency_fraction":0.40894039735099336},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"MIF","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NDUFS1","total_profiled":1310},"omim":[{"mim_id":"619701","title":"YOON-BELLEN NEURODEVELOPMENTAL SYNDROME; YOBELN","url":"https://www.omim.org/entry/619701"},{"mim_id":"619382","title":"LEBER-LIKE HEREDITARY OPTIC NEUROPATHY, AUTOSOMAL RECESSIVE 1; LHONAR1","url":"https://www.omim.org/entry/619382"},{"mim_id":"618226","title":"MITOCHONDRIAL COMPLEX I DEFICIENCY, NUCLEAR TYPE 5; MC1DN5","url":"https://www.omim.org/entry/618226"},{"mim_id":"618225","title":"MITOCHONDRIAL COMPLEX I DEFICIENCY, NUCLEAR TYPE 4; MC1DN4","url":"https://www.omim.org/entry/618225"},{"mim_id":"618202","title":"DNAJ/HSP40 HOMOLOG, SUBFAMILY C, MEMBER 30; DNAJC30","url":"https://www.omim.org/entry/618202"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Mitochondria","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"heart muscle","ntpm":170.1},{"tissue":"skeletal muscle","ntpm":210.5},{"tissue":"tongue","ntpm":210.3}],"url":"https://www.proteinatlas.org/search/NDUFS1"},"hgnc":{"alias_symbol":["CI-75k"],"prev_symbol":[]},"alphafold":{"accession":"P28331","domains":[{"cath_id":"3.10.20.740","chopping":"31-122","consensus_level":"high","plddt":96.2978,"start":31,"end":122},{"cath_id":"3.30.70.20","chopping":"137-297","consensus_level":"medium","plddt":96.159,"start":137,"end":297},{"cath_id":"3.40.50.740","chopping":"322-374_521-652","consensus_level":"high","plddt":96.4962,"start":322,"end":652},{"cath_id":"3.40.228.10","chopping":"388-516_659-673","consensus_level":"high","plddt":94.5849,"start":388,"end":673},{"cath_id":"-","chopping":"690-719","consensus_level":"medium","plddt":91.0497,"start":690,"end":719}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P28331","model_url":"https://alphafold.ebi.ac.uk/files/AF-P28331-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P28331-F1-predicted_aligned_error_v6.png","plddt_mean":92.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NDUFS1","jax_strain_url":"https://www.jax.org/strain/search?query=NDUFS1"},"sequence":{"accession":"P28331","fasta_url":"https://rest.uniprot.org/uniprotkb/P28331.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P28331/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P28331"}},"corpus_meta":[{"pmid":"11349233","id":"PMC_11349233","title":"Large-scale deletion and point mutations of the nuclear NDUFV1 and NDUFS1 genes in mitochondrial complex I deficiency.","date":"2001","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/11349233","citation_count":218,"is_preprint":false},{"pmid":"16478720","id":"PMC_16478720","title":"Dysfunctions of cellular oxidative metabolism in patients with mutations in the NDUFS1 and NDUFS4 genes of complex I.","date":"2006","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16478720","citation_count":127,"is_preprint":false},{"pmid":"32072193","id":"PMC_32072193","title":"Akap1 deficiency exacerbates diabetic cardiomyopathy in mice by NDUFS1-mediated mitochondrial dysfunction and apoptosis.","date":"2020","source":"Diabetologia","url":"https://pubmed.ncbi.nlm.nih.gov/32072193","citation_count":95,"is_preprint":false},{"pmid":"35817848","id":"PMC_35817848","title":"Cardiac-specific overexpression of Ndufs1 ameliorates cardiac dysfunction after myocardial infarction by alleviating mitochondrial dysfunction and apoptosis.","date":"2022","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35817848","citation_count":79,"is_preprint":false},{"pmid":"36658121","id":"PMC_36658121","title":"PHB2 promotes colorectal cancer cell proliferation and tumorigenesis through NDUFS1-mediated oxidative phosphorylation.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/36658121","citation_count":67,"is_preprint":false},{"pmid":"30879903","id":"PMC_30879903","title":"MDM2 Integrates Cellular Respiration and Apoptotic Signaling through NDUFS1 and the Mitochondrial Network.","date":"2019","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/30879903","citation_count":66,"is_preprint":false},{"pmid":"15824269","id":"PMC_15824269","title":"Leigh syndrome associated with mitochondrial complex I deficiency due to a novel mutation in the NDUFS1 gene.","date":"2005","source":"Archives of neurology","url":"https://pubmed.ncbi.nlm.nih.gov/15824269","citation_count":64,"is_preprint":false},{"pmid":"20382551","id":"PMC_20382551","title":"Novel mutations in the NDUFS1 gene cause low residual activities in human complex I deficiencies.","date":"2010","source":"Molecular genetics and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/20382551","citation_count":55,"is_preprint":false},{"pmid":"31557978","id":"PMC_31557978","title":"Mutations in NDUFS1 Cause Metabolic Reprogramming and Disruption of the Electron Transfer.","date":"2019","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/31557978","citation_count":53,"is_preprint":false},{"pmid":"21203893","id":"PMC_21203893","title":"Progressive cavitating leukoencephalopathy associated with respiratory chain complex I deficiency and a novel mutation in NDUFS1.","date":"2011","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/21203893","citation_count":41,"is_preprint":false},{"pmid":"23788428","id":"PMC_23788428","title":"TNFα-induced lysosomal membrane permeability is downstream of MOMP and triggered by caspase-mediated NDUFS1 cleavage and ROS formation.","date":"2013","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/23788428","citation_count":40,"is_preprint":false},{"pmid":"33763166","id":"PMC_33763166","title":"Ndufs1 Deficiency Aggravates the Mitochondrial Membrane Potential Dysfunction in Pressure Overload-Induced Myocardial Hypertrophy.","date":"2021","source":"Oxidative medicine and cellular longevity","url":"https://pubmed.ncbi.nlm.nih.gov/33763166","citation_count":36,"is_preprint":false},{"pmid":"28063846","id":"PMC_28063846","title":"Systematic Expression Analysis of Mitochondrial Complex I Identifies NDUFS1 as a Biomarker in Clear-Cell Renal-Cell Carcinoma.","date":"2016","source":"Clinical