{"gene":"MED25","run_date":"2026-04-28T18:30:28","timeline":{"discoveries":[{"year":2004,"finding":"Human MED25 (ARC92) was biochemically purified as a 92-kDa subunit of the ARC/Mediator coactivator complex that is specifically targeted by the activation domain of the VP16 transactivator; VP16 activation domain associates with ARC92 in vitro and in vivo, and siRNA-mediated knockdown of ARC92 selectively inhibits Gal4-VP16 gene activation.","method":"Affinity chromatography with VP16 activation domain, peptide microsequencing, in vitro and in vivo binding assays, siRNA knockdown with reporter gene assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1–2 — biochemical purification, in vitro and in vivo binding, functional siRNA validation; single rigorous study with multiple orthogonal methods","pmids":["14983011"],"is_preprint":false},{"year":2007,"finding":"Human MED25 associates with retinoic acid (RA)-bound RAR through its C-terminal LXXLL (NR box) motif, increases RAR/RXR-mediated transcription, and shows intrinsic transcriptional activity through its PTOV domain by direct association with CBP; MED25 is RA-dependently recruited to the RARβ2 promoter ahead of TRAP220, and MED25 knockdown selectively reduces RAR (but not TR) activity.","method":"Co-immunoprecipitation, ChIP, reporter assays with dominant-negative constructs, siRNA knockdown, domain mapping","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (Co-IP, ChIP, siRNA, reporter assays) in a single rigorous study","pmids":["17641689"],"is_preprint":false},{"year":2010,"finding":"Human Med25 is required for HNF4α association with the Mediator complex and for RNA Pol II recruitment to a subset of HNF4α target gene promoters that selectively regulate drug and lipid metabolism; changes in endogenous Med25 levels alter the composition of the transcriptional complex and Pol II occupancy.","method":"Co-IP, ChIP, siRNA knockdown and overexpression with mRNA expression analysis","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, ChIP, gain- and loss-of-function with defined transcriptional phenotype","pmids":["21135126"],"is_preprint":false},{"year":2011,"finding":"NMR structure of the MED25 activator interaction domain (ACID) reveals a seven-stranded β-barrel framed by three helices; VP16 subdomain H1 and H2 bind to opposite faces of ACID and cooperate during promoter-dependent activated transcription in vitro; the activator-binding ACID faces are functionally required and conserved among higher eukaryotes.","method":"NMR structure determination, mutagenesis, in vitro transcription assay","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 — NMR structure with mutagenesis and in vitro functional validation","pmids":["21378965"],"is_preprint":false},{"year":2010,"finding":"NMR structure of the human MED25 ACID domain reveals a closed β-barrel with seven strands and three α-helices, structurally similar to the SPOC domain-like superfamily; NMR chemical shift mapping shows VP16 H2 interacts with MED25 ACID through one face of the β-barrel defined by strands B4-B7-B6.","method":"NMR structure determination, NMR chemical shift mapping","journal":"Journal of structural biology","confidence":"High","confidence_rationale":"Tier 1 — NMR structure with interaction mapping","pmids":["20974256"],"is_preprint":false},{"year":2011,"finding":"Solution NMR structure of human MED25(391-543) ACID domain shows a β-barrel architecture similar to Ku and SPOC domains; this domain interacts with the acidic transactivation domains of HSV-1 VP16 and VZV IE62.","method":"Solution NMR structure determination, backbone dynamics (15N-{1H} NOEs)","journal":"Journal of structural and functional genomics","confidence":"High","confidence_rationale":"Tier 1 — NMR structure determination with functional context","pmids":["21785987"],"is_preprint":false},{"year":2013,"finding":"MED25 plays a critical role in ATF6α-dependent recruitment of Mediator to ER stress response gene promoters; a specific MED25 domain serves as a docking site on Mediator for the ATF6α transcription activation domain.","method":"Co-IP, domain mapping, ChIP, in vitro binding","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP and domain mapping with ChIP, single lab study","pmids":["23864652"],"is_preprint":false},{"year":2013,"finding":"The N-terminal acidic transactivation domain of PEA3 group ETS transcription factors (PEA3, ERM, ER81) directly contacts the ACID/PTOV domain of MED25; MED25 depletion disrupts ERM-Mediator association in vitro, reduces PEA3-driven MMP-1 expression, and Mediator recruitment; mutations preventing MED25-ERM binding strongly reduce transactivation.","method":"Co-IP, siRNA knockdown, reporter assays, domain mutagenesis, in vitro binding","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (Co-IP, siRNA, mutagenesis, reporter assay, ChIP) with defined functional phenotype","pmids":["23531547"],"is_preprint":false},{"year":2014,"finding":"MED25 is recruited to the CYP2C9 promoter through HNF4α; MED25 influences H3K27 acetylation status—overexpression enriches H3K27ac and CREBBP, while silencing leads to H3K27me3 and increased PRC2 occupancy at the promoter and on HNF4α itself; MED25 induces a permissive chromatin state (FAIRE) required for CYP2C9 transcription.","method":"ChIP, overexpression and siRNA silencing, FAIRE, in vitro transcription reconstitution","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP, FAIRE, in vitro reconstitution, gain and loss of function with epigenetic and transcriptional readouts","pmids":["25391650"],"is_preprint":false},{"year":2014,"finding":"MED25 interacts with HNF4α in pancreatic β-cells through the MED25 LXXLL motif in a ligand-independent manner; this interaction is required for full activation of HNF4α-mediated transcription and glucose-stimulated insulin secretion; MODY mutations at the LXXLL-binding pocket disrupt the interaction and impair insulin secretion via a loss-of-function mechanism.","method":"Yeast two-hybrid, Co-IP, in vitro binding, reporter assays, siRNA/shRNA knockdown, domain mutagenesis","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods with defined cellular phenotype (insulin secretion)","pmids":["22952853"],"is_preprint":false},{"year":2015,"finding":"The Y39C mutation in the MED25 von Willebrand factor type A (VWA) domain, which is responsible for MED25 recruitment into the Mediator complex, dramatically impairs MED25 interaction with the Mediator complex in mammalian cells, as shown by co-immunoprecipitation.","method":"Co-immunoprecipitation of mutant vs. wild-type MED25 with Mediator complex in mammalian cells","journal":"Human genetics","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP with disease-relevant mutation; single lab, single method","pmids":["25792360"],"is_preprint":false},{"year":2015,"finding":"The ERM/ETV5 N-terminal acidic transactivation domain (TAD) folds upon binding to MED25 ACID, forming a fuzzy complex; two aromatic residues (F47 and W57) in ERM TAD mediate binding; mutation Q451 in the VP16 H1 binding pocket of MED25 affects ERM binding; ERM and VP16 H1 share a common binding interface on MED25.","method":"NMR spectroscopy, mutational analysis, competition experiments","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1 — NMR with mutagenesis and competition experiments providing atomic-level detail","pmids":["26130716"],"is_preprint":false},{"year":2017,"finding":"ETV4 activation domain (AD) and DNA-binding domain (DBD) each independently interact with MED25, with the AD binding a single MED25 site and the DBD interacting with three MED25 sites, allowing simultaneous binding; MED25 stimulates ETV4 DNA binding in vitro by relieving autoinhibition; AP1 transcription factors JUN and FOS also contact MED25 at overlapping/distinct sites; genome-wide studies show ETV4 and MED25 co-occupy enhancers in prostate cancer cells.","method":"Binding assays (affinity measurements), in vitro DNA binding assays, ChIP-seq, functional enhancer assays","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1–2 — quantitative binding assays, in vitro functional assay, ChIP-seq with multiple orthogonal methods","pmids":["28728983"],"is_preprint":false},{"year":2016,"finding":"Med25 acts as a coactivator of ligand-activated ERα, interacting with ERα through its C-terminal LXXLL motif after BPA exposure, and is functionally involved in BPA-induced transcriptional regulation of CYP2C9 expression and enzyme activity.","method":"Reporter assays, Co-IP, ChIP, siRNA knockdown, enzyme activity assay","journal":"Environmental toxicology","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP, reporter assay, ChIP; single lab","pmids":["27273787"],"is_preprint":false},{"year":2014,"finding":"Med25 enhances ligand-dependent ERα-mediated transcriptional activation of CYP2C9 and interacts with activated ERα through its C-terminal LXXLL motif.","method":"Reporter assays, Co-IP, domain mapping","journal":"Biochemical pharmacology","confidence":"Medium","confidence_rationale":"Tier 2–3 — Co-IP and reporter assay; single lab, limited methods","pmids":["24960263"],"is_preprint":false},{"year":2018,"finding":"NMR spectroscopy and ITC characterization of the p53 transactivation domain (p53TAD) interaction with MED25 ACID shows that p53TAD2 sequence motif forms an amphipathic α-helix that binds an elongated hydrophobic groove of MED25 ACID; the same conserved mechanism is shared with ERM and VP16 TADs.","method":"NMR chemical shift perturbation, isothermal titration calorimetry, mutagenesis, structural modeling","journal":"Molecules (Basel, Switzerland)","confidence":"High","confidence_rationale":"Tier 1 — NMR, ITC, and mutagenesis with structural model","pmids":["30360415"],"is_preprint":false},{"year":2022,"finding":"RSV NS1 protein interacts with MED25 ACID domain through its C-terminal α3 helix (binding the H2 face of ACID, similar to other transcriptional activation domains such as VP16 and ATF6α), and the globular domain of NS1 also contributes; NS1 can compete with ATF6α TAD for MED25 binding; MED25 knockout enhances RSV replication, suggesting an antiviral role.","method":"Yeast two-hybrid, Co-IP (overexpression and endogenous during infection), NMR, in vitro binding, MED25 KO cell line with viral replication assay","journal":"Journal of virology; Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1–2 — NMR, Co-IP, KO functional assay, competition experiments; independently replicated across two papers","pmids":["36102648","35907573"],"is_preprint":false},{"year":2025,"finding":"X-ray crystal structure of the RSV NS1/MED25 ACID complex reveals NS1 straddles and binds two faces of MED25 ACID simultaneously—distinct from known viral activators; NS1 competes with transcription factor transactivation domains for MED25 binding; ChIP-seq and RNA-seq identify ATF3 as a transcription factor whose Mediator recruitment is affected by NS1/MED25 interaction.","method":"X-ray crystallography, NMR, ChIP-seq, RNA-seq, mutagenesis, in vitro binding","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with mutagenesis, NMR, and genome-wide functional validation","pmids":["40128225"],"is_preprint":false},{"year":2025,"finding":"NS1 α/β core domain binds to MED25 ACID cooperatively with NS1 α3 helix to achieve nanomolar affinity; single amino acid substitutions in NS1 α/β domain (notably E110A) significantly reduce NS1-MED25 affinity and attenuate RSV replication; in MED25-knockdown cells, rRSV replication is further attenuated, and the difference between WT and NS1 mutants is partially lost, indicating NS1-MED25 complex contributes to controlling antiviral responses.","method":"NMR, in vitro binding assays, mutagenesis, recombinant RSV replication assay, MED25 knockdown cells, ISG expression analysis","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1–2 — NMR, mutagenesis, KD cells with viral replication readout; multiple orthogonal methods","pmids":["40920851"],"is_preprint":false},{"year":2021,"finding":"A short binding domain of MED25 (CMIDM) is responsible for interaction with MYC3 (plant ortholog context, but mechanistically relevant to MED25 function); bipartite interaction is critical for stable MYC3-MED25 complex formation; quantitative binding analysis reveals binding affinity order JAZJas < MED25CMIDM < JAZCMID, suggesting a mechanism for activation and negative feedback in jasmonate signaling.","method":"Biochemical binding assays (cell-free system), quantitative affinity measurements, domain mapping","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — quantitative in vitro binding with domain mapping; plant ortholog context","pmids":["34929168"],"is_preprint":false},{"year":2010,"finding":"PTOV1 and MED25 reciprocally regulate RAR transcriptional activity through competitive binding to CBP; PTOV1 antagonizes MED25 by competing for CBP binding and opposing CBP recruitment to RA-responsive gene promoters, affecting RA sensitivity in cancer cells.","method":"Reporter assays, Co-IP, ChIP, overexpression/competition assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2–3 — Co-IP, ChIP, reporter assays; single lab","pmids":["21110951"],"is_preprint":false},{"year":2024,"finding":"Med25 silencing in 3T3-L1 preadipocytes enhances lipid accumulation and super-induces adipogenic master regulators C/EBPα and PPARγ, indicating that Med25 limits adipogenic potential by suppressing levels of these master regulators.","method":"siRNA-mediated silencing, adipogenesis induction, lipid staining, Western blot for C/EBPα and PPARγ","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2–3 — loss-of-function with defined cellular phenotype and molecular pathway; single lab","pmids":["37047128"],"is_preprint":false},{"year":2024,"finding":"A lipopeptidomimetic (LPPM-8) incorporating a medium-chain branched fatty acid binds the H2 face of the MED25 ACID activator interaction domain with ~4 μM Ki (>20-fold improved over unmodified peptide), stabilizes full-length MED25 in cells, and inhibits expression of genes regulated by Med25-activator protein-protein interactions in a triple-negative breast cancer cell model.","method":"In vitro binding assay (Ki determination), cellular proteome stabilization, gene expression analysis in cancer cells","journal":"Angewandte Chemie (International ed. in English)","confidence":"Medium","confidence_rationale":"Tier 2 — quantitative binding, cellular stabilization, and functional gene expression readout; single lab","pmids":["38527936"],"is_preprint":false}],"current_model":"Human MED25 is a structurally characterized subunit of the Mediator coactivator complex whose central ACID domain (a seven-stranded β-barrel) serves as a direct docking platform for the intrinsically disordered acidic transactivation domains of diverse transcription factors—including viral VP16, cellular p53, ETS factors (ETV4/ERM), ATF6α, nuclear receptors (RAR/RXR, HNF4α, ERα) via its C-terminal LXXLL motif, and viral NS1—thereby relaying activator-specific signals to RNA polymerase II; MED25 is required for Pol II recruitment to a selective subset of target gene promoters, influences epigenetic states (H3K27ac vs. H3K27me3) through modulation of HAT (CREBBP) and PRC2 occupancy, and is exploited by RSV NS1 protein, which binds two faces of the ACID domain to compete with cellular transcription factors and suppress antiviral gene expression."},"narrative":{"teleology":[{"year":2004,"claim":"Establishing MED25 as a Mediator subunit and VP16 target resolved how the VP16 activation domain contacts the coactivator complex, identifying MED25 as the specific interface subunit required for VP16-driven transcription.","evidence":"Affinity purification with VP16 activation domain, peptide sequencing, in vivo binding, siRNA knockdown with reporter assay in human cells","pmids":["14983011"],"confidence":"High","gaps":["Whether other transcription factors also dock on MED25 was unknown","Structural basis of the VP16–MED25 interaction was unresolved"]},{"year":2007,"claim":"Demonstrating that MED25 bridges ligand-bound RAR/RXR to Mediator via its LXXLL motif and recruits CBP via its PTOV domain established MED25 as a nuclear receptor coactivator with dual interaction surfaces.","evidence":"Co-IP, ChIP at RARβ2 promoter, reporter assays, siRNA knockdown, domain mapping in human cells","pmids":["17641689"],"confidence":"High","gaps":["How MED25 coordinates with other Mediator-NR contacts (e.g. MED1/TRAP220) was unclear","The structural basis of the LXXLL–NR interaction was not resolved"]},{"year":2010,"claim":"Three independent NMR structures of the MED25 ACID domain revealed a seven-stranded β-barrel architecture with distinct activator-binding faces, providing the first atomic framework for understanding how multiple transcription factors converge on MED25.","evidence":"NMR structure determination, chemical shift mapping of VP16 H2 binding surface","pmids":["20974256","21378965","21785987"],"confidence":"High","gaps":["How VP16 H1 and H2 cooperate on separate ACID faces during transcription was only partially resolved","Whether cellular activators use the same or different binding surfaces was unknown"]},{"year":2010,"claim":"Showing that MED25 is required for HNF4α–Mediator association and Pol II recruitment to a selective subset of drug/lipid metabolism genes established that MED25 functions as a gene-selective coactivator rather than a general Mediator component.","evidence":"Co-IP, ChIP, siRNA knockdown and overexpression with mRNA analysis in human hepatic cells","pmids":["21135126"],"confidence":"High","gaps":["Genome-wide scope of MED25-dependent genes was not defined","How MED25 selectivity for specific HNF4α targets arises was unclear"]},{"year":2013,"claim":"Identifying PEA3-group ETS factors and ATF6α as additional ACID-domain-dependent MED25 clients expanded the activator repertoire beyond VP16 and nuclear receptors, showing MED25 as a shared docking module for diverse acidic transactivation domains.","evidence":"Co-IP, siRNA, reporter assays, mutagenesis, ChIP, domain mapping for ERM/PEA3 and ATF6α","pmids":["23531547","23864652"],"confidence":"High","gaps":["The structural basis for selectivity among different cellular activators at the ACID domain was not resolved","Whether activator competition for MED25 occurs in vivo was untested"]},{"year":2014,"claim":"Demonstrating that MED25 occupancy at target promoters toggles the H3K27ac/H3K27me3 balance by promoting CREBBP recruitment and opposing PRC2 revealed an epigenetic dimension to MED25 coactivator function beyond simple Pol II recruitment.","evidence":"ChIP for histone marks and CREBBP/PRC2, FAIRE, siRNA and overexpression, in vitro transcription reconstitution at CYP2C9 promoter","pmids":["25391650"],"confidence":"High","gaps":["Whether the epigenetic switch is a direct MED25–CREBBP interaction effect or indirect was not fully dissected","Genome-wide extent of MED25-dependent chromatin remodeling was not assessed"]},{"year":2014,"claim":"Linking MED25-HNF4α interaction via the LXXLL motif to glucose-stimulated insulin secretion in β-cells, and showing that MODY-associated HNF4α mutations disrupt this interface, connected MED25 function to a Mendelian disease mechanism.","evidence":"Yeast two-hybrid, Co-IP, reporter assays, siRNA/shRNA knockdown, domain mutagenesis in pancreatic β-cells","pmids":["22952853"],"confidence":"High","gaps":["Whether MED25 variants themselves contribute to diabetes susceptibility was not tested","In vivo validation in animal models was lacking"]},{"year":2015,"claim":"NMR characterization of the ERM TAD–MED25 ACID fuzzy complex, showing that ERM and VP16 H1 share a binding interface, unified the mechanism by which diverse disordered activation domains fold upon docking to a common ACID surface.","evidence":"NMR spectroscopy, mutational analysis, competition experiments","pmids":["26130716"],"confidence":"High","gaps":["Full thermodynamic parameters for all activator–ACID interactions were incomplete","Whether simultaneous multi-activator occupancy is possible was not addressed"]},{"year":2017,"claim":"Discovering that MED25 stimulates ETV4 DNA binding by relieving autoinhibition, and that ETV4 and MED25 co-occupy enhancers genome-wide, revealed an unexpected allosteric function of MED25 beyond passive scaffold