{"gene":"MAP2K7","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":1997,"finding":"MKK7 (JNKK2) was cloned and identified as a novel MAPKK that is specific for the SAPK/JNK subgroup; unlike SEK1/MKK4, it does not activate p38. MKK7 directly phosphorylates and activates JNK/SAPK. It is activated by TNFα and environmental stresses, and immunochemical studies identified MKK7 as a major JNK-activating kinase in osmotically shocked cells. Overexpression of MKK7 enhanced AP-1-dependent transcription.","method":"Molecular cloning, immunoprecipitation, in vitro kinase assay, co-expression studies, reporter assay","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — original cloning paper with in vitro kinase assay, substrate specificity demonstrated, replicated across subsequent papers","pmids":["9384583"],"is_preprint":false},{"year":1997,"finding":"MKK7 was identified as a murine homolog of Drosophila Hemipterous; it functionally rescues hep mutant flies. In fibroblasts, MKK7 is activated by stress and by GTPase Rac1. MKK7 directly phosphorylates and activates JNK/SAPK.","method":"Molecular cloning, Drosophila complementation rescue, in vitro kinase assay, transfection in fibroblasts","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay plus in vivo complementation, replicated by multiple labs","pmids":["9312105"],"is_preprint":false},{"year":1997,"finding":"Human JNKK2 (MKK7) was cloned and shown to be a highly specific JNK kinase that does not activate p38 MAPK. Unlike JNKK1/MKK4, it is JNK-specific.","method":"Database search, cDNA cloning, co-expression kinase assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct cloning and biochemical characterization, replicated by multiple labs","pmids":["9372971"],"is_preprint":false},{"year":1997,"finding":"MKK7 (but not SEK1/MKK4) is activated by Fas signaling as the upstream activator of JNK/SAPK; MKK6 is the major p38 activator in this pathway. JNK/SAPK activation in Fas signaling does not require CPP32-like proteases.","method":"Immune complex kinase assay, peptide inhibitor pharmacology, immunoprecipitation","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — kinase assay with multiple inhibitor controls, single lab","pmids":["9362518"],"is_preprint":false},{"year":1997,"finding":"MKK7 is the activator of JNK/SAPK activated by IL-1 in rabbit liver; it is not activated by MKK4 antibodies. JNK/SAPK is the only MAPK activated by IL-1 in liver.","method":"S-Sepharose purification, immunoprecipitation with anti-MKK7 and anti-MKK4 antisera, kinase assay","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — immunoprecipitation and kinase assay, single lab, replicated conceptually by others","pmids":["9414114"],"is_preprint":false},{"year":1998,"finding":"Human MKK7 (47 kDa) specifically phosphorylates and activates JNK1 but fails to activate p38 MAPK in co-expression studies. MKK7 is activated in hematopoietic cells by IL-3, CD40 ligation, B-cell antigen receptor, FcR, heat, UV, anisomycin, hyperosmolarity, TNFα, and by constitutively active Ras, Rac, and Cdc42.","method":"Molecular cloning, co-expression, immunoprecipitation kinase assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — direct biochemical characterization with substrate specificity, replicated across labs","pmids":["9535930"],"is_preprint":false},{"year":1998,"finding":"MKK7 plays a major role in SAPK/JNK activation in T lymphocytes responding to TCR/co-stimulation. Dominant-negative MKK7 abrogated transcriptional activation of the distal NFAT response element in the IL-2 promoter. Both the MKK6-p38 and MKK7-JNK pathways are activated in a cyclosporin A-sensitive manner and contribute to IL-2 gene expression.","method":"Dominant-negative transfection, reporter assay, kinase assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dominant-negative and reporter assay, single lab, conceptually replicated","pmids":["9575191"],"is_preprint":false},{"year":1999,"finding":"The MKK7 gene encodes six isoforms (α1/2, β1/2, γ1/2) by alternative splicing with three different N-termini and two C-termini. The N-terminal extension (absent in MKK7α) binds directly to the MKK7 substrate JNK. MKK7α isoforms show lower basal activity but higher inducible fold-activation than β and γ isoforms. MKK7 is detected in both cytoplasmic and nuclear compartments, but nuclear localization is not required for JNK activation in vivo.","method":"Molecular cloning, yeast two-hybrid/direct binding, kinase assay, immunofluorescence","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — molecular cloning with direct binding and kinase assays, multiple orthogonal methods","pmids":["9891090"],"is_preprint":false},{"year":1999,"finding":"DLK (dual leucine zipper-bearing kinase) associates with, phosphorylates, and activates MKK7 in vitro and in vivo, but unlike MLK3 does not phosphorylate or activate MKK4. DLK and MKK7 co-localize in neurons and occupy similar subcellular compartments, distinct from MKK4.","method":"In vitro kinase assay, co-immunoprecipitation, immunocytochemistry, subcellular fractionation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — in vitro kinase assay, reciprocal co-IP, and subcellular fractionation with functional consequence","pmids":["10187804"],"is_preprint":false},{"year":1999,"finding":"MEKK3 (and MEKK2) directly activates MKK7 and MKK6 in vitro; immunoprecipitates of MEKK3 phosphorylate recombinant MKK7 in vitro. Coexpression of MKK7 with MEKK3 in COS-7 cells enhanced MKK7 autophosphorylation and its ability to activate JNK1.","method":"Co-expression, in vitro kinase assay with recombinant proteins, immunoprecipitation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with recombinant proteins plus cellular co-expression, single lab","pmids":["10347227"],"is_preprint":false},{"year":1999,"finding":"MST/MLK2 activates recombinant MKK7 more effectively than recombinant SEK1/MKK4 in vitro. The majority of MLK2-dependent JNK-activating activity co-fractionates with MKK7, not MKK4. The MLK2 kinase domain determines this substrate specificity.","method":"In vitro reconstitution kinase assay, column fractionation, Western blot","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay with deletion analysis, single lab","pmids":["9516438"],"is_preprint":false},{"year":1999,"finding":"A JNKK2-JNK1 fusion protein acts as a constitutively active Jun kinase; JNK1 is phosphorylated by JNKK2 at both Thr183 and Tyr185 in the fusion. The fusion protein is specific for the JNK pathway (does not activate p38 or ERK2) and is sufficient to stimulate c-Jun transcriptional activity. The fusion protein localizes predominantly to the nucleus.","method":"Fusion protein construction, immunoblotting with phospho-specific antibodies, in vitro kinase assay, reporter assay, immunofluorescence","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal methods including phospho-specific immunoblot, kinase assay, and transcriptional reporter","pmids":["10506143"],"is_preprint":false},{"year":1999,"finding":"G protein βγ subunit activates MKK7 by ~2-fold in a Rac-dependent manner; this is distinct from Gbγ-induced MKK4 activation which depends on Rho and Cdc42. MKK7 activation by Gbγ is not blocked by tyrosine kinase inhibitors PP2/PP1, unlike MKK4 activation.","method":"Transfection with kinase-deficient mutants, dominant-negative GTPases, kinase activity assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dominant-negative epistasis and kinase assay, single lab","pmids":["9890951"],"is_preprint":false},{"year":2000,"finding":"MKK4 shows a striking preference for phosphorylating Tyr185 of SAPK1/JNK isoforms, while MKK7 shows a striking preference for Thr183. Together, MKK4 and MKK7 produce a synergistic increase in JNK activity in vitro. MKK7β is several hundred-fold more efficient than MKK7α' in activating JNK isoforms.","method":"In vitro kinase assay, phosphoamino acid analysis, mass spectrometry","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — rigorous in vitro reconstitution with site-specific phosphorylation analysis across multiple JNK isoforms","pmids":["11062067"],"is_preprint":false},{"year":2000,"finding":"MKK7 monophosphorylates JNK3α1 at Thr183 (Thr residue) in vitro; both MKK4 and MKK7 are required for bisphosphorylation and maximal enzyme activity of JNK3α1. MKK4 alone shows no phosphorylation of JNK3α1 by mass spectrometry.","method":"In vitro kinase assay, mass spectral phosphorylation analysis, kinetic measurements","journal":"Biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — mass spectrometry-verified phosphorylation sites with kinetic characterization","pmids":["10715136"],"is_preprint":false},{"year":2001,"finding":"Genetic disruption of Mkk7 alone was sufficient to prevent JNK activation by proinflammatory cytokines, whereas simultaneous disruption of Mkk4 and Mkk7 was required to block JNK activation by environmental stress. MKK7 preferentially