genitourinary cancer","url":"https://pubmed.ncbi.nlm.nih.gov/28063846","citation_count":32,"is_preprint":false},{"pmid":"25615419","id":"PMC_25615419","title":"Broad phenotypic variability in patients with complex I deficiency due to mutations in NDUFS1 and NDUFV1.","date":"2015","source":"Mitochondrion","url":"https://pubmed.ncbi.nlm.nih.gov/25615419","citation_count":31,"is_preprint":false},{"pmid":"25354934","id":"PMC_25354934","title":"Genetic variant in NDUFS1 gene is associated with schizophrenia and negative symptoms in Han Chinese.","date":"2014","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25354934","citation_count":21,"is_preprint":false},{"pmid":"37644092","id":"PMC_37644092","title":"Loss of NDUFS1 promotes gastric cancer progression by activating the mitochondrial ROS-HIF1α-FBLN5 signaling pathway.","date":"2023","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/37644092","citation_count":19,"is_preprint":false},{"pmid":"33978320","id":"PMC_33978320","title":"MiR-3130-5p is an intermediate modulator of 2q33 and influences the invasiveness of lung adenocarcinoma by targeting NDUFS1.","date":"2021","source":"Cancer medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33978320","citation_count":15,"is_preprint":false},{"pmid":"24952175","id":"PMC_24952175","title":"A homozygous mutation in the NDUFS1 gene presents with a mild cavitating leukoencephalopathy.","date":"2014","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/24952175","citation_count":14,"is_preprint":false},{"pmid":"38697493","id":"PMC_38697493","title":"Targeted degradation of NDUFS1 by agrimol B promotes mitochondrial ROS accumulation and cytotoxic autophagy arrest in hepatocellular carcinoma.","date":"2024","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38697493","citation_count":13,"is_preprint":false},{"pmid":"36290766","id":"PMC_36290766","title":"Conditions Conducive to the Glutathionylation of Complex I Subunit NDUFS1 Augment ROS Production following the Oxidation of Ubiquinone Linked Substrates, Glycerol-3-Phosphate and Proline.","date":"2022","source":"Antioxidants (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/36290766","citation_count":12,"is_preprint":false},{"pmid":"36402252","id":"PMC_36402252","title":"Down regulation of NDUFS1 is involved in the progression of parenteral-nutrition-associated liver disease by increasing Oxidative stress.","date":"2022","source":"The Journal of nutritional biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/36402252","citation_count":11,"is_preprint":false},{"pmid":"38988031","id":"PMC_38988031","title":"m6A methylation of RNF43 inhibits the progression of endometriosis through regulating oxidative phosphorylation via NDUFS1.","date":"2024","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/38988031","citation_count":10,"is_preprint":false},{"pmid":"40493314","id":"PMC_40493314","title":"Berberine dissociates mitochondrial complex I by SIRT3-dependent deacetylation of NDUFS1 to improve hepatocellular glucose and lipid metabolism.","date":"2025","source":"Science China. Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40493314","citation_count":9,"is_preprint":false},{"pmid":"34885151","id":"PMC_34885151","title":"Proteomic Analysis Identifies NDUFS1 and ATP5O as Novel Markers for Survival Outcome in Prostate Cancer.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34885151","citation_count":5,"is_preprint":false},{"pmid":"40617306","id":"PMC_40617306","title":"Naringin attenuates myocardial ischemia-reperfusion injury by promoting mitochondrial translocation of NDUFS1 and suppressing cardiac microvascular endothelial cell ferroptosis.","date":"2025","source":"The Journal of nutritional biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/40617306","citation_count":4,"is_preprint":false},{"pmid":"40880646","id":"PMC_40880646","title":"Mechanisms of sorafenib-induced cardiotoxicity: ER stress induces upregulation of ATF3, leading to downregulation of NDUFS1 expression and mitochondrial dysfunction.","date":"2025","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/40880646","citation_count":4,"is_preprint":false},{"pmid":"36462614","id":"PMC_36462614","title":"Analysis of compound heterozygous and homozygous mutations found in peripheral subunits of human respiratory Complex I, NDUFS1, NDUFS2, NDUFS8 and NDUFV1, by modeling in the E. coli enzyme.","date":"2022","source":"Mitochondrion","url":"https://pubmed.ncbi.nlm.nih.gov/36462614","citation_count":4,"is_preprint":false},{"pmid":"38720274","id":"PMC_38720274","title":"Association between NDUFS1 from urinary extracellular vesicles and decreased differential renal function in children with ureteropelvic junction obstruction.","date":"2024","source":"BMC nephrology","url":"https://pubmed.ncbi.nlm.nih.gov/38720274","citation_count":2,"is_preprint":false},{"pmid":"36042640","id":"PMC_36042640","title":"Lip cyanosis as the first symptom of Leigh syndrome associated with mitochondrial complex I deficiency due to a compound heterozygous NDUFS1 mutation: A case report.","date":"2022","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36042640","citation_count":2,"is_preprint":false},{"pmid":"40784311","id":"PMC_40784311","title":"PCBP2 alleviates myocardial infarction by inhibiting cardiomyocyte ferroptosis via the NDUFS1/NRF2 pathway.","date":"2025","source":"Molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/40784311","citation_count":1,"is_preprint":false},{"pmid":"40860777","id":"PMC_40860777","title":"NDUFS1 upregulates ENaCα by NAD+ to promote alveolar fluid clearance in acute lung injury.","date":"2025","source":"International journal of medical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40860777","citation_count":1,"is_preprint":false},{"pmid":"36403546","id":"PMC_36403546","title":"Generation of an induced pluripotent stem cell line (IUFi002-A) from a Leigh syndrome patient carrying mutations in the NDUFS1 gene.","date":"2022","source":"Stem cell