bridging.","evidence":"Quantitative binding assays, in vitro DNA binding, ChIP-seq in prostate cancer cells","pmids":["28728983"],"confidence":"High","gaps":["Whether MED25 relieves autoinhibition for other ETS or non-ETS factors was unknown","Structural basis of the allosteric effect was not determined"]},{"year":2018,"claim":"Structural characterization of p53 TAD2 binding to MED25 ACID confirmed that a conserved amphipathic helix mechanism is shared by VP16, ERM, and p53, establishing a unifying structural code for MED25-activator recognition.","evidence":"NMR chemical shift perturbation, ITC, mutagenesis","pmids":["30360415"],"confidence":"High","gaps":["Functional consequences of p53–MED25 interaction for p53 target gene transcription were not assessed in cells","Relative in vivo affinities of competing activators remained unmeasured"]},{"year":2022,"claim":"Showing that RSV NS1 binds MED25 ACID via its α3 helix at the H2 face and competes with ATF6α, with MED25 knockout enhancing viral replication, established MED25 as a host antiviral factor exploited by a pathogen.","evidence":"Yeast two-hybrid, Co-IP (endogenous during infection), NMR, MED25 KO cells with viral replication assay","pmids":["36102648","35907573"],"confidence":"High","gaps":["The full structural basis of the NS1–MED25 complex was not resolved","Which specific antiviral genes are controlled through MED25 was unclear"]},{"year":2025,"claim":"Crystal structure of the NS1–MED25 ACID complex revealed NS1 straddles both ACID faces simultaneously—a binding mode distinct from any known cellular or viral activator—and ChIP-seq/RNA-seq identified ATF3 as a transcription factor whose Mediator recruitment is disrupted by NS1, defining the antiviral gene program controlled through MED25.","evidence":"X-ray crystallography, NMR, ChIP-seq, RNA-seq, mutagenesis, recombinant RSV replication in MED25-knockdown cells","pmids":["40128225","40920851"],"confidence":"High","gaps":["Whether additional transcription factors beyond ATF3 and ATF6α are major in vivo targets of NS1-mediated displacement is not fully mapped","Therapeutic targeting of the NS1–MED25 interface has not been validated in vivo"]},{"year":null,"claim":"A complete in vivo hierarchy of activator competition at the MED25 ACID domain, genome-wide definition of MED25-dependent versus MED25-independent Mediator function, and the structural basis of MED25 integration into the full Mediator complex remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No cryo-EM or crystal structure of MED25 within intact Mediator–Pol II PIC","Genome-wide quantitative model of activator competition for MED25 in physiological contexts is lacking","In vivo animal models with MED25 loss-of-function phenotyping are limited"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,1,2,7,8,9,12]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,6,7,9]}],"localization":[{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[1,8,12]}],"pathway":[{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0,1,2,7,8,12]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[16,17,18]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[2,8,21]}],"complexes":["Mediator complex"],"partners":["VP16","HNF4A","RARA","ETV4","ETV5","ATF6A","TP53","CREBBP"],"other_free_text":[]},"mechanistic_narrative":"MED25 is a subunit of the Mediator coactivator complex that serves as a principal docking platform for intrinsically disordered acidic transactivation domains of diverse transcription factors, thereby coupling activator-specific signals to RNA polymerase II recruitment at select promoters. Its central ACID domain adopts a seven-stranded β-barrel fold that presents two distinct binding faces: one shared by VP16 H1, ERM/ETV5, and p53 TAD2, and another engaged by VP16 H2, with cellular activators such as ATF6α, PEA3-group ETS factors, HNF4α, and nuclear receptors (RARα, ERα) each contacting the ACID or C-terminal LXXLL motif to recruit Mediator to their target genes [PMID:21378965, PMID:26130716, PMID:30360415, PMID:23531547, PMID:21135126, PMID:17641689]. MED25 occupancy promotes a permissive chromatin state by facilitating CREBBP-mediated H3K27 acetylation and opposing PRC2-dependent H3K27 trimethylation at target loci [PMID:25391650]. RSV NS1 protein exploits MED25 by straddling both ACID faces simultaneously with nanomolar affinity, competing with cellular activators to suppress antiviral gene expression and promote viral replication [PMID:40128225, PMID:40920851]."},"prefetch_data":{"uniprot":{"accession":"Q71SY5","full_name":"Mediator of RNA polymerase II transcription subunit 25","aliases":["Activator interaction domain-containing protein 1","Activator-recruited cofactor 92 kDa component","ARC92","Mediator complex subunit 25","p78"],"length_aa":747,"mass_kda":78.2,"function":"Component of the Mediator complex, a coactivator involved in the regulated transcription of nearly all RNA polymerase II-dependent genes. Mediator functions as a bridge to convey information from gene-specific regulatory proteins to the basal RNA polymerase II transcription machinery. Mediator is recruited to promoters by direct interactions with regulatory proteins and serves as a scaffold for the assembly of a functional preinitiation complex with RNA polymerase II and the general transcription factors. Required for RARA/RXRA-mediated transcription","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q71SY5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MED25","classification":"Not Classified","n_dependent_lines":279,"n_total_lines":1208,"dependency_fraction":0.23096026490066227},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000104973","cell_line_id":"CID000245","localizations":[{"compartment":"chromatin","grade":3},{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"MED10","stoichiometry":10.0},{"gene":"MED11","stoichiometry":10.0},{"gene":"MED14","stoichiometry":10.0},{"gene":"MED19","stoichiometry":10.0},{"gene":"H1F0","stoichiometry":10.0},{"gene":"MED24","stoichiometry":10.0},{"gene":"MED30","stoichiometry":10.0},{"gene":"MED16","stoichiometry":10.0},{"gene":"MED31","stoichiometry":10.0},{"gene":"MED27","stoichiometry":10.0}],"url":"https://opencell.sf.czbiohub.org/target/CID000245","total_profiled":1310},"omim":[{"mim_id":"616449","title":"BASEL-VANAGAITE-SMIRIN-YOSEF SYNDROME; BVSYS","url":"https://www.omim.org/entry/616449"},{"mim_id":"615095","title":"MICROCEPHALY 10, PRIMARY, AUTOSOMAL RECESSIVE; MCPH10","url":"https://www.omim.org/entry/615095"},{"mim_id":"610827","title":"ZINC FINGER PROTEIN 335; ZNF335","url":"https://www.omim.org/entry/610827"},{"mim_id":"610197","title":"MEDIATOR COMPLEX SUBUNIT 25; MED25","url":"https://www.omim.org/entry/610197"},{"mim_id":"609878","title":"MEDIATOR COMPLEX SUBUNIT 9; MED9","url":"https://www.omim.org/entry/609878"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MED25"},"hgnc":{"alias_symbol":["ARC92","ACID1","TCBAP0758","DKFZp434K0512"],"prev_symbol":[]},"alphafold":{"accession":"Q9NWA0","domains":[{"cath_id":"4.10.860","chopping":"57-104","consensus_level":"medium","plddt":91.4763,"start":57,"end":104},{"cath_id":"1.20.5","chopping":"109-140","consensus_level":"medium","plddt":91.7366,"start":109,"end":140}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NWA0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NWA0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NWA0-F1-predicted_aligned_error_v6.png","plddt_mean":75.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MED25","jax_strain_url":"https://www.jax.org/strain/search?query=MED25"},"sequence":{"accession":"Q9NWA0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NWA0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NWA0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NWA0"}},"corpus_meta":[{"pmid":"22822206","id":"PMC_22822206","title":"The Arabidopsis mediator subunit MED25 differentially regulates jasmonate and abscisic acid signaling through interacting with the MYC2 and ABI5 transcription factors.","date":"2012","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/22822206","citation_count":309,"is_preprint":false},{"pmid":"17560376","id":"PMC_17560376","title":"Purification of a plant mediator from Arabidopsis thaliana identifies PFT1 as the Med25 subunit.","date":"2007","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/17560376","citation_count":235,"is_preprint":false},{"pmid":"28973940","id":"PMC_28973940","title":"Mediator subunit MED25 links the jasmonate receptor to transcriptionally active chromatin.","date":"2017","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/28973940","citation_count":153,"is_preprint":false},{"pmid":"21343311","id":"PMC_21343311","title":"A high-throughput screening system for Arabidopsis transcription factors and its application to Med25-dependent transcriptional regulation.","date":"2011","source":"Molecular plant","url":"https://pubmed.ncbi.nlm.nih.gov/21343311","citation_count":137,"is_preprint":false},{"pmid":"21536906","id":"PMC_21536906","title":"The Arabidopsis thaliana Med25 mediator subunit integrates environmental cues to control plant development.","date":"2011","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/21536906","citation_count":116,"is_preprint":false},{"pmid":"24456400","id":"PMC_24456400","title":"The Arabidopsis Mediator subunit MED16 regulates iron homeostasis by associating with EIN3/EIL1 through subunit MED25.","date":"2014","source":"The Plant journal : for cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/24456400","citation_count":112,"is_preprint":false},{"pmid":"14983011","id":"PMC_14983011","title":"The activator-recruited cofactor/Mediator coactivator subunit ARC92 is a functionally important target of the VP16 transcriptional activator.","date":"2004","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/14983011","citation_count":110,"is_preprint":false},{"pmid":"31182849","id":"PMC_31182849","title":"MED25 connects enhancer-promoter looping and MYC2-dependent activation of jasmonate signalling.","date":"2019","source":"Nature plants","url":"https://pubmed.ncbi.nlm.nih.gov/31182849","citation_count":99,"is_preprint":false},{"pmid":"21378965","id":"PMC_21378965","title":"Structure