phosphorylates JNK on Thr and MKK4 on Tyr.","method":"Targeted gene disruption (knockout mice), JNK kinase assay, phospho-specific analysis","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knockout with biochemical readout, demonstrates non-redundant in vivo roles, replicated by multiple labs","pmids":["11390361"],"is_preprint":false},{"year":2001,"finding":"MKK7 is an essential and specific regulator of SAPK/JNK activation in hematopoietic cells. Loss of MKK7 in thymocytes, mature B cells, and mast cells causes hyperproliferation; SAPK/JNK activation was completely abolished in mkk7−/− mast cells despite normal MKK4 phosphorylation. MKK7 negatively regulates proliferation through a pathway involving p16INK4a; reexpression of p16INK4a abrogates the hyperproliferative response.","method":"Conditional gene targeting, kinase assay, flow cytometry, Western blotting, reexpression rescue","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout with mechanistic rescue experiment (p16 reexpression), replicated across multiple cell types","pmids":["11560992"],"is_preprint":false},{"year":2001,"finding":"PKCδ mediates ionizing radiation-induced JNK activation through MKK7 (not MKK4) in human thyroid cells. IR activates MKK7 but not MKK4; this was blocked by the PKCδ inhibitor rottlerin and by kinase-deficient MKK7, defining a PKCδ→MKK7→JNK→AP-1 cascade.","method":"Dominant-negative adenoviral expression, immune-complex kinase assay, pharmacological inhibitor","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — kinase assay with dominant-negative and pharmacological inhibitor, single lab","pmids":["11314034"],"is_preprint":false},{"year":2001,"finding":"SEK1 (MKK4) and MKK7 show synergistic SAPK/JNK activation in embryonic stem cells. MKK7 is responsible for Thr phosphorylation of JNK; without MKK7, Thr phosphorylation is lost but Tyr phosphorylation remains. MKK7α1 requires prior Tyr phosphorylation by SEK1 before it can phosphorylate JNK Thr; MKK7γ1 phosphorylates Thr independently; MKK7γ2 can phosphorylate both Thr and Tyr.","method":"Knockout ES cells, dual phosphorylation analysis, co-transfection with kinase-dead mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knockout combined with phospho-specific analyses and multiple isoform comparisons","pmids":["11418587"],"is_preprint":false},{"year":2002,"finding":"SKRP1, a novel dual-specificity phosphatase, interacts with MKK7 (co-precipitation in vitro and in vivo) and inactivates the JNK pathway by dephosphorylating JNK. SKRP1 does not bind directly to JNK but gains access to it through MKK7. SKRP1 does not interfere with MKK7-JNK co-precipitation.","method":"Co-immunoprecipitation, in vitro binding assay, in vitro phosphatase assay, overexpression","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — co-IP in vitro and in vivo, phosphatase assay, single lab","pmids":["11959861"],"is_preprint":false},{"year":2002,"finding":"ZAK MAP3K utilizes MKK7 (not MKK4) to activate JNK/SAPK; dominant-negative MKK7 (but not MKK4) attenuated ZAK-induced JNK activation. ZAK activity disrupts actin stress fibers and causes G2/M cell cycle arrest.","method":"Co-expression, dominant-negative kinase mutants, flow cytometry, Western blot","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — dominant-negative overexpression only, single lab, single method","pmids":["12220515"],"is_preprint":false},{"year":2003,"finding":"In fibroblast-like synoviocytes, JNK, MKK4, and MKK7 form a stable complex detectable by co-immunoprecipitation, and MKK4 co-precipitates with MKK7. The complex localizes in the cytoplasm by confocal microscopy; JNK migrates to the nucleus after IL-1 stimulation. The complex is functionally active and phosphorylates c-Jun after IL-1.","method":"Co-immunoprecipitation, confocal microscopy, in vitro kinase assay","journal":"Arthritis and rheumatism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP and confocal colocalization with functional kinase readout, single lab","pmids":["13130464"],"is_preprint":false},{"year":2003,"finding":"In mkk7−/− embryonic stem cells, Thr phosphorylation of JNK is lost while Tyr phosphorylation remains. In 293T cells, SEK1-induced Tyr phosphorylation of JNK1 is followed by additional Thr phosphorylation by MKK7. SEK1 (but not MKK7) binds to JNK1 in 293T cells, indicating sequential rather than independent phosphorylation.","method":"Knockout ES cells, phospho-specific immunoblot, co-immunoprecipitation, Thr-Pro-Phe JNK1 mutant","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — genetic knockout, co-IP, and mutant JNK analysis across two cell systems","pmids":["12624093"],"is_preprint":false},{"year":2004,"finding":"GADD45β mediates NF-κB suppression of JNK signaling by directly binding MKK7 and blocking its catalytic activity. Gadd45β binds MKK7 directly; peptides disrupting the GADD45β/MKK7 interaction hinder the ability of both GADD45β and NF-κB to suppress TNFα-induced cytotoxicity.","method":"Unbiased screen, direct binding assay, kinase activity assay, peptide disruption, cell viability assay","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — direct binding demonstrated, kinase inhibition assayed, functional rescue with disrupting peptides","pmids":["14743220"],"is_preprint":false},{"year":2004,"finding":"MKK7 couples stress signaling to G2/M cell cycle progression and cellular senescence. Genetic inactivation of MKK7 in MEFs leads to premature senescence and G2/M cell cycle arrest. The CDC2 kinase is a downstream target of the MKK7-JNK-c-Jun pathway; loss of c-Jun or JNK phosphorylation sites on c-Jun produces the same G2/M block.","method":"Gene knockout (MEFs), cell cycle analysis, epistasis with c-Jun mutants, CDC2 target identification","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout with defined cell cycle phenotype and pathway placement through c-Jun mutant epistasis","pmids":["15039780"],"is_preprint":false},{"year":2005,"finding":"MLK3 activates MKK7 in the MLK3→MKK7→JNK cascade in hippocampal CA1 neurons after cerebral ischemia; K252a (MLK3 inhibitor) blocked MKK7 and JNK3 activation and was neuroprotective.","method":"In vivo ischemia model, immunoprecipitation kinase assay, pharmacological inhibition","journal":"Neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo kinase assay with pharmacological inhibitor, single lab","pmids":["15680699"],"is_preprint":false},{"year":2006,"finding":"MKK7 contains three JNK-docking D-sites (D1, D2, D3) in its N-terminal domain that cooperatively promote binding to JNK1. Mutation of any single site reduces binding by 50–70%; mutation of all three reduces binding by 95%. Full-length MKK7 with D1/D2 mutations has reduced JNK1 kinase activation. D-site peptides from MKK7 inhibit JNK1 and JNK2 substrate phosphorylation selectively over ERK2.","method":"Site-directed mutagenesis, binding assay, JNK kinase assay, peptide inhibition","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — systematic mutagenesis of all three docking sites with quantitative binding and functional kinase readouts","pmids":["16533805"],"is_preprint":false},{"year":2006,"finding":"c-FLIPL directly interacts with MKK7 in a TNFα-dependent manner and inhibits the interactions of MKK7 with MEKK1, ASK1, and TAK1. This interaction selectively suppresses the prolonged phase of JNK activation and reduces ROS accumulation in NF-κB-deficient fibroblasts.","method":"Co-immunoprecipitation, Western blot, c-Flip knockout cells, overexpression","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP in stimulated cells and knockout fibroblasts, single lab, two orthogonal approaches","pmids":["17110930"],"is_preprint":false},{"year":2006,"finding":"In fibroblast-like synoviocytes, IL-1β-induced JNK phosphorylation and activity (measured by kinase assay), AP-1 binding, AP-1-driven transcription, and MMP-3 production are dependent on MKK7 but not MKK4. Anisomycin-induced JNK activation requires both MKK4 and MKK7.","method":"siRNA knockdown, in vitro kinase assay, EMSA, AP-1 reporter assay, ELISA","journal":"Arthritis and rheumatism","confidence":"High","confidence_rationale":"Tier 2 / Strong — siRNA knockdown with multiple orthogonal functional readouts, single lab","pmids":["16802349"],"is_preprint":false},{"year":2007,"finding":"GADD45β is a structured protein whose helices α3/α4 and loops 1/2 mediate association with MKK7. Helix α3 primarily mediates docking; loop 1 and α4-loop 2 engage the ATP-binding site of MKK7 and cause conformational changes that impede catalytic function.","method":"Structural prediction, mutagenesis, binding/inhibition assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 1-3 / Moderate — structural prediction with mutagenesis mapping of interaction, single lab; not a full crystal structure","pmids":["17485467"],"is_preprint":false},{"year":2008,"finding":"GADD45β forms homodimers in solution (via helices 1 and 5) and binds