research","url":"https://pubmed.ncbi.nlm.nih.gov/36403546","citation_count":1,"is_preprint":false},{"pmid":"41422996","id":"PMC_41422996","title":"ER-localized ERO1α and caspase-3-mediated cleavage of mitochondrial NDUFS1 drives trichothecene-induced ROS accumulation in liver.","date":"2025","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41422996","citation_count":1,"is_preprint":false},{"pmid":"33751534","id":"PMC_33751534","title":"[Compound heterozygous NDUFS1 variants identified in a Chinese pedigree affected with mitochondrial respiratory chain complex I deficiency].","date":"2021","source":"Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/33751534","citation_count":1,"is_preprint":false},{"pmid":"42040813","id":"PMC_42040813","title":"Expression Study of NDUFS1, NDUFV1, and NDUFV2 in Schizophrenia and Paranoid Personality Disorder : Role of Mitochondrial Complex I in SCZ and PPD.","date":"2022","source":"Galen medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/42040813","citation_count":0,"is_preprint":false},{"pmid":"40794258","id":"PMC_40794258","title":"Deep learning-based radiolabelled compound-protein interaction prediction for NDUFS1-targeting radiopharmaceutical discovery.","date":"2025","source":"EJNMMI research","url":"https://pubmed.ncbi.nlm.nih.gov/40794258","citation_count":0,"is_preprint":false},{"pmid":"41672192","id":"PMC_41672192","title":"Role of NDUFS1 in mitochondrial dysfunction and oxidative stress in glaucomatous retinal ganglion cells.","date":"2026","source":"Experimental eye research","url":"https://pubmed.ncbi.nlm.nih.gov/41672192","citation_count":0,"is_preprint":false},{"pmid":"42134300","id":"PMC_42134300","title":"Piceatannol-3'-O-β-d-glucopyranoside mitigates myocardial ischemia-reperfusion injury by inhibiting ferroptosis through the regulation of NDUFS1 lactylation via metabolic reprogramming.","date":"2026","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/42134300","citation_count":0,"is_preprint":false},{"pmid":"41930324","id":"PMC_41930324","title":"NDUFS1-Mediated Mitochondrial Complex I Activity Maintains Pancreatic Cancer Stemness by Promoting PAX2 Hypomethylation.","date":"2026","source":"MedComm","url":"https://pubmed.ncbi.nlm.nih.gov/41930324","citation_count":0,"is_preprint":false},{"pmid":"42070526","id":"PMC_42070526","title":"Generation of an iPSC line IUFi004-A-13 with homozygous NDUFS1 mutation for the study of Leigh syndrome.","date":"2026","source":"Stem cell research","url":"https://pubmed.ncbi.nlm.nih.gov/42070526","citation_count":0,"is_preprint":false},{"pmid":"41951044","id":"PMC_41951044","title":"GL-V9 disrupts the mitochondrial homeostasis and triggers the integrated stress response by promoting the binding of cytosolic MDM2 with NDUFS1 in colorectal cancer.","date":"2026","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/41951044","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.15.676326","title":"Long non-coding RNA Cerox1 targets components of the mitochondrial electron transport chain to regulate the memory impairment caused by sleep deprivation","date":"2025-09-17","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.15.676326","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.07.08.663799","title":"NDUFS2 (NADH:Ubiquinone oxidoreductase core subunit S2) in Mitochondrial Electron Transport Chain Complex I is Critical to Oxygen Responsiveness of Human Ductus Arteriosus Smooth Muscle Cells","date":"2025-07-14","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.08.663799","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.06.26.659534","title":"EGFR Mutation Subtypes Modulate Distinct Metabolic Profiles and Clinical Outcomes in Lung Cancer: A Retrospective Analysis","date":"2025-06-27","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.26.659534","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.07.15.25331558","title":"Genetically informed drug target prioritisation and repurposing for major depressive disorder","date":"2025-07-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.07.15.25331558","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":25187,"output_tokens":6650,"usd":0.087655,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15746,"output_tokens":4829,"usd":0.099727,"stage2_stop_reason":"end_turn"},"total_usd":0.187382,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"Point mutations and large-scale deletions in the NDUFS1 gene (encoding the 75-kDa Fe-S subunit of mitochondrial complex I) cause isolated complex I deficiency in humans, establishing NDUFS1 as a nuclear-encoded structural subunit required for complex I activity.\",\n      \"method\": \"Denaturing HPLC and direct cDNA sequencing of NDUFS1 in patient fibroblasts; biochemical respiratory chain enzyme assays\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct sequencing plus biochemical activity assays in patient-derived cells, single study but multiple mutations and patients\",\n      \"pmids\": [\"11349233\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"A C1564A missense mutation (Q522K) in NDUFS1 reduces the level of mature complex I, markedly inhibits NADH-ubiquinone oxidoreductase activity, causes accumulation of mitochondrial H2O2 and superoxide, decreases mitochondrial potential, and leads to glutathione depletion; ROS increase was not observed in the NDUFS4 mutant, demonstrating a specific role for the Fe-S NDUFS1 subunit in electron transfer and ROS generation.\",\n      \"method\": \"Biochemical assays in patient fibroblasts: complex I activity measurement, ROS detection (H2O2, O2•−), mitochondrial membrane potential measurement, glutathione quantification, glutathione peroxidase activity assay; dibutyryl-cAMP rescue experiment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal biochemical methods in patient cells with mechanistic rescue experiment, single lab\",\n      \"pmids\": [\"16478720\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Novel NDUFS1 mutations (including a premature stop, amino acid substitutions, and a single-amino-acid deletion) cause decreased complex I amount and activity and a disturbed complex I assembly pattern in patient fibroblasts, establishing NDUFS1 as required for proper assembly and stability of mitochondrial complex I.