and VP16 binding of the Mediator Med25 activator interaction domain.","date":"2011","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21378965","citation_count":95,"is_preprint":false},{"pmid":"21985558","id":"PMC_21985558","title":"PFT1, the MED25 subunit of the plant Mediator complex, promotes flowering through CONSTANS dependent and independent mechanisms in Arabidopsis.","date":"2011","source":"The Plant journal : for cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21985558","citation_count":94,"is_preprint":false},{"pmid":"17641689","id":"PMC_17641689","title":"MED25 is distinct from TRAP220/MED1 in cooperating with CBP for retinoid receptor activation.","date":"2007","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/17641689","citation_count":75,"is_preprint":false},{"pmid":"31852773","id":"PMC_31852773","title":"Mediator Subunit MED25 Couples Alternative Splicing of JAZ Genes with Fine-Tuning of Jasmonate Signaling.","date":"2019","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/31852773","citation_count":71,"is_preprint":false},{"pmid":"32777679","id":"PMC_32777679","title":"Mediator subunit MED25: at the nexus of jasmonate signaling.","date":"2020","source":"Current opinion in plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/32777679","citation_count":70,"is_preprint":false},{"pmid":"31320481","id":"PMC_31320481","title":"LEUNIG_HOMOLOG Mediates MYC2-Dependent Transcriptional Activation in Cooperation with the Coactivators HAC1 and MED25.","date":"2019","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/31320481","citation_count":63,"is_preprint":false},{"pmid":"21062487","id":"PMC_21062487","title":"Autotaxin and lysophosphatidic acid1 receptor-mediated demyelination of dorsal root fibers by sciatic nerve injury and intrathecal lysophosphatidylcholine.","date":"2010","source":"Molecular pain","url":"https://pubmed.ncbi.nlm.nih.gov/21062487","citation_count":63,"is_preprint":false},{"pmid":"19290556","id":"PMC_19290556","title":"Identification of the variant Ala335Val of MED25 as responsible for CMT2B2: molecular data, functional studies of the SH3 recognition motif and correlation between wild-type MED25 and PMP22 RNA levels in CMT1A animal models.","date":"2009","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/19290556","citation_count":58,"is_preprint":false},{"pmid":"21135126","id":"PMC_21135126","title":"Med25 is required for RNA polymerase II recruitment to specific promoters, thus regulating xenobiotic and lipid metabolism in human liver.","date":"2010","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21135126","citation_count":55,"is_preprint":false},{"pmid":"9856582","id":"PMC_9856582","title":"Plasma copolymer surfaces of acrylic acid/1,7 octadiene: surface characterisation and the attachment of ROS 17/2.8 osteoblast-like cells.","date":"1998","source":"Biomaterials","url":"https://pubmed.ncbi.nlm.nih.gov/9856582","citation_count":54,"is_preprint":false},{"pmid":"22992513","id":"PMC_22992513","title":"Proteasome-mediated turnover of Arabidopsis MED25 is coupled to the activation of FLOWERING LOCUS T transcription.","date":"2012","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/22992513","citation_count":44,"is_preprint":false},{"pmid":"36471914","id":"PMC_36471914","title":"Tomato MED25 regulates fruit ripening by interacting with EIN3-like transcription factors.","date":"2023","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/36471914","citation_count":42,"is_preprint":false},{"pmid":"18781168","id":"PMC_18781168","title":"A paramutation phenomenon is involved in the genetics of maize low phytic acid1-241 (lpa1-241) trait.","date":"2008","source":"Heredity","url":"https://pubmed.ncbi.nlm.nih.gov/18781168","citation_count":40,"is_preprint":false},{"pmid":"7626628","id":"PMC_7626628","title":"Contributions of individual kringle domains toward maintenance of the chloride-induced tight conformation of human glutamic acid-1 plasminogen.","date":"1995","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7626628","citation_count":37,"is_preprint":false},{"pmid":"32938743","id":"PMC_32938743","title":"Mediator Subunit MED25 Physically Interacts with PHYTOCHROME INTERACTING FACTOR4 to Regulate Shade-Induced Hypocotyl Elongation in Tomato.","date":"2020","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/32938743","citation_count":36,"is_preprint":false},{"pmid":"28728983","id":"PMC_28728983","title":"ETV4 and AP1 Transcription Factors Form Multivalent Interactions with three Sites on the MED25 Activator-Interacting Domain.","date":"2017","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/28728983","citation_count":34,"is_preprint":false},{"pmid":"33369119","id":"PMC_33369119","title":"Mediator tail module subunits MED16 and MED25 differentially regulate abscisic acid signaling in Arabidopsis.","date":"2021","source":"Journal of integrative plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/33369119","citation_count":32,"is_preprint":false},{"pmid":"20974256","id":"PMC_20974256","title":"NMR structure of the human Mediator MED25 ACID domain.","date":"2010","source":"Journal of structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/20974256","citation_count":31,"is_preprint":false},{"pmid":"31125430","id":"PMC_31125430","title":"Regulation of flowering time by SPL10/MED25 module in Arabidopsis.","date":"2019","source":"The New phytologist","url":"https://pubmed.ncbi.nlm.nih.gov/31125430","citation_count":31,"is_preprint":false},{"pmid":"10487890","id":"PMC_10487890","title":"Comparison of proliferation and growth of human keratinocytes on plasma copolymers of acrylic acid/1,7-octadiene and self-assembled monolayers.","date":"1999","source":"Journal of biomedical materials research","url":"https://pubmed.ncbi.nlm.nih.gov/10487890","citation_count":31,"is_preprint":false},{"pmid":"22239106","id":"PMC_22239106","title":"Protective role of D-saccharic acid-1,4-lactone in alloxan induced oxidative stress in the spleen tissue of diabetic rats is mediated by suppressing mitochondria dependent apoptotic pathway.","date":"2012","source":"Free radical research","url":"https://pubmed.ncbi.nlm.nih.gov/22239106","citation_count":30,"is_preprint":false},{"pmid":"23864652","id":"PMC_23864652","title":"Role for human mediator subunit MED25 in recruitment of mediator to promoters by endoplasmic reticulum stress-responsive transcription factor ATF6α.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23864652","citation_count":29,"is_preprint":false},{"pmid":"23531547","id":"PMC_23531547","title":"The Mediator complex subunit MED25 is targeted by the N-terminal transactivation domain of the PEA3 group members.","date":"2013","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/23531547","citation_count":28,"is_preprint":false},{"pmid":"25527630","id":"PMC_25527630","title":"Homozygous missense mutation in MED25 segregates with syndromic intellectual disability in a large consanguineous family.","date":"2014","source":"Journal of medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25527630","citation_count":28,"is_preprint":false},{"pmid":"30039206","id":"PMC_30039206","title":"The polynucleotide kinase 3'-phosphatase gene (PNKP) is involved in Charcot-Marie-Tooth disease (CMT2B2) previously related to MED25.","date":"2018","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/30039206","citation_count":28,"is_preprint":false},{"pmid":"21982801","id":"PMC_21982801","title":"D-saccharic acid-1,4-lactone ameliorates alloxan-induced diabetes mellitus and oxidative stress in rats through inhibiting pancreatic β-cells from apoptosis via mitochondrial dependent pathway.","date":"2011","source":"Toxicology and applied pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/21982801","citation_count":28,"is_preprint":false},{"pmid":"25792360","id":"PMC_25792360","title":"Homozygous MED25 mutation implicated in eye-intellectual disability syndrome.","date":"2015","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25792360","citation_count":26,"is_preprint":false},{"pmid":"22447446","id":"PMC_22447446","title":"Interactions between DNA, transcriptional regulator Dreb2a and the Med25 mediator subunit from Arabidopsis thaliana involve conformational changes.","date":"2012","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/22447446","citation_count":26,"is_preprint":false},{"pmid":"37606175","id":"PMC_37606175","title":"The PIF1/PIF3-MED25-HDA19 transcriptional repression complex regulates phytochrome signaling in Arabidopsis.","date":"2023","source":"The New phytologist","url":"https://pubmed.ncbi.nlm.nih.gov/37606175","citation_count":24,"is_preprint":false},{"pmid":"24874105","id":"PMC_24874105","title":"Interaction studies of the human and Arabidopsis thaliana Med25-ACID proteins with the herpes simplex virus VP16- and plant-specific Dreb2a transcription factors.