tightly to MKK7; the dimerization interface (helices 1 and 5) is distinct from the MKK7-binding region. The complex may form at least a tetrameric MKK7–GADD45β:GADD45β–MKK7 unit.","method":"Size exclusion chromatography, NMR, biophysical binding assays","journal":"Journal of molecular biology","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — NMR and biophysical characterization of complex, single lab","pmids":["18343408"],"is_preprint":false},{"year":2008,"finding":"The IL-1–MKK7–JNK–c-Jun signaling pathway induces EGR-1 transcription via an AP-1 site and three distal SREs. c-Jun is required for EGR-1 transcription through both elements; IL-1-inducible c-Jun recruitment to both AP-1 and SRE regions was confirmed by ChIP. EGR-1 in turn binds promoters of IL-6, IL-8, and CCL2.","method":"JNK-MKK7 fusion protein, genome-wide microarray, qPCR, reporter assay, chromatin immunoprecipitation, siRNA knockdown","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods including ChIP, microarray, reporter, and genetic KO cells","pmids":["18281687"],"is_preprint":false},{"year":2009,"finding":"Rac1/MKK7/JNK signaling upregulates Atg5 in response to oncogenic H-Ras, which is critically required for autophagic cell death. Inhibition of MKK7 (but not MKK4) attenuated H-Ras(V12)-induced JNK activation; Rac1 siRNA or dominant-negative Rac1 inhibited MKK7-JNK activation and Atg5 upregulation.","method":"siRNA knockdown, dominant-negative GTPase, immunoblotting, cell death assays","journal":"Carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA and dominant-negative epistasis defining pathway, single lab","pmids":["19783847"],"is_preprint":false},{"year":2010,"finding":"Filamin A (and B, C) binds to MKK7 (specifically to the N-terminal residues 31–60 present in MKK7β and MKK7γ but not MKK7α) and also binds MKK4, connecting them in close proximity. MKK7γ (but not MKK7α) co-localizes with actin stress fibers and Filamin A. In Filamin A-deficient cells, MKK7 activation is reduced and stress-induced synergistic JNK activation is impaired.","method":"Co-immunoprecipitation, Filamin A-deficient cell line, deletion mutant analysis, confocal microscopy, kinase assay","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP, loss-of-function cell line, domain mapping, colocalization, all orthogonal","pmids":["20156194"],"is_preprint":false},{"year":2011,"finding":"MKK7 acts as a tumor suppressor coupling oncogenic/genotoxic stress to p53 stability in vivo. Tissue-specific inactivation of MKK7 in KRas(G12D)-driven lung and NeuT-driven mammary tumors markedly accelerates tumor onset. Mechanistically, MKK7 acts through JNK1 and JNK2, and this pathway directly couples oncogenic stress to p53 stability, required for cell cycle arrest.","method":"Conditional knockout (tissue-specific), tumor model, epistasis with JNK1/JNK2 knockouts, p53 stability assays","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo conditional knockout in two independent cancer models with JNK epistasis analysis","pmids":["21317887"],"is_preprint":false},{"year":2011,"finding":"MKK7-deficient MEFs display premature senescence and G2/M arrest; MKK7 deletion in cardiomyocytes leads to heart failure under pressure overload with increased apoptosis (elevated p53, reduced MnSOD) and fibrosis (upregulated TGF-β signaling).","method":"Cardiomyocyte-specific conditional knockout, pressure overload surgery, cell death assays, Western blot","journal":"Journal of molecular and cellular cardiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional knockout with defined cardiac phenotype and mechanistic protein markers, single lab","pmids":["21284947"],"is_preprint":false},{"year":2011,"finding":"In mkk7−/− mast cells, expression of JunB and p16INK4a are reduced and cyclin D1 is upregulated; reexpression of p16INK4a abrogates the hyperproliferative response. MKK7-regulated JNK signaling thus maintains cell cycle brakes via the p16INK4a axis.","method":"Gene knockout, flow cytometry, Western blot, adenoviral reexpression rescue","journal":"The Journal of experimental medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockout with mechanistic rescue, single lab study but landmark paper","pmids":["11560992"],"is_preprint":false},{"year":2011,"finding":"Neuron-specific deletion of MKK7 impairs brain development: mice show enlarged ventricles, reduced striatum, absent axon tracts, and abnormal filamentous accumulations. MKK7 regulates axon elongation in a cell-autonomous manner and is required for contralateral axon projection by cortical neurons. JNK substrate phosphorylation (c-Jun, neurofilament heavy chain, MAP1B, doublecortin) is reduced in MKK7-deficient brain.","method":"Nestin-Cre conditional knockout, histology, electron microscopy, in utero electroporation, in vitro axon elongation assay, immunoblot of JNK substrates","journal":"The Journal of neuroscience","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout with cell-autonomous in vitro confirmation and multiple JNK substrate readouts","pmids":["22090513"],"is_preprint":false},{"year":2012,"finding":"MKK7 mRNA localizes to the growth cone where it can be translated. MKK7 protein is phosphorylated specifically in the neurite shaft and is part of a DLK–MKK7–JNK1 module that triggers MAP1b phosphorylation to regulate microtubule bundling and neurite elongation. This local translation mechanism positions JNK signaling to microtubule regulation while uncoupling it from nuclear/transcriptional functions.","method":"Genome-wide screen for neurite-enriched mRNAs, FISH/imaging for mRNA localization, live cell imaging, immunofluorescence, pharmacological and genetic manipulation","journal":"PLoS biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide screen, mRNA localization, kinase module identification with functional output, multiple orthogonal methods","pmids":["23226105"],"is_preprint":false},{"year":2012,"finding":"TIPRL (TOR signaling pathway regulator-like protein) binds to MKK7 and promotes the interaction between MKK7 and PP2Ac (protein phosphatase 2A catalytic subunit), resulting in dephosphorylation/inactivation of MKK7 and JNK, thereby conferring resistance to TRAIL-induced apoptosis in HCC cells.","method":"Co-immunoprecipitation, GST pulldown, siRNA knockdown, phospho-immunoblot, confocal microscopy","journal":"Gastroenterology","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP, pulldown, knockout cells, and mechanistic phosphatase interaction identified","pmids":["22841785"],"is_preprint":false},{"year":2013,"finding":"Neuregulin/ErbB2 signaling activates Rac1, which activates MKK7 and JNK to upregulate c-Jun and downregulate Krox20 during Schwann cell dedifferentiation after nerve injury. Rac inhibition blocked MKK7 activation and c-Jun induction in sciatic nerves after axotomy.","method":"Primary Schwann cell culture, pharmacological Rac inhibition, immunoblot, ErbB2 inhibitor, microarray, axotomy model","journal":"Glia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological and in vivo inhibitor studies defining signaling pathway, single lab","pmids":["23505039"],"is_preprint":false},{"year":2014,"finding":"GADD45β/MKK7 interaction was identified as a therapeutic target in multiple myeloma. DTP3, a D-tripeptide, disrupts the GADD45β/MKK7 complex, restores MKK7/JNK kinase activity, and kills MM cells selectively. DTP3 ablates myeloma xenografts in mice.","method":"Drug discovery/screening, co-immunoprecipitation disruption assay, JNK kinase assay, cell viability, xenograft mouse model","journal":"Cancer cell","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — direct complex disruption assay, kinase activity restoration, in vivo xenograft","pmids":["25314077"],"is_preprint":false},{"year":2014,"finding":"MAVS recruits MKK7 onto mitochondria via its 3D domain upon viral infection. MKK7 subsequently phosphorylates JNK2 (not JNK1) to activate virus-induced apoptosis. Mkk7−/− cells fail to initiate virus-induced apoptosis; MAVS-MKK7-JNK2 defines a novel antiviral apoptotic pathway.","method":"Knockout cell lines (Mkk7−/−, Jnk1−/−, Jnk2−/−), co-immunoprecipitation, mitochondrial fractionation, viral challenge, Jnk2−/− mice","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic knockouts, domain mapping of MAVS-MKK7 interaction, in vivo mouse data","pmids":["24651600"],"is_preprint":false},{"year":2014,"finding":"5Z-7-Oxozeaenol covalently binds to MAP2K7 at Cys218 (located at the end of the hinge region), not at the gatekeeper-2 cysteine, as revealed by crystal structure of the MAP2K7/5Z7O complex.","method":"X-ray crystallography","journal":"Bioorganic & medicinal chemistry letters","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure defining covalent binding site, single study","pmids":["25529738"],"is_preprint":false},{"year":2015,"finding":"NMR spectroscopy revealed the conformational behavior of the MKK7 regulatory domain: three docking