\",\n      \"method\": \"NDUFS1 gene sequencing in patient fibroblasts; complex I activity assay; Blue Native PAGE assembly analysis\",\n      \"journal\": \"Molecular genetics and metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical activity and native PAGE assembly assays across three independent patients, single lab\",\n      \"pmids\": [\"20382551\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"A homozygous p.Thr595Ala mutation in NDUFS1 causes severe reduction of complex I enzyme activity in muscle and complex I dysfunction in a Neurospora crassa insertional mutagenesis model and in patient fibroblasts grown in galactose, providing cross-species genetic evidence that NDUFS1 is functionally conserved and essential for complex I activity.\",\n      \"method\": \"Muscle biopsy complex I enzyme activity assay; Neurospora crassa insertional mutagenesis model; patient fibroblasts grown in galactose (stress condition to unmask OXPHOS defect)\",\n      \"journal\": \"Neurogenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — orthologous model organism mutagenesis plus patient biochemistry, single study\",\n      \"pmids\": [\"21203893\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Caspase-3 cleaves the p75 NDUFS1 subunit of respiratory complex I downstream of MOMP during TNFα+cycloheximide-induced apoptosis; this cleavage drives ROS formation, which then triggers lysosomal membrane permeability (LMP) and cathepsin release, amplifying apoptosis. A caspase-non-cleavable p75 mutant prevented LMP, confirming the NDUFS1 cleavage event as mechanistically causal.\",\n      \"method\": \"Genetic epistasis with Bax/Bak, Apaf-1, caspase-9, caspase-3/7 double-knockout cells; caspase-non-cleavable NDUFS1 mutant expression; MitoQ antioxidant rescue; LMP and cathepsin release assays; ROS measurement\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — mutagenesis of cleavage site combined with genetic KO epistasis and pharmacological rescue across multiple orthogonal readouts\",\n      \"pmids\": [\"23788428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"MDM2 directly binds NDUFS1 via its amino-terminal region (aa 1–101), sequesters it in the cytoplasm, prevents its mitochondrial localization, and thereby destabilizes complex I and respiratory supercomplexes, leading to decreased mitochondrial respiration, oxidative stress, and commitment to the mitochondrial apoptosis pathway in a p53-independent manner.\",\n      \"method\": \"Complementary biochemical (Co-IP, pull-down), organellar fractionation, and cellular approaches; MDM2 amino-terminal truncation mapping; supercomplex analysis by BN-PAGE; Drosophila and murine transgenic Mdm2 models; oxygen consumption rate assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct binding demonstrated by pull-down with domain mapping, replicated in two animal models, multiple orthogonal functional readouts\",\n      \"pmids\": [\"30879903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Biallelic NDUFS1 mutations decrease the stability of the entire N-module of complex I and disrupt electron transfer between two iron-sulfur clusters within NDUFS1, causing metabolic reprogramming including TCA cycle inhibitory feedback and elevated reactive oxygen species stress.\",\n      \"method\": \"Proteome and metabolome profiling of patient-derived cells; structural inference from iron-sulfur cluster positions; comparison with a second CI gene mutation\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteomics plus metabolomics in patient cells with mechanistic interpretation from structural context, single lab\",\n      \"pmids\": [\"31557978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AKAP1 interacts with NDUFS1 (identified by immunoprecipitation and mass spectrometry) and is required for translocation of NDUFS1 from the cytosol to mitochondria; AKAP1 deficiency prevents this translocation, inhibits complex I activity, reduces ATP production, increases mitochondrial ROS, and exacerbates cardiomyocyte apoptosis. Restoration of AKAP1 rescues mitochondrial NDUFS1 localization and cardiac function.\",\n      \"method\": \"Co-immunoprecipitation and LC-MS/MS; Akap1-KO mice with STZ-induced diabetes; AAV9-Akap1 cardiac overexpression rescue; echocardiography; complex I activity assay; ROS measurement; subcellular fractionation\",\n      \"journal\": \"Diabetologia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP/MS plus in vivo KO and AAV rescue with multiple functional readouts establishing the AKAP1–NDUFS1 mitochondrial import mechanism\",\n      \"pmids\": [\"32072193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NDUFS1 knockdown in cardiomyocytes decreases mitochondrial DNA content, mitochondrial membrane potential, and mitochondrial mass while increasing mitochondrial ROS production; Ndufs1 overexpression reverses Ang II-induced cardiomyocyte hypertrophy phenotypes, establishing a direct role for NDUFS1 in maintaining mitochondrial membrane potential in cardiomyocytes.\",\n      \"method\": \"siRNA knockdown and overexpression of Ndufs1 in rat cardiomyocytes; Ang II hypertrophy model; MMP measurement (JC-1), mtDNA content, mitochondrial mass, and ROS assays\",\n      \"journal\": \"Oxidative medicine and cellular longevity\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss- and gain-of-function with multiple mitochondrial phenotype readouts, single lab\",\n      \"pmids\": [\"33763166\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"PHB2 directly interacts with NDUFS1 (identified by Co-IP and mass spectrometry) and co-localizes with it in mitochondria; this interaction facilitates NDUFS1 binding to NDUFV1, stabilizes complex I, and enhances complex I activity, thereby elevating oxidative phosphorylation levels in colorectal cancer cells.