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24874105","citation_count":24,"is_preprint":false},{"pmid":"21110951","id":"PMC_21110951","title":"PTOV1 antagonizes MED25 in RAR transcriptional activation.","date":"2010","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/21110951","citation_count":23,"is_preprint":false},{"pmid":"27082727","id":"PMC_27082727","title":"Lysophosphatidic Acid Signaling through the Lysophosphatidic Acid-1 Receptor Is Required for Alveolarization.","date":"2016","source":"American journal of respiratory cell and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/27082727","citation_count":23,"is_preprint":false},{"pmid":"36537119","id":"PMC_36537119","title":"The Mediator complex subunit MED25 interacts with HDA9 and PIF4 to regulate thermomorphogenesis.","date":"2023","source":"Plant physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36537119","citation_count":22,"is_preprint":false},{"pmid":"20191364","id":"PMC_20191364","title":"The low phytic acid1-241 (lpa1-241) maize mutation alters the accumulation of anthocyanin pigment in the kernel.","date":"2010","source":"Planta","url":"https://pubmed.ncbi.nlm.nih.gov/20191364","citation_count":20,"is_preprint":false},{"pmid":"26130716","id":"PMC_26130716","title":"Characterization of ERM transactivation domain binding to the ACID/PTOV domain of the Mediator subunit MED25.","date":"2015","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/26130716","citation_count":20,"is_preprint":false},{"pmid":"14640757","id":"PMC_14640757","title":"Direct electrochemical detection of oligonucleotide hybridization on poly(5-hydroxy-1,4-naphthoquinone- co-5-hydroxy-3-thioacetic acid-1,4-naphthoquinone) film.","date":"2003","source":"Analytical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14640757","citation_count":20,"is_preprint":false},{"pmid":"22563127","id":"PMC_22563127","title":"Study of low phytic acid1-7 (lpa1-7), a new ZmMRP4 mutation in maize.","date":"2012","source":"The Journal of heredity","url":"https://pubmed.ncbi.nlm.nih.gov/22563127","citation_count":19,"is_preprint":false},{"pmid":"21785987","id":"PMC_21785987","title":"Solution NMR structure of MED25(391-543) comprising the activator-interacting domain (ACID) of human mediator subunit 25.","date":"2011","source":"Journal of structural and functional genomics","url":"https://pubmed.ncbi.nlm.nih.gov/21785987","citation_count":17,"is_preprint":false},{"pmid":"26341899","id":"PMC_26341899","title":"The Mediator complex subunits MED25/PFT1 and MED8 are required for transcriptional responses to changes in cell wall arabinose composition and glucose treatment in Arabidopsis thaliana.","date":"2015","source":"BMC plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/26341899","citation_count":17,"is_preprint":false},{"pmid":"35212455","id":"PMC_35212455","title":"Mediator complex subunit MED25 physically interacts with DST to regulate spikelet number in rice.","date":"2022","source":"Journal of integrative plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/35212455","citation_count":16,"is_preprint":false},{"pmid":"15046766","id":"PMC_15046766","title":"Reagentless amperometric detection of l-lactate on an enzyme-modified conducting copolymer poly(5-hydroxy-1,4-naphthoquinone-co-5-hydroxy-3-thioacetic acid-1,4-naphthoquinone).","date":"2004","source":"Biosensors & bioelectronics","url":"https://pubmed.ncbi.nlm.nih.gov/15046766","citation_count":16,"is_preprint":false},{"pmid":"36102648","id":"PMC_36102648","title":"An Unexpected Encounter: Respiratory Syncytial Virus Nonstructural Protein 1 Interacts with Mediator Subunit MED25.","date":"2022","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/36102648","citation_count":15,"is_preprint":false},{"pmid":"21538728","id":"PMC_21538728","title":"Prophylactic role of D-Saccharic acid-1,4-lactone in tertiary butyl hydroperoxide induced cytotoxicity and cell death of murine hepatocytes via mitochondria-dependent pathways.","date":"2011","source":"Journal of biochemical and molecular toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/21538728","citation_count":15,"is_preprint":false},{"pmid":"28556372","id":"PMC_28556372","title":"Plant-plant interactions influence developmental phase transitions, grain productivity and root system architecture in Arabidopsis via auxin and PFT1/MED25 signalling.","date":"2017","source":"Plant, cell & environment","url":"https://pubmed.ncbi.nlm.nih.gov/28556372","citation_count":15,"is_preprint":false},{"pmid":"25391650","id":"PMC_25391650","title":"Epigenetic modification of histone 3 lysine 27: mediator subunit MED25 is required for the dissociation of polycomb repressive complex 2 from the promoter of cytochrome P450 2C9.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25391650","citation_count":15,"is_preprint":false},{"pmid":"20015711","id":"PMC_20015711","title":"Determination of D-saccharic acid-1,4-lactone from brewed kombucha broth by high-performance capillary electrophoresis.","date":"2009","source":"Journal of chromatography. B, Analytical technologies in the biomedical and life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/20015711","citation_count":14,"is_preprint":false},{"pmid":"22952853","id":"PMC_22952853","title":"MED25 is a mediator component of HNF4α-driven transcription leading to insulin secretion in pancreatic beta-cells.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22952853","citation_count":14,"is_preprint":false},{"pmid":"31986205","id":"PMC_31986205","title":"Nitrous oxide emissions from the urine of beef cattle as regulated by dietary crude protein and gallic acid1.","date":"2018","source":"Journal of animal science","url":"https://pubmed.ncbi.nlm.nih.gov/31986205","citation_count":13,"is_preprint":false},{"pmid":"30360415","id":"PMC_30360415","title":"Structural Basis for the Interaction between p53 Transactivation Domain and the Mediator Subunit MED25.","date":"2018","source":"Molecules (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/30360415","citation_count":13,"is_preprint":false},{"pmid":"23138840","id":"PMC_23138840","title":"The prophylactic role of D-saccharic acid-1,4-lactone against hyperglycemia-induced hepatic apoptosis via inhibition of both extrinsic and intrinsic pathways in diabetic rats.","date":"2013","source":"Food & function","url":"https://pubmed.ncbi.nlm.nih.gov/23138840","citation_count":12,"is_preprint":false},{"pmid":"30800049","id":"PMC_30800049","title":"COQ8A and MED25 Mutations in a Child with Intellectual Disability, Microcephaly, Seizures, and Spastic Ataxia: Synergistic Effect of Digenic Variants?","date":"2018","source":"Molecular syndromology","url":"https://pubmed.ncbi.nlm.nih.gov/30800049","citation_count":11,"is_preprint":false},{"pmid":"34929168","id":"PMC_34929168","title":"Protein-protein interactions between jasmonate-related master regulator MYC and transcriptional mediator MED25 depend on a short binding domain.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34929168","citation_count":11,"is_preprint":false},{"pmid":"27273787","id":"PMC_27273787","title":"Upregulation of human CYP2C9 expression by Bisphenol A via estrogen receptor alpha (ERα) and Med25.","date":"2016","source":"Environmental toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/27273787","citation_count":11,"is_preprint":false},{"pmid":"35907573","id":"PMC_35907573","title":"Respiratory Syncytial Virus NS1 Protein Targets the Transactivator Binding Domain of MED25.","date":"2022","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/35907573","citation_count":10,"is_preprint":false},{"pmid":"39824838","id":"PMC_39824838","title":"A multiprotein regulatory module, MED16-MBR1&2, controls MED25 homeostasis during jasmonate signaling.","date":"2025","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/39824838","citation_count":9,"is_preprint":false},{"pmid":"39739362","id":"PMC_39739362","title":"The AMS/DYT1-MYB module interacts with the MED25-MYC-MYB complexes to inhibit jasmonate-regulated floral defense in Arabidopsis.","date":"2024","source":"Journal of integrative plant biology","url":"https://pubmed.ncbi.nlm.nih.gov/39739362","citation_count":7,"is_preprint":false},{"pmid":"24960263","id":"PMC_24960263","title":"Med25 is required for estrogen receptor alpha (ERα)-mediated regulation of human CYP2C9 expression.","date":"2014","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/24960263","citation_count":7,"is_preprint":false},{"pmid":"32194589","id":"PMC_32194589","title":"Suppression of Arabidopsis Mediator Subunit-Encoding MED18 Confers Broad Resistance Against DNA and RNA Viruses While MED25 Is Required for Virus Defense.","date":"2020","source":"Frontiers in plant science","url":"https://pubmed.ncbi.nlm.nih.gov/32194589","citation_count":7,"is_preprint":false},{"pmid":"16966804","id":"PMC_16966804","title":"Solvent sublation and spectrometric determination of iron(II) and total iron using 3-(2-pyridyl)-5,6-bis(4-phenylsulfonic acid)-1,2,4-triazine and tetrabutylammonium bromide.","date":"2006","source":"Analytical sciences : the international journal of the Japan Society for Analytical Chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16966804","citation_count":7,"is_preprint":false},{"pmid":"35876657","id":"PMC_35876657","title":"Coordinately regulated transcription factors EIN3/EIL1 and MYCs in ethylene and jasmonate signaling interact with the same domain of MED25.","date":"2022","source":"Bioscience, biotechnology, and biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35876657","citation_count":5,"is_preprint":false},{"pmid":"37358186","id":"PMC_37358186","title":"Cooperative Lewis Acid-1,2,3-Triazolium-Aryloxide Catalysis: Pyrazolone Addition to Nitroolefins as Entry to Diaminoamides.","date":"2023","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/37358186","citation_count":5,"is_preprint":false},{"pmid":"28889249","id":"PMC_28889249","title":"Papillae formation on trichome cell walls requires the function of the mediator complex subunit Med25.","date":"2017","source":"Plant molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/28889249","citation_count":4,"is_preprint":false},{"pmid":"30410084","id":"PMC_30410084","title":"Origin and age of the causative mutations in KLC2, IMPA1, MED25 and WNT7A unravelled through Brazilian admixed populations.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/30410084","citation_count":4,"is_preprint":false},{"pmid":"32816121","id":"PMC_32816121","title":"Improving the phenotype description of Basel-Vanagaite-Smirin-Yosef syndrome, MED25-related: polymicrogyria as a distinctive neuroradiological finding.","date":"2020","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/32816121","citation_count":4,"is_preprint":false},{"pmid":"27314626","id":"PMC_27314626","title":"Design, semisynthesis and cytotoxic activity of novel ester derivatives of betulinic acid-1,2,4 oxadiazoles.","date":"2016","source":"Journal of Asian natural products research","url":"https://pubmed.ncbi.nlm.nih.gov/27314626","citation_count":4,"is_preprint":false},{"pmid":"37047128","id":"PMC_37047128","title":"Med25 Limits Master Regulators That Govern Adipogenesis.