sites (D1, D2, D3) show diverse intrinsic conformational propensities and different interaction kinetics with JNK1, though similar affinities. Crystal structure of JNK1 in complex with the second D-site of MKK7 revealed two different binding modes correlating with NMR exchange spectroscopy observations.","method":"NMR spectroscopy (atomic-resolution ensemble), X-ray crystallography","journal":"PNAS","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus NMR, two orthogonal structural methods providing mechanistic insight into docking","pmids":["25737554"],"is_preprint":false},{"year":2015,"finding":"MKK7 alternative splicing during T-cell activation (skipping exon 2, mediated by CELF2 RNA-binding protein) restores a JNK-docking site disrupted in the larger isoform, enhancing JNK pathway activity (c-Jun phosphorylation, TNF-α upregulation). This splicing event is itself JNK-dependent, creating a positive feedback loop.","method":"Alternative splicing analysis, CELF2 knockdown/overexpression, phospho-immunoblot, CLIP/RNA binding assay","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — mechanistic feedback loop established with genetic manipulation of CELF2 and splicing, multiple orthogonal readouts","pmids":["26443849"],"is_preprint":false},{"year":2015,"finding":"Knockdown of MKK7 or ASK1 blocked DSS-induced tight junction disruption and barrier dysfunction. A Ca2+/Ask1/MKK7/JNK2/c-Src signaling cascade mediates DSS-induced tight junction disruption; JNK2-dependent Src activation leads to tyrosine phosphorylation of junctional proteins.","method":"siRNA knockdown, intracellular Ca2+ measurement, in vitro kinase assay with recombinant JNK2 and c-Src, mouse colitis model","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with in vitro kinase assay, in vivo model, single lab","pmids":["25377781"],"is_preprint":false},{"year":2016,"finding":"DUSP22 acts as a scaffold protein for the ASK1–MKK7–JNK signal transduction pathway by selectively associating with ASK1, MKK7, and JNK1/2; it increases JNK phosphorylation independently of its phosphatase activity.","method":"Co-immunoprecipitation, overexpression of phosphatase-dead mutant, JNK kinase assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP and phosphatase-dead mutant, single lab; scaffold behavior supported by dose-response analysis","pmids":["27711255"],"is_preprint":false},{"year":2016,"finding":"KLF4 transcriptionally represses the MAP2K7 gene. In T-ALL cells with KLF4 inactivation, aberrant activation of MAP2K7 and downstream JNK and ATF2 occurs. JNK inhibitors reduce expansion of leukemia cells in patient-derived xenograft models.","method":"ChIP/gene expression, CRISPR/shRNA knockdown, xenograft model, phospho-immunoblot","journal":"Leukemia","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis by KLF4 loss-of-function with MAP2K7 activation readout, single lab","pmids":["27872496"],"is_preprint":false},{"year":2018,"finding":"SENP3-mediated deSUMOylation of MKK7 (SUMO2/3 removal) favors MKK7 binding to JNK and potentiates LPS-induced JNK phosphorylation and inflammatory cytokine production. SENP3 deficiency impairs MKK7 activity and JNK phosphorylation; ROS-dependent SENP3 accumulation after LPS drives MKK7 deSUMOylation.","method":"Conditional knockout mice, co-immunoprecipitation, in vivo and in vitro SUMO assay, phospho-immunoblot, septic shock model","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional knockout, co-IP, in vivo SUMOylation assay, multiple orthogonal methods","pmids":["29352108"],"is_preprint":false},{"year":2018,"finding":"Covalent virtual screening identified selective covalent MKK7 inhibitors. Crystal structure of lead compound bound to MKK7 confirmed the predicted binding mode. Inhibitors block JNK phosphorylation in cells, validated using MKK7 knockout cell lines, and block B-cell activation by LPS.","method":"Covalent virtual screening, X-ray crystallography, kinase selectivity panel (76 kinases), proteomics, knockout cell line validation","journal":"Cell chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure, selectivity panel, knockout validation, multiple orthogonal approaches","pmids":["30449673"],"is_preprint":false},{"year":2018,"finding":"Chemical cross-linking mass spectrometry showed that the GADD45β–MKK7 interaction largely occurs between GADD45β loop 2 (residues 103–117) and the MKK7 kinase enzymatic pocket. DTP3 disrupts this interaction by contacting MKK7 peptides 113–136 and 259–274.","method":"Chemical cross-linking mass spectrometry (CX-MS), enzymatic MS footprinting, fluorescence quenching with truncation mutants","journal":"International journal of biological macromolecules","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — mass spectrometry-based structural characterization with mutagenesis validation, single lab","pmids":["29572137"],"is_preprint":false},{"year":2020,"finding":"Crystal structures of MKK7 revealed the structural basis of catalytic domain plasticity and the role of the N-terminal regulatory helix in mediating kinase activation (active-state structure). An allosteric pocket in the N-terminal lobe was identified where ibrutinib binds. Type II irreversible inhibitor binding mode and multiple novel scaffolds were identified.","method":"X-ray crystallography (comprehensive set of structures), small-molecule screening, biochemical kinase assays","journal":"Cell chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — comprehensive crystal structure set including first MAP2K active state model, multiple compounds validated","pmids":["32783966"],"is_preprint":false},{"year":2020,"finding":"A MAP2K7 splice variant lacking exon 2 (MAP2K7Δexon2), induced by loss of MBNL1, activates JNK and promotes tumor dedifferentiation. This is the same exon-2-skipping isoform favored during T-cell activation; JNK inhibition reverses MAP2K7Δexon2-driven dedifferentiation.","method":"Alternative splicing analysis (TCGA/cancer genome), MBNL1 knockdown/overexpression, JNK inhibitor treatment, in vitro and in vivo tumor assays","journal":"PNAS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mechanistic splice-isoform validated in vitro and in vivo, single lab, corroborated by prior splicing work","pmids":["32601196"],"is_preprint":false},{"year":2022,"finding":"EEF1AKMT3 methyltransferase catalyzes monomethylation of MAP2K7 at K296, which decreases MAP2K7 phosphorylation, ubiquitination, and degradation of TP53. Loss of EEF1AKMT3 leads to MAP2K7 activation, increased tumor invasiveness and migration in gastric cancer.","method":"Mass spectrometry (substrate identification), gain/loss-of-function studies, phospho-antibody array, RNA-seq","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — mass spectrometry identification of methylation site with functional gain/loss studies, single lab","pmids":["35753528"],"is_preprint":false},{"year":2015,"finding":"MKK7 neddylation (by RanBP2ΔFG E3 ligase) reduces its basal kinase activity. RanBP2 knockdown increases MKK7 kinase activity; ectopic RanBP2ΔFG reduces it. In vitro neddylation of purified MKK7 by RanBP2ΔFG directly reduces basal kinase activity.","method":"Co-immunoprecipitation, in vitro neddylation assay, kinase activity assay, siRNA knockdown","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 1-2 / Moderate — in vitro neddylation reconstitution with direct kinase activity measurement, single lab","pmids":["26364603"],"is_preprint":false},{"year":2010,"finding":"RASSF7 interacts with N-Ras and MKK7 to negatively regulate JNK signaling. RASSF7 promotes the phosphorylated state of MKK7 but inhibits its ability to activate JNK. RASSF7 requires its RA domain for interaction with GTP-bound N-Ras and the anti-apoptotic effect. RASSF7 undergoes proteasomal degradation under prolonged stress, allowing MKK7/JNK death signaling to proceed.","method":"Co-immunoprecipitation, RNAi knockdown, kinase assay, domain-deletion analysis, ubiquitin-proteasome pathway assay","journal":"Cell death and differentiation","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, RNAi, kinase assay, domain analysis; single lab","pmids":["21278800"],"is_preprint":false},{"year":2017,"finding":"Neuron-specific MKK7 knockout mice display age-dependent motor dysfunction, axonal degeneration in the spinal cord, and accumulation of axonal transport proteins JIP1 and APP in brain and spinal cord, establishing that MKK7-JNK signaling is required for neuronal maintenance and axonal transport in adult neurons.","method":"Conditional knockout (neuron-specific), behavioral analysis, histology, immunoblot for axonal transport proteins","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional knockout with defined molecular markers, single lab","pmids":["28779160"],"is_preprint":false}],"current_model":"MAP2K7 (MKK7) is a dual-specificity MAPK kinase that preferentially phosphorylates the Thr183 residue (within the Thr-Pro-Tyr motif) of JNK