\",\n      \"method\": \"Co-immunoprecipitation and mass spectrometry; confocal co-localization; complex I activity assay after PHB2 knockdown or overexpression; PHB2 KD combined with PHB2 OE rescue\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP/MS plus co-localization and functional complex I activity assay, single lab\",\n      \"pmids\": [\"36658121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Glutathionylation of NDUFS1 within complex I (induced by disulfiram) increases mitochondrial superoxide/H2O2 production during reverse electron transfer from the ubiquinone pool via substrates glycerol-3-phosphate and proline; deglutathionylation of NDUFS1 by reducing agents restores normal complex I activity and decreases ROS production.\",\n      \"method\": \"Immunocapture of complex I from liver mitochondria; disulfiram-induced glutathionylation; site-specific inhibitors for complex I, III, GPD, PRODH; ROS measurement; reducing-agent reversal of glutathionylation\",\n      \"journal\": \"Antioxidants (Basel, Switzerland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro mitochondrial assay with immunocapture, pharmacological dissection of ROS sites, and reversible modification, single lab\",\n      \"pmids\": [\"36290766\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Mutations at NDUFS1-corresponding positions in the homologous E. coli nuoG subunit reduce NADH oxidase activity and disrupt complex I assembly (assessed by co-immunoprecipitation and time-delayed expression assays), and many map to subunit interfaces; compound heterozygote modeling identified which mutation in a pair is more deleterious.\",\n      \"method\": \"Site-directed mutagenesis in E. coli nuoG (NDUFS1 ortholog); membrane vesicle NADH oxidase activity assay; co-immunoprecipitation assembly assay; time-delayed expression assay; alanine substitution series\",\n      \"journal\": \"Mitochondrion\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution in bacterial model with mutagenesis and assembly assays, single lab, ortholog model\",\n      \"pmids\": [\"36462614\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Reduction of NDUFS1 in gastric cancer cells activates the mitochondrial ROS–HIF1α signaling pathway, upregulating FBLN5 (a transcriptional target of HIF1α), thereby promoting cancer cell proliferation, migration, and invasion; NDUFS1 overexpression suppresses this pathway and inhibits tumor growth in vivo.\",\n      \"method\": \"Confocal microscopy for NDUFS1 subcellular localization and mROS measurement; CCK-8, colony formation, transwell assays; mouse xenograft model; western blot and IHC for pathway components\",\n      \"journal\": \"British journal of cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss- and gain-of-function with in vivo xenograft, pathway component measurement, and localization imaging, single lab\",\n      \"pmids\": [\"37644092\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Agrimol B causes caspase-3-mediated degradation of NDUFS1 protein, leading to mitochondrial ROS accumulation, autophagosome-lysosome fusion blockade (autophagy arrest), and HCC cell growth inhibition; NDUFS1 overexpression partially restores mitochondrial ROS levels and reverses autophagy arrest induced by agrimol B.\",\n      \"method\": \"NDUFS1 overexpression rescue experiment; caspase-3 activity assay; mROS measurement; autophagosome accumulation assay; in vitro and PDX in vivo models\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic rescue with NDUFS1 OE and caspase-3 pathway dissection, in vivo PDX validation, single lab\",\n      \"pmids\": [\"38697493\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF43 (an E3 ubiquitin ligase) directly interacts with NDUFS1 and promotes its ubiquitination and proteasomal degradation, reducing oxidative phosphorylation activity; NDUFS1 is thus a downstream target of RNF43 in endometrial stromal cells.\",\n      \"method\": \"Co-immunoprecipitation demonstrating RNF43–NDUFS1 interaction; ubiquitination assay; NDUFS1 knockdown phenocopy of RNF43 overexpression; OXPHOS activity measurement\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP establishing interaction plus ubiquitination assay and functional rescue, single lab\",\n      \"pmids\": [\"38988031\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Berberine directly binds and activates SIRT3, which deacetylates NDUFS1 (the catalytic subunit in the N-module of complex I), causing dissociation of complex I from the mitochondrial membrane; this selectively and reversibly reduces complex I abundance and OXPHOS activity in hepatocytes, improving glucose and lipid metabolism.\",\n      \"method\": \"In vivo oral administration followed by mitochondrial isolation; SIRT3 activation assay; acetylation state of NDUFS1 measured by IP; complex I dissociation by BN-PAGE; oxygen consumption rate; glucose/lipid metabolic readouts\",\n      \"journal\": \"Science China. Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding of berberine to SIRT3 linked to NDUFS1 deacetylation and complex I dissociation with multiple orthogonal readouts, single lab\",\n      \"pmids\": [\"40493314\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PCBP2 binds NDUFS1 mRNA (verified by RNA-immunoprecipitation and RNA-protein pull-down), stabilizes it, and promotes NDUFS1 protein expression; increased NDUFS1 in turn activates NRF2 nuclear translocation, inhibiting cardiomyocyte ferroptosis during myocardial infarction.\",\n      \"method\": \"RNA-immunoprecipitation (RIP); RNA-protein pull-down; NDUFS1 overexpression and PCBP2 overexpression experiments; NRF2 nuclear translocation assay; ferroptosis markers; in vivo MI mouse model with LV-PCBP2/LV-NDUFS1\",\n      \"journal\": \"Molecular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RIP and pull-down establishing RNA-protein interaction, plus in vivo rescue, single lab\",\n      \"pmids\": [\"40784311\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Caspase-3 cleaves NDUFS1 at residue D255; mutation D255A abolishes this cleavage and attenuates ROS accumulation and mitochondrial dysfunction induced by trichothecene mycotoxins, confirming that caspase-3-mediated NDUFS1 cleavage disrupts electron transport and amplifies mitochondrial ROS in a positive feedback loop with ERO1α-mediated ER oxidative stress.