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/37047128","citation_count":4,"is_preprint":false},{"pmid":"25850761","id":"PMC_25850761","title":"Polymorphisms in the retinoic acid-1 like-receptor family of genes and their association with clinical outcome of dengue virus infection.","date":"2015","source":"Archives of virology","url":"https://pubmed.ncbi.nlm.nih.gov/25850761","citation_count":4,"is_preprint":false},{"pmid":"38847958","id":"PMC_38847958","title":"Genome-wide identification of the MED25 BINDING RING-H2 PROTEIN gene family in foxtail millet (Setaria italica L.) and the role of SiMBR2 in resistance to abiotic stress in Arabidopsis.","date":"2024","source":"Planta","url":"https://pubmed.ncbi.nlm.nih.gov/38847958","citation_count":3,"is_preprint":false},{"pmid":"38389893","id":"PMC_38389893","title":"Design, synthesis, and antiproliferative evaluation of novel dehydroabietic acid-1,2,3-triazole-oxazolidinone hybrids.","date":"2024","source":"RSC medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/38389893","citation_count":2,"is_preprint":false},{"pmid":"40128225","id":"PMC_40128225","title":"Molecular basis for human respiratory syncytial virus transcriptional regulator NS1 interactions with MED25.","date":"2025","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/40128225","citation_count":2,"is_preprint":false},{"pmid":"38527936","id":"PMC_38527936","title":"A Lipopeptidomimetic of Transcriptional Activation Domains Selectively Disrupts the Coactivator Med25 Protein-Protein Interactions.","date":"2024","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/38527936","citation_count":2,"is_preprint":false},{"pmid":"32825095","id":"PMC_32825095","title":"Multivalent and Bidirectional Binding of Transcriptional Transactivation Domains to the MED25 Coactivator.","date":"2020","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/32825095","citation_count":2,"is_preprint":false},{"pmid":"40545710","id":"PMC_40545710","title":"TurboID-Based Proximity Labelling Identifies MED25 as an Activator of GoPGF-Mediated Gland Formation and Gossypol Biosynthesis in Cotton.","date":"2025","source":"Plant, cell & environment","url":"https://pubmed.ncbi.nlm.nih.gov/40545710","citation_count":1,"is_preprint":false},{"pmid":"40920851","id":"PMC_40920851","title":"A dual interaction between RSV NS1 and MED25 ACID domain reshapes antiviral responses.","date":"2025","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/40920851","citation_count":0,"is_preprint":false},{"pmid":"41733769","id":"PMC_41733769","title":"Design, synthesis, and antibacterial efficacy of new methylene disalicylic acid/1,3,4-oxadiazole hybrids as dual inhibitors of DNA gyrase and topoisomerase IV.","date":"2026","source":"Molecular diversity","url":"https://pubmed.ncbi.nlm.nih.gov/41733769","citation_count":0,"is_preprint":false},{"pmid":"36993479","id":"PMC_36993479","title":"A lipopeptidomimetic of transcriptional activation domains selectively disrupts Med25 PPIs.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/36993479","citation_count":0,"is_preprint":false},{"pmid":"40909439","id":"PMC_40909439","title":"Unraveling the pathogenicity role of the novel compound heterozygous mutations of MED25 gene in a Chinese patient with BVSYS.","date":"2025","source":"Frontiers in genetics","url":"https://pubmed.ncbi.nlm.nih.gov/40909439","citation_count":0,"is_preprint":false},{"pmid":"38334938","id":"PMC_38334938","title":"Chemical shift assignments of the ACID domain of MED25, a subunit of the mediator complex in Arabidopsis thaliana.","date":"2024","source":"Biomolecular NMR assignments","url":"https://pubmed.ncbi.nlm.nih.gov/38334938","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.06.21.600057","title":"Genotype-environment association across precipitation regimens reveals the mechanism of plant adaptation to rainy environments","date":"2024-06-27","source":"bioRxiv","url":"https://doi.org/10.1101/2024.06.21.600057","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":41004,"output_tokens":5307,"usd":0.101308},"stage2":{"model":"claude-opus-4-6","input_tokens":9037,"output_tokens":3160,"usd":0.186278},"total_usd":0.287586,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2004,\n      \"finding\": \"Human MED25 (ARC92) was biochemically purified as a 92-kDa subunit of the ARC/Mediator coactivator complex that is specifically targeted by the activation domain of the VP16 transactivator; VP16 activation domain associates with ARC92 in vitro and in vivo, and siRNA-mediated knockdown of ARC92 selectively inhibits Gal4-VP16 gene activation.\",\n      \"method\": \"Affinity chromatography with VP16 activation domain, peptide microsequencing, in vitro and in vivo binding assays, siRNA knockdown with reporter gene assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — biochemical purification, in vitro and in vivo binding, functional siRNA validation; single rigorous study with multiple orthogonal methods\",\n      \"pmids\": [\"14983011\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Human MED25 associates with retinoic acid (RA)-bound RAR through its C-terminal LXXLL (NR box) motif, increases RAR/RXR-mediated transcription, and shows intrinsic transcriptional activity through its PTOV domain by direct association with CBP; MED25 is RA-dependently recruited to the RARβ2 promoter ahead of TRAP220, and MED25 knockdown selectively reduces RAR (but not TR) activity.\",\n      \"method\": \"Co-immunoprecipitation, ChIP, reporter assays with dominant-negative constructs, siRNA knockdown, domain mapping\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (Co-IP, ChIP, siRNA, reporter assays) in a single rigorous study\",\n      \"pmids\": [\"17641689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Human Med25 is required for HNF4α association with the Mediator complex and for RNA Pol II recruitment to a subset of HNF4α target gene promoters that selectively regulate drug and lipid metabolism; changes in endogenous Med25 levels alter the composition of the transcriptional complex and Pol II occupancy.\",\n      \"method\": \"Co-IP, ChIP, siRNA knockdown and overexpression with mRNA expression analysis\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, ChIP, gain- and loss-of-function with defined transcriptional phenotype\",\n      \"pmids\": [\"21135126\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"NMR structure of the MED25 activator interaction domain (ACID) reveals a seven-stranded β-barrel framed by three helices; VP16 subdomain H1 and H2 bind to opposite faces of ACID and cooperate during promoter-dependent activated transcription in vitro; the activator-binding ACID faces are functionally required and conserved among higher eukaryotes.\",\n      \"method\": \"NMR structure determination, mutagenesis, in vitro transcription assay\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR structure with mutagenesis and in vitro functional validation\",\n      \"pmids\": [\"21378965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"NMR structure of the human MED25 ACID domain reveals a closed β-barrel with seven strands and three α-helices, structurally similar to the SPOC domain-like superfamily; NMR chemical shift mapping shows VP16 H2 interacts with MED25 ACID through one face of the β-barrel defined by strands B4-B7-B6.\",\n      \"method\": \"NMR structure determination, NMR chemical shift mapping\",\n      \"journal\": \"Journal of structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR structure with interaction mapping\",\n      \"pmids\": [\"20974256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Solution NMR structure of human MED25(391-543) ACID domain shows a β-barrel architecture similar to Ku and SPOC domains; this domain interacts with the acidic transactivation domains of HSV-1 VP16 and VZV IE62.\",\n      \"method\": \"Solution NMR structure determination, backbone dynamics (15N-{1H} NOEs)\",\n      \"journal\": \"Journal of structural and functional genomics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR structure determination with functional context\",\n      \"pmids\": [\"21785987\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"MED25 plays a critical role in ATF6α-dependent recruitment of Mediator to ER stress response gene promoters; a specific MED25 domain serves as a docking site on Mediator for the ATF6α transcription activation domain.\",\n      \"method\": \"Co-IP, domain mapping, ChIP, in vitro binding\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and domain mapping with ChIP, single lab study\",\n      \"pmids\": [\"23864652\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The N-terminal acidic transactivation domain of PEA3 group ETS transcription factors (PEA3, ERM, ER81) directly contacts the ACID/PTOV domain of MED25; MED25 depletion disrupts ERM-Mediator association in vitro, reduces PEA3-driven MMP-1 expression, and Mediator recruitment; mutations preventing MED25-ERM binding strongly reduce transactivation.\",\n      \"method\": \"Co-IP, siRNA knockdown, reporter assays, domain mutagenesis, in vitro binding\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (Co-IP, siRNA, mutagenesis, reporter assay, ChIP) with defined functional phenotype\",\n      \"pmids\": [\"23531547\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MED25 is recruited to the CYP2C9 promoter through HNF4α; MED25 influences H3K27 acetylation status—overexpression enriches H3K27ac and CREBBP, while silencing leads to H3K27me3 and increased PRC2 occupancy at the promoter and on HNF4α itself; MED25 induces a permissive chromatin state (FAIRE) required for CYP2C9 transcription.\",\n      \"method\": \"ChIP, overexpression and siRNA silencing, FAIRE, in vitro transcription reconstitution\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP, FAIRE, in vitro reconstitution, gain and loss of function with epigenetic and transcriptional readouts\",\n      \"pmids\": [\"25391650\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MED25 interacts with HNF4α in pancreatic β-cells through the MED25 LXXLL motif in a ligand-independent manner; this interaction is required for full activation of HNF4α-mediated transcription and glucose-stimulated insulin secretion; MODY mutations at the LXXLL-binding pocket disrupt the interaction and impair insulin secretion via a loss-of-function mechanism.