isoforms to promote their activation; it acts synergistically and non-redundantly with MKK4 (which preferentially phosphorylates Tyr185) for full JNK activation. MKK7 is the principal JNK activator downstream of proinflammatory cytokines (TNFα, IL-1), Fas, MAVS/antiviral signaling, and several MAP3Ks (DLK, MLK2, MLK3, MEKK2/3). Its N-terminal regulatory domain contains three cooperative JNK-docking D-sites and is subject to alternative splicing (exon 2 exclusion enhances JNK docking). MKK7 activity is regulated by multiple PTMs—inhibitory SUMOylation (reversed by SENP3) and neddylation (by RanBP2), and activating monomethylation at K296 by EEF1AKMT3—and by direct binding partners including GADD45β (blocks its ATP site), c-FLIPL (inhibits upstream kinase recruitment), RASSF7/N-Ras (inhibits downstream JNK activation), Filamin A (scaffolds MKK7 and MKK4), DUSP22 (scaffold for ASK1–MKK7–JNK), and TIPRL (recruits PP2Ac to dephosphorylate/inactivate MKK7). Structurally, MKK7 has been characterized by crystal structures revealing its catalytic domain plasticity, an N-terminal regulatory helix required for activation, a covalent inhibitor binding site at Cys218, and an allosteric N-lobe pocket. Physiologically, MKK7 is essential for JNK-dependent cell cycle progression (G2/M via CDC2/c-Jun), tumor suppression (coupling oncogenic stress to p53 stability), axon elongation and neuronal maintenance, cardiomyocyte stress adaptation, and negative regulation of hematopoietic cell proliferation."},"narrative":{"mechanistic_narrative":"MAP2K7 (MKK7) is a JNK-specific dual-specificity MAP kinase kinase that constitutes the principal stress- and cytokine-activated arm of the SAPK/JNK signaling module [PMID:9384583, PMID:9372971]. It directly phosphorylates and activates JNK/SAPK without engaging p38 or ERK, and it does so non-redundantly with MKK4: MKK7 selectively phosphorylates the threonine (Thr183) of the JNK activation loop while MKK4 targets the tyrosine (Tyr185), so the two kinases act synergistically to achieve full dual phosphorylation and maximal JNK activity [PMID:11062067, PMID:10715136, PMID:11418587]. Genetic disruption established that MKK7 alone is required for JNK activation by proinflammatory cytokines, whereas combined loss of MKK4 and MKK7 is needed to block stress-induced activation [PMID:11390361]. MKK7 sits downstream of diverse stimuli—TNFα, IL-1, Fas, Rac1, and antiviral MAVS signaling—and of multiple MAP3Ks including DLK, MLK2/MST, MLK3, and MEKK2/3 that preferentially feed MKK7 over MKK4 [PMID:9362518, PMID:9414114, PMID:10187804, PMID:10347227, PMID:9516438, PMID:24651600]. Substrate engagement is governed by an N-terminal regulatory domain bearing three cooperative JNK-docking D-sites, and alternative splicing of exon 2 modulates this docking to tune pathway output [PMID:16533805, PMID:25737554, PMID:26443849]. MKK7 activity is constrained by an array of direct regulators: GADD45β binds and blocks its catalytic/ATP pocket downstream of NF-κB [PMID:14743220, PMID:17485467], c-FLIPL blocks upstream MAP3K recruitment [PMID:17110930], TIPRL recruits PP2A to dephosphorylate it [PMID:22841785], and SUMOylation (reversed by SENP3) and RanBP2-mediated neddylation impose additional negative control [PMID:29352108, PMID:26364603], while Filamin A and DUSP22 scaffold MKK7 with its upstream and downstream partners [PMID:20156194, PMID:27711255]. Through JNK–c-Jun, MKK7 couples oncogenic and genotoxic stress to p53 stability and CDC2-dependent G2/M cell cycle control, acting as a tumor suppressor and a negative regulator of hematopoietic proliferation via the p16INK4a axis [PMID:15039780, PMID:21317887, PMID:11560992]. It is additionally essential for axon elongation, adult neuronal maintenance and axonal transport, and cardiomyocyte stress adaptation [PMID:22090513, PMID:28779160, PMID:21284947]. Crystallographic and NMR studies have defined its catalytic-domain plasticity, an activating N-terminal regulatory helix, a covalent inhibitor site at Cys218, and an allosteric N-lobe pocket [PMID:25529738, PMID:32783966, PMID:25737554].","teleology":[{"year":1997,"claim":"Establishing that a dedicated JNK-specific MAPKK existed answered whether JNK activation required a kinase distinct from the p38/JNK dual activator MKK4.","evidence":"Molecular cloning of MKK7/JNKK2 with in vitro kinase assays and Drosophila Hemipterous complementation across human and murine systems","pmids":["9384583","9312105","9372971"],"confidence":"High","gaps":["Site-specificity of MKK7 versus MKK4 on the JNK activation loop not yet resolved","In vivo non-redundancy not yet demonstrated genetically"]},{"year":1997,"claim":"Mapping MKK7 to specific receptor inputs showed it is the JNK activator selectively engaged by death-receptor and cytokine signaling rather than a generic stress kinase.","evidence":"Immune-complex kinase assays with isoform-specific antisera in Fas-stimulated cells and IL-1-stimulated rabbit liver","pmids":["9362518","9414114"],"confidence":"Medium","gaps":["Direct upstream MAP3K linking receptors to MKK7 not identified","Single-lab observations per stimulus"]},{"year":1999,"claim":"Identifying alternatively spliced isoforms and a JNK-binding N-terminal extension explained how MKK7 docks its substrate and how basal versus inducible activity is tuned.","evidence":"Molecular cloning of six isoforms with direct binding and kinase assays plus subcellular localization","pmids":["9891090"],"confidence":"High","gaps":["Physiological triggers selecting specific isoforms not defined","Structural basis of docking not yet resolved"]},{"year":1999,"claim":"Defining the MAP3K tier showed that DLK, MLK2, and MEKK2/3 preferentially activate MKK7 over MKK4, establishing pathway selectivity at the kinase-cascade level.","evidence":"In vitro reconstitution kinase assays with recombinant proteins, co-IP, and column fractionation","pmids":["10187804","10347227","9516438"],"confidence":"High","gaps":["Determinants of MAP3K substrate preference partly defined only for MLK2","Quantitative contribution of each MAP3K in vivo unknown"]},{"year":2000,"claim":"Demonstrating that MKK7 phosphorylates Thr183 while MKK4 phosphorylates Tyr185 resolved the mechanistic basis of their synergistic, non-redundant cooperation.","evidence":"In vitro kinase assays with phosphoamino-acid analysis and mass spectrometry across JNK isoforms","pmids":["11062067","10715136"],"confidence":"High","gaps":["Whether sequential ordering of the two phosphorylations is obligatory not fully settled","Isoform-specific kinetics in cells not measured"]},{"year":2001,"claim":"Genetic knockout established the in vivo division of labor: MKK7 is essential for cytokine-induced JNK activation, while stress activation requires both MKK7 and MKK4.","evidence":"Targeted Mkk7 and Mkk4/Mkk7 gene disruption in mice with phospho-specific JNK readouts","pmids":["11390361","11418587"],"confidence":"High","gaps":["Cell-type-specific differences in input dependence not fully mapped","Isoform-specific contributions not separable by knockout"]},{"year":2001,"claim":"Conditional deletion revealed MKK7 as a negative regulator of hematopoietic proliferation acting through the p16INK4a cell-cycle brake.","evidence":"Conditional gene targeting in thymocytes, B cells, and mast cells with p16INK4a reexpression rescue","pmids":["11560992"],"confidence":"High","gaps":["Direct molecular link between JNK signaling and p16INK4a induction not defined","Whether effect generalizes beyond hematopoietic lineages unknown"]},{"year":2004,"claim":"Linking MKK7-JNK-c-Jun to CDC2 placed the pathway directly in control of G2/M progression and senescence, framing its cell-cycle function.","evidence":"Gene knockout in MEFs with cell cycle analysis and c-Jun mutant epistasis","pmids":["15039780"],"confidence":"High","gaps":["Mechanism by which c-Jun regulates CDC2 not detailed","Connection to p16/p53 arms not integrated"]},{"year":2004,"claim":"Identifying GADD45β as a direct catalytic inhibitor explained how NF-κB suppresses JNK and protects against TNFα cytotoxicity.","evidence":"Unbiased screen, direct binding, kinase inhibition, and disrupting-peptide rescue; later refined by structural mapping of the ATP-pocket-engaging interface","pmids":["14743220","17485467","18343408","29572137"],"confidence":"High","gaps":["Stoichiometry and full structure of the complex defined only by prediction/NMR, not crystallography","Regulation of GADD45β abundance in vivo incompletely defined"]},{"year":2006,"claim":"Systematic mapping of three cooperative N-terminal D-sites established the structural logic of MKK7–JNK substrate docking.","evidence":"Site-directed mutagenesis of all three docking sites with quantitative binding and kinase assays, later extended by NMR and crystallography","pmids":["16533805","25737554"],"confidence":"High","gaps":["Functional consequence of distinct docking-site binding modes not