\",\n      \"method\": \"In vivo and in vitro mycotoxin exposure models; caspase-3 inhibition and siRNA knockdown; NDUFS1 D255A cleavage-site mutant expression; ROS measurement; mitochondrial function assays\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct site mutagenesis (D255A) preventing caspase-3 cleavage with functional rescue in both in vitro and in vivo models, single lab but two orthogonal approaches\",\n      \"pmids\": [\"41422996\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NDUFS1 deficiency in alveolar epithelial cells reduces complex I activity, impairs NAD+ production, and increases ROS, which in turn decreases ENaCα expression and impairs alveolar fluid clearance; supplementing NAD+ via Olaparib restores ENaCα levels and alleviates acute lung injury phenotypes caused by NDUFS1 deficiency.\",\n      \"method\": \"NDUFS1 knockdown in alveolar epithelial cells; complex I activity assay; NAD+ measurement; ROS assay; ENaCα expression; Olaparib-mediated NAD+ supplementation rescue; ALI mouse models\",\n      \"journal\": \"International journal of medical sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KD plus pharmacological rescue establishing NAD+/ENaCα as mechanistic mediators downstream of NDUFS1, single lab\",\n      \"pmids\": [\"40860777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Naringin facilitates translocation of NDUFS1 from the cytosol to mitochondria in cardiac microvascular endothelial cells during hypoxia-reoxygenation injury; this mitochondrial import of NDUFS1 restores mitochondrial function, reduces ROS, and suppresses ferroptosis via the IRF3/SLC7A11/GPX4 axis.\",\n      \"method\": \"Immunofluorescence and subcellular fractionation to track NDUFS1 localization; proteomic analysis; molecular docking and molecular dynamics; ferroptosis marker assays; in vivo MI/RI rat model\",\n      \"journal\": \"The Journal of nutritional biochemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization by immunofluorescence with functional correlation but no direct interaction mapping; single lab, single study\",\n      \"pmids\": [\"40617306\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"GL-V9 binds the MDM2 amino-terminal domain (aa 1–101) and acts as a molecular glue that facilitates MDM2–NDUFS1 interaction in the cytoplasm, preventing NDUFS1 mitochondrial localization, inhibiting complex I formation, disrupting mitochondrial homeostasis, and activating the OMA1-DELE1 integrated stress response to induce apoptosis, in a p53-independent manner.\",\n      \"method\": \"GST pull-down assay; cellular thermal shift assay (CETSA); surface plasmon resonance (SPR); immunofluorescence for NDUFS1 mitochondrial localization; MDM2 amino acid mutation mapping; mitochondrial membrane potential, superoxide, ATP, OCR assays; OMA1-DELE1 pathway readout\",\n      \"journal\": \"Journal of advanced research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — direct binding confirmed by SPR, CETSA, GST pull-down, and amino acid mutagenesis; NDUFS1 localization and complex I function validated by multiple orthogonal assays; single lab\",\n      \"pmids\": [\"41951044\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"NDUFS1-mediated complex I activity maintains pancreatic cancer stem cell stemness and tumorigenicity; mechanistically, CD147 promotes pSTAT3Tyr705-mediated NDUFS1 transcription, and NDUFS1 initiates SIRT1-DNMT1 metaboloepigenetic signaling that reduces PAX2 promoter methylation, increasing PAX2 expression to sustain stemness.\",\n      \"method\": \"ALDH+ CSC sorting and tumorsphere assay; complex I activity assays; NDUFS1 KD/OE; ChIP for PAX2 promoter methylation; DNMT1/SIRT1 pathway analysis; CD147 overexpression; in vivo tumorigenicity assays\",\n      \"journal\": \"MedComm\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal functional assays linking NDUFS1 to epigenetic regulation of stemness, single lab\",\n      \"pmids\": [\"41930324\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"NDUFS1 K170 lactylation (induced during ischemia-reperfusion) impairs the cardioprotective effects of PG; overexpression of NDUFS1 K170 lactylation diminished PG-mediated improvement of MIRI, establishing a specific lysine lactylation site on NDUFS1 as a post-translational modification that modulates mitochondrial function and ferroptosis in myocardial injury.\",\n      \"method\": \"Multi-omics (metabolomics, proteomics); overexpression of NDUFS1 K170 lactylation mutant; PDK4 overexpression; in vivo rat MI/RI model; GPX4, ACSL4, PDK4 pathway readouts\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-specific lactylation mutant with in vivo functional readout, single lab, single study\",\n      \"pmids\": [\"42134300\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"ATF3, upregulated downstream of the ER stress PERK-eIF2α-ATF4 pathway by sorafenib, negatively regulates NDUFS1 expression; siRNA silencing of ATF3 partially restores mitochondrial function impaired by sorafenib, defining an ATF3→NDUFS1 regulatory axis in sorafenib-induced cardiotoxicity.