\",\n      \"method\": \"Yeast two-hybrid, Co-IP, in vitro binding, reporter assays, siRNA/shRNA knockdown, domain mutagenesis\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods with defined cellular phenotype (insulin secretion)\",\n      \"pmids\": [\"22952853\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The Y39C mutation in the MED25 von Willebrand factor type A (VWA) domain, which is responsible for MED25 recruitment into the Mediator complex, dramatically impairs MED25 interaction with the Mediator complex in mammalian cells, as shown by co-immunoprecipitation.\",\n      \"method\": \"Co-immunoprecipitation of mutant vs. wild-type MED25 with Mediator complex in mammalian cells\",\n      \"journal\": \"Human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP with disease-relevant mutation; single lab, single method\",\n      \"pmids\": [\"25792360\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"The ERM/ETV5 N-terminal acidic transactivation domain (TAD) folds upon binding to MED25 ACID, forming a fuzzy complex; two aromatic residues (F47 and W57) in ERM TAD mediate binding; mutation Q451 in the VP16 H1 binding pocket of MED25 affects ERM binding; ERM and VP16 H1 share a common binding interface on MED25.\",\n      \"method\": \"NMR spectroscopy, mutational analysis, competition experiments\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR with mutagenesis and competition experiments providing atomic-level detail\",\n      \"pmids\": [\"26130716\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ETV4 activation domain (AD) and DNA-binding domain (DBD) each independently interact with MED25, with the AD binding a single MED25 site and the DBD interacting with three MED25 sites, allowing simultaneous binding; MED25 stimulates ETV4 DNA binding in vitro by relieving autoinhibition; AP1 transcription factors JUN and FOS also contact MED25 at overlapping/distinct sites; genome-wide studies show ETV4 and MED25 co-occupy enhancers in prostate cancer cells.\",\n      \"method\": \"Binding assays (affinity measurements), in vitro DNA binding assays, ChIP-seq, functional enhancer assays\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — quantitative binding assays, in vitro functional assay, ChIP-seq with multiple orthogonal methods\",\n      \"pmids\": [\"28728983\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Med25 acts as a coactivator of ligand-activated ERα, interacting with ERα through its C-terminal LXXLL motif after BPA exposure, and is functionally involved in BPA-induced transcriptional regulation of CYP2C9 expression and enzyme activity.\",\n      \"method\": \"Reporter assays, Co-IP, ChIP, siRNA knockdown, enzyme activity assay\",\n      \"journal\": \"Environmental toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP, reporter assay, ChIP; single lab\",\n      \"pmids\": [\"27273787\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Med25 enhances ligand-dependent ERα-mediated transcriptional activation of CYP2C9 and interacts with activated ERα through its C-terminal LXXLL motif.\",\n      \"method\": \"Reporter assays, Co-IP, domain mapping\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — Co-IP and reporter assay; single lab, limited methods\",\n      \"pmids\": [\"24960263\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NMR spectroscopy and ITC characterization of the p53 transactivation domain (p53TAD) interaction with MED25 ACID shows that p53TAD2 sequence motif forms an amphipathic α-helix that binds an elongated hydrophobic groove of MED25 ACID; the same conserved mechanism is shared with ERM and VP16 TADs.\",\n      \"method\": \"NMR chemical shift perturbation, isothermal titration calorimetry, mutagenesis, structural modeling\",\n      \"journal\": \"Molecules (Basel, Switzerland)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — NMR, ITC, and mutagenesis with structural model\",\n      \"pmids\": [\"30360415\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RSV NS1 protein interacts with MED25 ACID domain through its C-terminal α3 helix (binding the H2 face of ACID, similar to other transcriptional activation domains such as VP16 and ATF6α), and the globular domain of NS1 also contributes; NS1 can compete with ATF6α TAD for MED25 binding; MED25 knockout enhances RSV replication, suggesting an antiviral role.\",\n      \"method\": \"Yeast two-hybrid, Co-IP (overexpression and endogenous during infection), NMR, in vitro binding, MED25 KO cell line with viral replication assay\",\n      \"journal\": \"Journal of virology; Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — NMR, Co-IP, KO functional assay, competition experiments; independently replicated across two papers\",\n      \"pmids\": [\"36102648\", \"35907573\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"X-ray crystal structure of the RSV NS1/MED25 ACID complex reveals NS1 straddles and binds two faces of MED25 ACID simultaneously—distinct from known viral activators; NS1 competes with transcription factor transactivation domains for MED25 binding; ChIP-seq and RNA-seq identify ATF3 as a transcription factor whose Mediator recruitment is affected by NS1/MED25 interaction.\",\n      \"method\": \"X-ray crystallography, NMR, ChIP-seq, RNA-seq, mutagenesis, in vitro binding\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with mutagenesis, NMR, and genome-wide functional validation\",\n      \"pmids\": [\"40128225\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NS1 α/β core domain binds to MED25 ACID cooperatively with NS1 α3 helix to achieve nanomolar affinity; single amino acid substitutions in NS1 α/β domain (notably E110A) significantly reduce NS1-MED25 affinity and attenuate RSV replication; in MED25-knockdown cells, rRSV replication is further attenuated, and the difference between WT and NS1 mutants is partially lost, indicating NS1-MED25 complex contributes to controlling antiviral responses.\",\n      \"method\": \"NMR, in vitro binding assays, mutagenesis, recombinant RSV replication assay, MED25 knockdown cells, ISG expression analysis\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — NMR, mutagenesis, KD cells with viral replication readout; multiple orthogonal methods\",\n      \"pmids\": [\"40920851\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"A short binding domain of MED25 (CMIDM) is responsible for interaction with MYC3 (plant ortholog context, but mechanistically relevant to MED25 function); bipartite interaction is critical for stable MYC3-MED25 complex formation; quantitative binding analysis reveals binding affinity order JAZJas < MED25CMIDM < JAZCMID, suggesting a mechanism for activation and negative feedback in jasmonate signaling.\",\n      \"method\": \"Biochemical binding assays (cell-free system), quantitative affinity measurements, domain mapping\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — quantitative in vitro binding with domain mapping; plant ortholog context\",\n      \"pmids\": [\"34929168\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"PTOV1 and MED25 reciprocally regulate RAR transcriptional activity through competitive binding to CBP; PTOV1 antagonizes MED25 by competing for CBP binding and opposing CBP recruitment to RA-responsive gene promoters, affecting RA sensitivity in cancer cells.\",\n      \"method\": \"Reporter assays, Co-IP, ChIP, overexpression/competition assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — Co-IP, ChIP, reporter assays; single lab\",\n      \"pmids\": [\"21110951\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Med25 silencing in 3T3-L1 preadipocytes enhances lipid accumulation and super-induces adipogenic master regulators C/EBPα and PPARγ, indicating that Med25 limits adipogenic potential by suppressing levels of these master regulators.\",\n      \"method\": \"siRNA-mediated silencing, adipogenesis induction, lipid staining, Western blot for C/EBPα and PPARγ\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — loss-of-function with defined cellular phenotype and molecular pathway; single lab\",\n      \"pmids\": [\"37047128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"A lipopeptidomimetic (LPPM-8) incorporating a medium-chain branched fatty acid binds the H2 face of the MED25 ACID activator interaction domain with ~4 μM Ki (>20-fold improved over unmodified peptide), stabilizes full-length MED25 in cells, and inhibits expression of genes regulated by Med25-activator protein-protein interactions in a triple-negative breast cancer cell model.\",\n      \"method\": \"In vitro binding assay (Ki determination), cellular proteome stabilization, gene expression analysis in cancer cells\",\n      \"journal\": \"Angewandte Chemie (International ed. in English)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — quantitative binding, cellular stabilization, and functional gene expression readout; single lab\",\n      \"pmids\": [\"38527936\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"Human MED25 is a structurally characterized subunit of the Mediator coactivator complex whose central ACID domain (a seven-stranded β-barrel) serves as a direct docking platform for the intrinsically disordered acidic transactivation domains of diverse transcription factors—including viral VP16, cellular p53, ETS factors (ETV4/ERM), ATF6α, nuclear receptors (RAR/RXR, HNF4α, ERα) via its C-terminal LXXLL motif, and viral NS1—thereby relaying activator-specific signals to RNA polymerase II; MED25 is required for Pol II recruitment to a selective subset of target gene promoters, influences epigenetic states (H3K27ac vs. H3K27me3) through modulation of HAT (CREBBP) and PRC2 occupancy, and is exploited by RSV NS1 protein, which binds two faces of the ACID domain to compete with cellular transcription factors and suppress antiviral gene expression.