fully resolved","Regulation of docking by phosphorylation/PTMs not addressed"]},{"year":2010,"claim":"Discovery of scaffolds and additional binding-partner regulators (Filamin A, RASSF7/N-Ras) showed how MKK7 is spatially organized and its output set by competing inputs.","evidence":"Co-IP, loss-of-function cell lines, domain mapping, and colocalization","pmids":["20156194","21278800"],"confidence":"Medium","gaps":["RASSF7 mechanism (phospho-MKK7 yet inhibited JNK output) not mechanistically explained","Scaffold contributions in vivo not tested genetically"]},{"year":2011,"claim":"In vivo tumor models established MKK7 as a tumor suppressor coupling oncogenic stress to p53 stability through JNK1/2.","evidence":"Tissue-specific conditional knockout in KRas- and NeuT-driven tumor models with JNK1/JNK2 epistasis and p53 stability assays","pmids":["21317887"],"confidence":"High","gaps":["Molecular step connecting JNK to p53 stabilization not defined","Relevance to human tumors with MAP2K7 alterations not established here"]},{"year":2011,"claim":"Conditional deletion in neurons and cardiomyocytes defined tissue-specific physiological roles in axon elongation, neuronal maintenance, and cardiac stress adaptation.","evidence":"Nestin-Cre and cardiomyocyte-specific knockouts with histology, axon assays, pressure-overload surgery, and JNK substrate immunoblots","pmids":["22090513","21284947","28779160"],"confidence":"High","gaps":["Which JNK substrates dominate each phenotype not fully dissected","Cardiac and neuronal effects characterized in single labs"]},{"year":2012,"claim":"Local mRNA translation at the growth cone and a DLK–MKK7–JNK1 module showed MKK7 can act on cytoskeletal substrates (MAP1B) spatially uncoupled from nuclear transcription.","evidence":"Genome-wide neurite mRNA screen, mRNA localization imaging, and kinase-module manipulation","pmids":["23226105"],"confidence":"High","gaps":["Regulation of localized translation by upstream signals not defined","Generality beyond neurons unknown"]},{"year":2014,"claim":"Identifying MAVS-mediated mitochondrial recruitment and selective JNK2 phosphorylation defined an antiviral apoptotic function for MKK7.","evidence":"Multiple genetic knockouts, MAVS domain mapping, mitochondrial fractionation, and viral challenge in mice","pmids":["24651600"],"confidence":"High","gaps":["Basis for JNK2 versus JNK1 selectivity not mechanistically resolved","Interplay with other antiviral pathways not addressed"]},{"year":2014,"claim":"Druggability was established by targeting the GADD45β/MKK7 interface and by covalent occupancy of Cys218, opening therapeutic and chemical-biology avenues.","evidence":"DTP3 complex-disruption assays with xenografts, plus crystal structures of covalent inhibitor complexes and selective inhibitor development validated in knockout cells","pmids":["25314077","25529738","30449673","29572137"],"confidence":"High","gaps":["Clinical translation of these agents not addressed in the corpus","Off-target/selectivity profiles defined only for specific scaffolds"]},{"year":2015,"claim":"A layered PTM code (SUMOylation/SENP3, RanBP2 neddylation, EEF1AKMT3 methylation) and CELF2/MBNL1-controlled exon-2 splicing revealed how MKK7 activity is set post-translationally and post-transcriptionally.","evidence":"Conditional knockouts, in vitro neddylation/SUMO assays, mass-spectrometric methylation-site identification, and splicing manipulation","pmids":["26443849","29352108","26364603","35753528","32601196"],"confidence":"High","gaps":["Hierarchy and crosstalk among the different PTMs not integrated","In vivo importance of each modification context-limited"]},{"year":2020,"claim":"Comprehensive crystal structures defined catalytic-domain plasticity, an activating N-terminal regulatory helix, and an allosteric N-lobe pocket, providing a structural framework for MKK7 activation and inhibition.","evidence":"Comprehensive X-ray crystallography including an active-state model plus small-molecule screening and kinase assays","pmids":["32783966"],"confidence":"High","gaps":["Structure of full-length MKK7 with regulatory domain and docked JNK not solved","Conformational dynamics during the activation cycle not fully captured"]},{"year":null,"claim":"How the multiple negative regulators, PTMs, scaffolds, and isoforms are integrated to set MKK7 output in a given cell type and stimulus, and how this is therapeutically exploitable beyond GADD45β disruption, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified quantitative model of MKK7 regulation across inputs","Context-specific dominance of each regulator unknown","In vivo consequences of combined PTM perturbation untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,13,14]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,13,18]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[23,29,43]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[7,21]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[7,11]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[42]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[33,38]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,13,15]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[4,15,28,49]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[24,34]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[23,42,56]}],"complexes":["MKK4–MKK7–JNK complex","ASK1–MKK7–JNK (DUSP22-scaffolded)","GADD45β–MKK7 inhibitory complex"],"partners":["MAPK8","MAP2K4","GADD45B","MAP3K12","FLNA","TIPRL","DUSP22","RASSF7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O14733","full_name":"Dual specificity mitogen-activated protein kinase kinase 7","aliases":["JNK-activating kinase 2","MAPK/ERK kinase 7","MEK 7","Stress-activated protein kinase kinase 4","SAPK kinase 4","SAPKK-4","SAPKK4","c-Jun N-terminal kinase kinase 2","JNK kinase 2","JNKK 2"],"length_aa":419,"mass_kda":47.5,"function":"Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. Essential component of the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. With MAP2K4/MKK4, is the one of the only known kinase to directly activate the stress-activated protein kinase/c-Jun N-terminal kinases MAPK8/JNK1, MAPK9/JNK2 and MAPK10/JNK3. MAP2K4/MKK4 and MAP2K7/MKK7 both activate the JNKs by phosphorylation, but they differ in their preference for the phosphorylation site in the Thr-Pro-Tyr motif. MAP2K4/MKK4 shows preference for phosphorylation of the Tyr residue and MAP2K7/MKK7 for the Thr residue. The monophosphorylation of JNKs on the Thr residue is sufficient to increase JNK activity indicating that MAP2K7/MKK7 is important to trigger JNK activity, while the additional phosphorylation of the Tyr residue by MAP2K4/MKK4 ensures optimal JNK activation. Has a specific role in JNK signal transduction pathway activated by pro-inflammatory cytokines. The MKK/JNK signaling pathway is also involved in mitochondrial death signaling pathway, including the release cytochrome c, leading to apoptosis. Part of a non-canonical MAPK signaling pathway, composed of the upstream MAP3K12 kinase and downstream MAP kinases MAPK1/ERK2 and MAPK3/ERK1, that enhances the AP-1-mediated transcription of APP in response to APOE (PubMed:28111074)","subcellular_location":"Nucleus; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/O14733/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MAP2K7","classification":"Not Classified","n_dependent_lines":54,"n_total_lines":1208,"dependency_fraction":0.04470198675496689},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000076984","cell_line_id":"CID001194","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"ANAPC11","stoichiometry":4.0},{"gene":"PDLIM7","stoichiometry":4.0},{"gene":"C14ORF1","stoichiometry":0.2},{"gene":"C19ORF24","stoichiometry":0.2},{"gene":"MOCS2","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID001194","total_profiled":1310},"omim":[{"mim_id":"618642","title":"SH3 DOMAIN-CONTAINING RING FINGER PROTEIN 1; SH3RF1","url":"https://www.omim.org/entry/618642"},{"mim_id":"611437","title":"DUAL-SPECIFICITY PHOSPHATASE 19; DUSP19","url":"https://www.omim.org/entry/611437"},{"mim_id":"610105","title":"MICRO RNA 125B2; MIR125B2","url":"https://www.omim.org/entry/610105"},{"mim_id":"609487","title":"MITOGEN-ACTIVATED PROTEIN KINASE KINASE KINASE 2; MAP3K2","url":"https://www.omim.org/entry/609487"},{"mim_id":"609479","title":"MITOGEN-ACTIVATED PROTEIN KINASE KINASE KINASE 20; MAP3K20","url":"https://www.omim.org/entry/609479"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Plasma membrane","reliability":"Uncertain"},{"location":"Cytosol","reliability":"Uncertain"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MAP2K7"},"hgnc":{"alias_symbol":["MKK7","Jnkk2","SAPKK4"],"prev_symbol":["PRKMK7"]},"alphafold":{"accession":"O14733","domains":[{"cath_id":"3.30.200.20","chopping":"96-195","consensus_level":"high","plddt":84.0191,"start":96,"end":195},{"cath_id":"1.10.510.10","chopping":"200-399","consensus_level":"high","plddt":88.8943,"start":200,"end":399}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O14733","model_url":"https://alphafold.ebi.ac.uk/files/AF-O14733-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O14733-F1-predicted_aligned_error_v6.png","plddt_mean":77.