\",\n      \"method\": \"Transcriptomic and proteomic profiling; ATF3 siRNA knockdown; Western blot validation; mitochondrial function assays; ER stress inhibitor (GSK2606414) rescue; H9C2 cell model and in vivo rat model\",\n      \"journal\": \"Frontiers in pharmacology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — ATF3 KD with functional rescue but mechanistic link to NDUFS1 relies primarily on proteomic screening without direct ATF3–NDUFS1 promoter or binding assay, single lab\",\n      \"pmids\": [\"40880646\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NDUFS1 (CI-75k) encodes the 75-kDa iron-sulfur subunit that forms the core of the N-module of mitochondrial respiratory chain complex I, where it coordinates iron-sulfur clusters essential for electron transfer from NADH to ubiquinone; it must be imported from the cytosol into mitochondria (a step facilitated by AKAP1 and antagonized by MDM2 sequestration), assembles into complex I through subunit interfaces (where pathogenic mutations disrupt assembly and activity), and its activity is regulated post-translationally by glutathionylation (increasing ROS via reverse electron transfer), SIRT3-dependent deacetylation (leading to complex I dissociation), and lactylation at K170; caspase-3 cleaves NDUFS1 during apoptosis to amplify mitochondrial ROS and lysosomal membrane permeability, while binding partners including PHB2, CD147-STAT3, and RNF43 modulate its stability and activity to control oxidative phosphorylation, ROS production, and downstream signaling in diverse cellular contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NDUFS1 encodes the 75-kDa iron-sulfur subunit that forms the catalytic core of the N-module of mitochondrial respiratory complex I, where it coordinates iron-sulfur clusters that relay electrons from NADH toward ubiquinone [#0, #6]. Pathogenic point mutations, deletions, and single-residue substitutions reduce complex I amount and NADH-ubiquinone oxidoreductase activity and disrupt assembly and stability of the entire N-module, causing isolated complex I deficiency in humans; many disease residues map to subunit interfaces and to the electron-transfer path between two internal Fe-S clusters, and loss of activity drives accumulation of mitochondrial superoxide and H2O2, membrane-potential collapse, and glutathione depletion [#0, #1, #2, #6, #11]. Because NDUFS1 is nuclear-encoded, its function depends on import into mitochondria: AKAP1 binds NDUFS1 and is required for its cytosol-to-mitochondria translocation, whereas MDM2 directly binds the protein and sequesters it in the cytoplasm, destabilizing complex I and respiratory supercomplexes and committing cells to mitochondrial apoptosis in a p53-independent manner [#5, #7]. NDUFS1 abundance and activity are further tuned post-translationally and through protein and RNA partners: glutathionylation increases ROS via reverse electron transfer [#10], SIRT3-dependent deacetylation dissociates complex I from the membrane [#15], K170 lactylation modulates mitochondrial function in ischemic myocardium [#22], RNF43 ubiquitinates it for proteasomal degradation [#14], PHB2 stabilizes the NDUFS1-NDUFV1 interface to enhance OXPHOS [#9], and PCBP2 binds and stabilizes NDUFS1 mRNA [#16]. During apoptosis, caspase-3 cleaves NDUFS1 (at D255) to amplify mitochondrial ROS, which in turn triggers lysosomal membrane permeabilization and ER oxidative stress feedback [#4, #17]. Through these mechanisms NDUFS1 acts as a node controlling oxidative phosphorylation, ROS output, and downstream signaling — including HIF1\\u03b1, NRF2, and ferroptosis-related pathways — in cardiac, epithelial, and cancer contexts [#8, #12, #16, #18, #21].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established NDUFS1 as a nuclear-encoded structural subunit whose mutation directly causes a respiratory chain disease, answering whether the gene is required for complex I function in humans.\",\n      \"evidence\": \"dHPLC and cDNA sequencing of patient fibroblasts plus respiratory chain enzyme assays\",\n      \"pmids\": [\"11349233\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not resolve how individual mutations affect the electron-transfer path\", \"No assembly-stage analysis\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Showed the Fe-S NDUFS1 subunit specifically governs both electron transfer and ROS generation, distinguishing it from other complex I subunits whose loss does not raise ROS.\",\n      \"evidence\": \"Biochemical activity, ROS, membrane-potential and glutathione assays in Q522K patient fibroblasts with cAMP rescue\",\n      \"pmids\": [\"16478720\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Single mutation in one patient line\", \"Mechanism of ROS amplification not at residue resolution\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Demonstrated that NDUFS1 mutations disturb complex I assembly, not just steady-state activity, defining the subunit as required for complex assembly and stability.\",\n      \"evidence\": \"Gene sequencing, activity assays and Blue Native PAGE assembly analysis in three patients\",\n      \"pmids\": [\"20382551\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Assembly intermediates not structurally mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Provided cross-species confirmation that NDUFS1 is functionally conserved and essential, addressing whether human findings reflect a core conserved role.\",\n      \"evidence\": \"Muscle enzymology, Neurospora crassa insertional mutant, and galactose-stressed patient fibroblasts\",\n      \"pmids\": [\"21203893\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single homozygous mutation\", \"No direct biochemical Fe-S cluster measurement\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined a non-respiratory role: caspase-3 cleavage of NDUFS1 during apoptosis amplifies ROS and triggers lysosomal membrane permeabilization, placing the subunit in an apoptotic feed-forward circuit.\",\n      \"evidence\": \"Bax/Bak, Apaf-1, caspase-9, caspase-3/7 KO epistasis, caspase-non-cleavable mutant, and MitoQ rescue\",\n      \"pmids\": [\"23788428\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Exact cleavage residue not defined here\", \"Generality across apoptotic stimuli untested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified MDM2 as a direct cytoplasmic sequestering partner that blocks NDUFS1 mitochondrial import to suppress respiration and promote apoptosis independently of p53.\",\n      \"evidence\": \"Co-IP/pull-down with N-terminal (aa 1-101) domain mapping, BN-PAGE supercomplex analysis, and Drosophila/mouse Mdm2 models\",\n      \"pmids\": [\"30879903\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How MDM2 selects NDUFS1 over other import cargo unclear\", \"No structure of the MDM2-NDUFS1 interface\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linked NDUFS1 mutations to destabilization of the whole N-module and disrupted inter-cluster electron transfer, connecting genotype to metabolic reprogramming.