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MED25 is a subunit of the Mediator coactivator complex that serves as a principal docking platform for intrinsically disordered acidic transactivation domains of diverse transcription factors, thereby coupling activator-specific signals to RNA polymerase II recruitment at select promoters. Its central ACID domain adopts a seven-stranded β-barrel fold that presents two distinct binding faces: one shared by VP16 H1, ERM/ETV5, and p53 TAD2, and another engaged by VP16 H2, with cellular activators such as ATF6α, PEA3-group ETS factors, HNF4α, and nuclear receptors (RARα, ERα) each contacting the ACID or C-terminal LXXLL motif to recruit Mediator to their target genes [PMID:21378965, PMID:26130716, PMID:30360415, PMID:23531547, PMID:21135126, PMID:17641689]. MED25 occupancy promotes a permissive chromatin state by facilitating CREBBP-mediated H3K27 acetylation and opposing PRC2-dependent H3K27 trimethylation at target loci [PMID:25391650]. RSV NS1 protein exploits MED25 by straddling both ACID faces simultaneously with nanomolar affinity, competing with cellular activators to suppress antiviral gene expression and promote viral replication [PMID:40128225, PMID:40920851].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Establishing MED25 as a Mediator subunit and VP16 target resolved how the VP16 activation domain contacts the coactivator complex, identifying MED25 as the specific interface subunit required for VP16-driven transcription.\",\n      \"evidence\": \"Affinity purification with VP16 activation domain, peptide sequencing, in vivo binding, siRNA knockdown with reporter assay in human cells\",\n      \"pmids\": [\"14983011\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other transcription factors also dock on MED25 was unknown\", \"Structural basis of the VP16–MED25 interaction was unresolved\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Demonstrating that MED25 bridges ligand-bound RAR/RXR to Mediator via its LXXLL motif and recruits CBP via its PTOV domain established MED25 as a nuclear receptor coactivator with dual interaction surfaces.\",\n      \"evidence\": \"Co-IP, ChIP at RARβ2 promoter, reporter assays, siRNA knockdown, domain mapping in human cells\",\n      \"pmids\": [\"17641689\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How MED25 coordinates with other Mediator-NR contacts (e.g. MED1/TRAP220) was unclear\", \"The structural basis of the LXXLL–NR interaction was not resolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Three independent NMR structures of the MED25 ACID domain revealed a seven-stranded β-barrel architecture with distinct activator-binding faces, providing the first atomic framework for understanding how multiple transcription factors converge on MED25.\",\n      \"evidence\": \"NMR structure determination, chemical shift mapping of VP16 H2 binding surface\",\n      \"pmids\": [\"20974256\", \"21378965\", \"21785987\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How VP16 H1 and H2 cooperate on separate ACID faces during transcription was only partially resolved\", \"Whether cellular activators use the same or different binding surfaces was unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Showing that MED25 is required for HNF4α–Mediator association and Pol II recruitment to a selective subset of drug/lipid metabolism genes established that MED25 functions as a gene-selective coactivator rather than a general Mediator component.\",\n      \"evidence\": \"Co-IP, ChIP, siRNA knockdown and overexpression with mRNA analysis in human hepatic cells\",\n      \"pmids\": [\"21135126\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genome-wide scope of MED25-dependent genes was not defined\", \"How MED25 selectivity for specific HNF4α targets arises was unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identifying PEA3-group ETS factors and ATF6α as additional ACID-domain-dependent MED25 clients expanded the activator repertoire beyond VP16 and nuclear receptors, showing MED25 as a shared docking module for diverse acidic transactivation domains.\",\n      \"evidence\": \"Co-IP, siRNA, reporter assays, mutagenesis, ChIP, domain mapping for ERM/PEA3 and ATF6α\",\n      \"pmids\": [\"23531547\", \"23864652\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The structural basis for selectivity among different cellular activators at the ACID domain was not resolved\", \"Whether activator competition for MED25 occurs in vivo was untested\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrating that MED25 occupancy at target promoters toggles the H3K27ac/H3K27me3 balance by promoting CREBBP recruitment and opposing PRC2 revealed an epigenetic dimension to MED25 coactivator function beyond simple Pol II recruitment.\",\n      \"evidence\": \"ChIP for histone marks and CREBBP/PRC2, FAIRE, siRNA and overexpression, in vitro transcription reconstitution at CYP2C9 promoter\",\n      \"pmids\": [\"25391650\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the epigenetic switch is a direct MED25–CREBBP interaction effect or indirect was not fully dissected\", \"Genome-wide extent of MED25-dependent chromatin remodeling was not assessed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Linking MED25-HNF4α interaction via the LXXLL motif to glucose-stimulated insulin secretion in β-cells, and showing that MODY-associated HNF4α mutations disrupt this interface, connected MED25 function to a Mendelian disease mechanism.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, reporter assays, siRNA/shRNA knockdown, domain mutagenesis in pancreatic β-cells\",\n      \"pmids\": [\"22952853\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether MED25 variants themselves contribute to diabetes susceptibility was not tested\", \"In vivo validation in animal models was lacking\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"NMR characterization of the ERM TAD–MED25 ACID fuzzy complex, showing that ERM and VP16 H1 share a binding interface, unified the mechanism by which diverse disordered activation domains fold upon docking to a common ACID surface.\",\n      \"evidence\": \"NMR spectroscopy, mutational analysis, competition experiments\",\n      \"pmids\": [\"26130716\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full thermodynamic parameters for all activator–ACID interactions were incomplete\", \"Whether simultaneous multi-activator occupancy is possible was not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Discovering that MED25 stimulates ETV4 DNA binding by relieving autoinhibition, and that ETV4 and MED25 co-occupy enhancers genome-wide, revealed an unexpected allosteric function of MED25 beyond passive scaffold bridging.\",\n      \"evidence\": \"Quantitative binding assays, in vitro DNA binding, ChIP-seq in prostate cancer cells\",\n      \"pmids\": [\"28728983\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether MED25 relieves autoinhibition for other ETS or non-ETS factors was unknown\", \"Structural basis of the allosteric effect was not determined\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Structural characterization of p53 TAD2 binding to MED25 ACID confirmed that a conserved amphipathic helix mechanism is shared by VP16, ERM, and p53, establishing a unifying structural code for MED25-activator recognition.\",\n      \"evidence\": \"NMR chemical shift perturbation, ITC, mutagenesis\",\n      \"pmids\": [\"30360415\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequences of p53–MED25 interaction for p53 target gene transcription were not assessed in cells\", \"Relative in vivo affinities of competing activators remained unmeasured\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showing that RSV NS1 binds MED25 ACID via its α3 helix at the H2 face and competes with ATF6α, with MED25 knockout enhancing viral replication, established MED25 as a host antiviral factor exploited by a pathogen.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP (endogenous during infection), NMR, MED25 KO cells with viral replication assay\",\n      \"pmids\": [\"36102648\", \"35907573\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The full structural basis of the NS1–MED25 complex was not resolved\", \"Which specific antiviral genes are controlled through MED25 was unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Crystal structure of the NS1–MED25 ACID complex revealed NS1 straddles both ACID faces simultaneously—a binding mode distinct from any known cellular or viral activator—and ChIP-seq/RNA-seq identified ATF3 as a transcription factor whose Mediator recruitment is disrupted by NS1, defining the antiviral gene program controlled through MED25.\",\n      \"evidence\": \"X-ray crystallography, NMR, ChIP-seq, RNA-seq, mutagenesis, recombinant RSV replication in MED25-knockdown cells\",\n      \"pmids\": [\"40128225\", \"40920851\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether additional transcription factors beyond ATF3 and ATF6α are major in vivo targets of NS1-mediated displacement is not fully mapped\", \"Therapeutic targeting of the NS1–MED25 interface has not been validated in vivo\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A complete in vivo hierarchy of activator competition at the MED25 ACID domain, genome-wide definition of MED25-dependent versus MED25-independent Mediator function, and the structural basis of MED25 integration into the full Mediator complex remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No cryo-EM or crystal structure of MED25 within intact Mediator–Pol II PIC\", \"Genome-wide quantitative model of activator competition for MED25 in physiological contexts is lacking\", \"In vivo animal models with MED25 loss-of-function phenotyping are limited\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 1, 2, 7, 8, 9, 12]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 6, 7, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [1, 8, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0, 1, 2, 7, 8, 12]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [16, 17, 18]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [2, 8, 21]}\n    ],\n    \"complexes\": [\n      \"Mediator complex\"\n    ],\n    \"partners\": [\n      \"VP16\",\n      \"HNF4A\",\n      \"RARA\",\n      \"ETV4\",\n      \"ETV5\",\n      \"ATF6A\",\n      \"TP53\",\n      \"CREBBP\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}