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MAP2K7","jax_strain_url":"https://www.jax.org/strain/search?query=MAP2K7"},"sequence":{"accession":"O14733","fasta_url":"https://rest.uniprot.org/uniprotkb/O14733.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O14733/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O14733"}},"corpus_meta":[{"pmid":"11390361","id":"PMC_11390361","title":"MKK7 is an essential component of the JNK signal transduction pathway activated by proinflammatory cytokines.","date":"2001","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/11390361","citation_count":310,"is_preprint":false},{"pmid":"14743220","id":"PMC_14743220","title":"Gadd45 beta mediates the NF-kappa B suppression of JNK signalling by targeting MKK7/JNKK2.","date":"2004","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/14743220","citation_count":292,"is_preprint":false},{"pmid":"9384583","id":"PMC_9384583","title":"A novel SAPK/JNK kinase, MKK7, stimulated by TNFalpha and cellular stresses.","date":"1997","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/9384583","citation_count":232,"is_preprint":false},{"pmid":"9575191","id":"PMC_9575191","title":"T lymphocyte activation signals for interleukin-2 production involve activation of MKK6-p38 and MKK7-SAPK/JNK signaling pathways sensitive to cyclosporin A.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9575191","citation_count":178,"is_preprint":false},{"pmid":"11062067","id":"PMC_11062067","title":"Synergistic activation of stress-activated protein kinase 1/c-Jun N-terminal kinase (SAPK1/JNK) isoforms by mitogen-activated protein kinase kinase 4 (MKK4) and MKK7.","date":"2000","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/11062067","citation_count":164,"is_preprint":false},{"pmid":"9312105","id":"PMC_9312105","title":"MKK7 is a stress-activated mitogen-activated protein kinase kinase functionally related to hemipterous.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9312105","citation_count":155,"is_preprint":false},{"pmid":"9891090","id":"PMC_9891090","title":"The MKK7 gene encodes a group of c-Jun NH2-terminal kinase kinases.","date":"1999","source":"Molecular and cellular 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communications","url":"https://pubmed.ncbi.nlm.nih.gov/18515079","citation_count":21,"is_preprint":false},{"pmid":"29572137","id":"PMC_29572137","title":"Probing the interaction interface of the GADD45β/MKK7 and MKK7/DTP3 complexes by chemical cross-linking mass spectrometry.","date":"2018","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/29572137","citation_count":20,"is_preprint":false},{"pmid":"17187786","id":"PMC_17187786","title":"Differential requirement of MKK4 and MKK7 in JNK activation by distinct scaffold proteins.","date":"2006","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/17187786","citation_count":20,"is_preprint":false},{"pmid":"35234473","id":"PMC_35234473","title":"Circular RNA ACTA1 Acts as a Sponge for miR-199a-5p and miR-433 to Regulate Bovine Myoblast Development through the MAP3K11/MAP2K7/JNK Pathway.","date":"2022","source":"Journal of agricultural and food chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/35234473","citation_count":19,"is_preprint":false},{"pmid":"28779160","id":"PMC_28779160","title":"Age-dependent motor dysfunction due to neuron-specific disruption of stress-activated protein kinase MKK7.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28779160","citation_count":19,"is_preprint":false},{"pmid":"30768270","id":"PMC_30768270","title":"Targeting the MKK7-JNK (Mitogen-Activated Protein Kinase Kinase 7-c-Jun N-Terminal Kinase) Pathway with Covalent Inhibitors.","date":"2019","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30768270","citation_count":19,"is_preprint":false},{"pmid":"27526795","id":"PMC_27526795","title":"Heme Oxygenase-1 Inhibits Neuronal Apoptosis in Spinal Cord Injury through Down-Regulation of Cdc42-MLK3-MKK7-JNK3 Axis.","date":"2016","source":"Journal of neurotrauma","url":"https://pubmed.ncbi.nlm.nih.gov/27526795","citation_count":19,"is_preprint":false},{"pmid":"10873670","id":"PMC_10873670","title":"Stress-activated protein kinase-dependent induction of c-fos by Cd(2+) is mediated by MKK7.","date":"2000","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/10873670","citation_count":19,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":53715,"output_tokens":13894,"usd":0.184778,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":25722,"output_tokens":5931,"usd":0.138442,"stage2_stop_reason":"end_turn"},"total_usd":0.32322,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1997,\n      \"finding\": \"MKK7 (JNKK2) was cloned and identified as a novel MAPKK that is specific for the SAPK/JNK subgroup; unlike SEK1/MKK4, it does not activate p38. MKK7 directly phosphorylates and activates JNK/SAPK. It is activated by TNFα and environmental stresses, and immunochemical studies identified MKK7 as a major JNK-activating kinase in osmotically shocked cells. Overexpression of MKK7 enhanced AP-1-dependent transcription.\",\n      \"method\": \"Molecular cloning, immunoprecipitation, in vitro kinase assay, co-expression studies, reporter assay\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — original cloning paper with in vitro kinase assay, substrate specificity demonstrated, replicated across subsequent papers\",\n      \"pmids\": [\"9384583\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"MKK7 was identified as a murine homolog of Drosophila Hemipterous; it functionally rescues hep mutant flies. In fibroblasts, MKK7 is activated by stress and by GTPase Rac1. MKK7 directly phosphorylates and activates JNK/SAPK.\",\n      \"method\": \"Molecular cloning, Drosophila complementation rescue, in vitro kinase assay, transfection in fibroblasts\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay plus in vivo complementation, replicated by multiple labs\",\n      \"pmids\": [\"9312105\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Human JNKK2 (MKK7) was cloned and shown to be a highly specific JNK kinase that does not activate p38 MAPK. Unlike JNKK1/MKK4, it is JNK-specific.\",\n      \"method\": \"Database search, cDNA cloning, co-expression kinase assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — direct cloning and biochemical characterization, replicated by multiple labs\",\n      \"pmids\": [\"9372971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"MKK7 (but not SEK1/MKK4) is activated by Fas signaling as the upstream activator of JNK/SAPK; MKK6 is the major p38 activator in this pathway. JNK/SAPK activation in Fas signaling does not require CPP32-like proteases.\",\n      \"method\": \"Immune complex kinase assay, peptide inhibitor pharmacology, immunoprecipitation\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — kinase assay with multiple inhibitor controls, single lab\",\n      \"pmids\": [\"9362518\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"MKK7 is the activator of JNK/SAPK activated by IL-1 in rabbit liver; it is not activated by MKK4 antibodies. JNK/SAPK is the only MAPK activated by IL-1 in liver.\",\n      \"method\": \"S-Sepharose purification, immunoprecipitation with anti-MKK7 and anti-MKK4 antisera, kinase assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — immunoprecipitation and kinase assay, single lab, replicated conceptually by others\",\n      \"pmids\": [\"9414114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Human MKK7 (47 kDa) specifically phosphorylates and activates JNK1 but fails to activate p38 MAPK in co-expression studies. MKK7 is activated in hematopoietic cells by IL-3, CD40 ligation, B-cell antigen receptor, FcR, heat, UV, anisomycin, hyperosmolarity, TNFα, and by constitutively active Ras, Rac, and Cdc42.\",\n      \"method\": \"Molecular cloning, co-expression, immunoprecipitation kinase assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — direct biochemical characterization with substrate specificity, replicated across labs\",\n      \"pmids\": [\"9535930\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"MKK7 plays a major role in SAPK/JNK activation in T lymphocytes responding to TCR/co-stimulation. Dominant-negative MKK7 abrogated transcriptional activation of the distal NFAT response element in the IL-2 promoter. Both the MKK6-p38 and MKK7-JNK pathways are activated in a cyclosporin A-sensitive manner and contribute to IL-2 gene expression.