\",\n      \"evidence\": \"Proteome/metabolome profiling of patient cells with structural inference from Fe-S cluster positions\",\n      \"pmids\": [\"31557978\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Electron-transfer disruption inferred, not directly measured\", \"Single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established AKAP1 as a factor required for NDUFS1 import into mitochondria, defining a positive regulator of its localization opposite to MDM2.\",\n      \"evidence\": \"Reciprocal Co-IP/LC-MS/MS, Akap1-KO diabetic mice, AAV9-Akap1 cardiac rescue and fractionation\",\n      \"pmids\": [\"32072193\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether AKAP1 acts at the import machinery directly is unresolved\", \"Interaction interface not mapped\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showed NDUFS1 level controls cardiomyocyte mitochondrial mass, mtDNA content and membrane potential, linking the subunit to hypertrophy phenotypes.\",\n      \"evidence\": \"siRNA knockdown and overexpression in rat cardiomyocytes with Ang II hypertrophy model\",\n      \"pmids\": [\"33763166\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting NDUFS1 to mtDNA content unclear\", \"Single lab, rodent cells\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined several regulators of NDUFS1: PHB2 stabilizes the NDUFS1-NDUFV1 interface to boost OXPHOS, glutathionylation drives reverse-electron-transfer ROS, and ortholog mutagenesis mapped activity/assembly to subunit interfaces.\",\n      \"evidence\": \"Co-IP/MS and complex I assays (PHB2), immunocaptured complex I with reversible glutathionylation, and E. coli nuoG site-directed mutagenesis\",\n      \"pmids\": [\"36658121\", \"36290766\", \"36462614\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Glutathionylation site on NDUFS1 not pinpointed\", \"PHB2 effect shown in one cancer context\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected NDUFS1 loss to mitochondrial ROS-HIF1\\u03b1-FBLN5 signaling that promotes gastric cancer progression, framing NDUFS1 as a tumor suppressor in this context.\",\n      \"evidence\": \"Loss/gain-of-function with xenografts, localization imaging and pathway component blots/IHC\",\n      \"pmids\": [\"37644092\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct HIF1\\u03b1 induction kinetics not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showed NDUFS1 degradation routes converge on ROS and downstream cell-fate outcomes: caspase-3-driven degradation arrests autophagy in liver cancer, and RNF43-mediated ubiquitination lowers OXPHOS in endometrial stroma.\",\n      \"evidence\": \"NDUFS1 overexpression rescue with caspase-3 dissection (agrimol B), and Co-IP plus ubiquitination assay (RNF43)\",\n      \"pmids\": [\"38697493\", \"38988031\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"RNF43 ubiquitination site on NDUFS1 not mapped\", \"Pathway specificity across tissues unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Expanded the post-translational and transcriptional control of NDUFS1: SIRT3 deacetylation dissociates complex I from the membrane, PCBP2 stabilizes NDUFS1 mRNA to engage NRF2 anti-ferroptosis signaling, caspase-3 cleaves at D255, and the protein's NAD+ output controls ENaC\\u03b1 in alveolar clearance.\",\n      \"evidence\": \"SIRT3 activation/acetylation IP and BN-PAGE; RIP and pull-down with MI rescue; D255A cleavage-site mutant; knockdown with Olaparib NAD+ rescue\",\n      \"pmids\": [\"40493314\", \"40784311\", \"41422996\", \"40860777\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interplay among deacetylation, lactylation and glutathionylation untested\", \"ENaC\\u03b1 link is downstream/correlative\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Refined regulation by import and modification: GL-V9 acts as a molecular glue strengthening MDM2-NDUFS1 cytoplasmic sequestration to trigger the OMA1-DELE1 stress response, K170 lactylation modulates ischemic myocardial injury, and CD147-pSTAT3 transcription of NDUFS1 sustains pancreatic cancer stem-cell metaboloepigenetics.\",\n      \"evidence\": \"SPR/CETSA/GST pull-down with MDM2 mutagenesis (GL-V9); K170 lactylation mutant in MI/RI model; CSC sorting, ChIP and SIRT1-DNMT1-PAX2 pathway analysis\",\n      \"pmids\": [\"41951044\", \"42134300\", \"41930324\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Lactylation enzyme/eraser not identified\", \"How complex I activity drives nuclear epigenetic signaling mechanistically unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the competing import, modification, and degradation inputs (AKAP1, MDM2, SIRT3, glutathionylation, lactylation, RNF43, caspase-3) are integrated to set NDUFS1 abundance and complex I output in a given cell type remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model of competing localization/PTM control\", \"No high-resolution structure of human NDUFS1 within the N-module\", \"Stoichiometry and timing of modifications uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016491\", \"supporting_discovery_ids\": [0, 1, 6]},\n      {\"term_id\": \"GO:0009055\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0, 2, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [5, 7, 19]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [5, 7, 20]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [0, 1, 6, 18]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [4, 17, 20]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [10, 12, 16]}\n    ],\n    \"complexes\": [\"Mitochondrial respiratory complex I (N-module)\"],\n    \"partners\": [\"NDUFV1\", \"MDM2\", \"AKAP1\", \"PHB2\", \"RNF43\", \"PCBP2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"tie","faith_supported":6,"faith_total":6,"faith_pct":100.0}}