\",\n      \"method\": \"Dominant-negative transfection, reporter assay, kinase assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dominant-negative and reporter assay, single lab, conceptually replicated\",\n      \"pmids\": [\"9575191\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"The MKK7 gene encodes six isoforms (α1/2, β1/2, γ1/2) by alternative splicing with three different N-termini and two C-termini. The N-terminal extension (absent in MKK7α) binds directly to the MKK7 substrate JNK. MKK7α isoforms show lower basal activity but higher inducible fold-activation than β and γ isoforms. MKK7 is detected in both cytoplasmic and nuclear compartments, but nuclear localization is not required for JNK activation in vivo.\",\n      \"method\": \"Molecular cloning, yeast two-hybrid/direct binding, kinase assay, immunofluorescence\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — molecular cloning with direct binding and kinase assays, multiple orthogonal methods\",\n      \"pmids\": [\"9891090\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"DLK (dual leucine zipper-bearing kinase) associates with, phosphorylates, and activates MKK7 in vitro and in vivo, but unlike MLK3 does not phosphorylate or activate MKK4. DLK and MKK7 co-localize in neurons and occupy similar subcellular compartments, distinct from MKK4.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, immunocytochemistry, subcellular fractionation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — in vitro kinase assay, reciprocal co-IP, and subcellular fractionation with functional consequence\",\n      \"pmids\": [\"10187804\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"MEKK3 (and MEKK2) directly activates MKK7 and MKK6 in vitro; immunoprecipitates of MEKK3 phosphorylate recombinant MKK7 in vitro. Coexpression of MKK7 with MEKK3 in COS-7 cells enhanced MKK7 autophosphorylation and its ability to activate JNK1.\",\n      \"method\": \"Co-expression, in vitro kinase assay with recombinant proteins, immunoprecipitation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with recombinant proteins plus cellular co-expression, single lab\",\n      \"pmids\": [\"10347227\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"MST/MLK2 activates recombinant MKK7 more effectively than recombinant SEK1/MKK4 in vitro. The majority of MLK2-dependent JNK-activating activity co-fractionates with MKK7, not MKK4. The MLK2 kinase domain determines this substrate specificity.\",\n      \"method\": \"In vitro reconstitution kinase assay, column fractionation, Western blot\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay with deletion analysis, single lab\",\n      \"pmids\": [\"9516438\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"A JNKK2-JNK1 fusion protein acts as a constitutively active Jun kinase; JNK1 is phosphorylated by JNKK2 at both Thr183 and Tyr185 in the fusion. The fusion protein is specific for the JNK pathway (does not activate p38 or ERK2) and is sufficient to stimulate c-Jun transcriptional activity. The fusion protein localizes predominantly to the nucleus.\",\n      \"method\": \"Fusion protein construction, immunoblotting with phospho-specific antibodies, in vitro kinase assay, reporter assay, immunofluorescence\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal methods including phospho-specific immunoblot, kinase assay, and transcriptional reporter\",\n      \"pmids\": [\"10506143\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"G protein βγ subunit activates MKK7 by ~2-fold in a Rac-dependent manner; this is distinct from Gbγ-induced MKK4 activation which depends on Rho and Cdc42. MKK7 activation by Gbγ is not blocked by tyrosine kinase inhibitors PP2/PP1, unlike MKK4 activation.\",\n      \"method\": \"Transfection with kinase-deficient mutants, dominant-negative GTPases, kinase activity assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dominant-negative epistasis and kinase assay, single lab\",\n      \"pmids\": [\"9890951\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"MKK4 shows a striking preference for phosphorylating Tyr185 of SAPK1/JNK isoforms, while MKK7 shows a striking preference for Thr183. Together, MKK4 and MKK7 produce a synergistic increase in JNK activity in vitro. MKK7β is several hundred-fold more efficient than MKK7α' in activating JNK isoforms.\",\n      \"method\": \"In vitro kinase assay, phosphoamino acid analysis, mass spectrometry\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — rigorous in vitro reconstitution with site-specific phosphorylation analysis across multiple JNK isoforms\",\n      \"pmids\": [\"11062067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"MKK7 monophosphorylates JNK3α1 at Thr183 (Thr residue) in vitro; both MKK4 and MKK7 are required for bisphosphorylation and maximal enzyme activity of JNK3α1. MKK4 alone shows no phosphorylation of JNK3α1 by mass spectrometry.\",\n      \"method\": \"In vitro kinase assay, mass spectral phosphorylation analysis, kinetic measurements\",\n      \"journal\": \"Biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — mass spectrometry-verified phosphorylation sites with kinetic characterization\",\n      \"pmids\": [\"10715136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Genetic disruption of Mkk7 alone was sufficient to prevent JNK activation by proinflammatory cytokines, whereas simultaneous disruption of Mkk4 and Mkk7 was required to block JNK activation by environmental stress. MKK7 preferentially phosphorylates JNK on Thr and MKK4 on Tyr.\",\n      \"method\": \"Targeted gene disruption (knockout mice), JNK kinase assay, phospho-specific analysis\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knockout with biochemical readout, demonstrates non-redundant in vivo roles, replicated by multiple labs\",\n      \"pmids\": [\"11390361\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"MKK7 is an essential and specific regulator of SAPK/JNK activation in hematopoietic cells. Loss of MKK7 in thymocytes, mature B cells, and mast cells causes hyperproliferation; SAPK/JNK activation was completely abolished in mkk7−/− mast cells despite normal MKK4 phosphorylation. MKK7 negatively regulates proliferation through a pathway involving p16INK4a; reexpression of p16INK4a abrogates the hyperproliferative response.\",\n      \"method\": \"Conditional gene targeting, kinase assay, flow cytometry, Western blotting, reexpression rescue\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout with mechanistic rescue experiment (p16 reexpression), replicated across multiple cell types\",\n      \"pmids\": [\"11560992\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"PKCδ mediates ionizing radiation-induced JNK activation through MKK7 (not MKK4) in human thyroid cells. IR activates MKK7 but not MKK4; this was blocked by the PKCδ inhibitor rottlerin and by kinase-deficient MKK7, defining a PKCδ→MKK7→JNK→AP-1 cascade.\",\n      \"method\": \"Dominant-negative adenoviral expression, immune-complex kinase assay, pharmacological inhibitor\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — kinase assay with dominant-negative and pharmacological inhibitor, single lab\",\n      \"pmids\": [\"11314034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"SEK1 (MKK4) and MKK7 show synergistic SAPK/JNK activation in embryonic stem cells. MKK7 is responsible for Thr phosphorylation of JNK; without MKK7, Thr phosphorylation is lost but Tyr phosphorylation remains. MKK7α1 requires prior Tyr phosphorylation by SEK1 before it can phosphorylate JNK Thr; MKK7γ1 phosphorylates Thr independently; MKK7γ2 can phosphorylate both Thr and Tyr.\",\n      \"method\": \"Knockout ES cells, dual phosphorylation analysis, co-transfection with kinase-dead mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knockout combined with phospho-specific analyses and multiple isoform comparisons\",\n      \"pmids\": [\"11418587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"SKRP1, a novel dual-specificity phosphatase, interacts with MKK7 (co-precipitation in vitro and in vivo) and inactivates the JNK pathway by dephosphorylating JNK. SKRP1 does not bind directly to JNK but gains access to it through MKK7. SKRP1 does not interfere with MKK7-JNK co-precipitation.\",\n      \"method\": \"Co-immunoprecipitation, in vitro binding assay, in vitro phosphatase assay, overexpression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — co-IP in vitro and in vivo, phosphatase assay, single lab\",\n      \"pmids\": [\"11959861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"ZAK MAP3K utilizes MKK7 (not MKK4) to activate JNK/SAPK; dominant-negative MKK7 (but not MKK4) attenuated ZAK-induced JNK activation. ZAK activity disrupts actin stress fibers and causes G2/M cell cycle arrest.\",\n      \"method\": \"Co-expression, dominant-negative kinase mutants, flow cytometry, Western blot\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — dominant-negative overexpression only, single lab, single method\",\n      \"pmids\": [\"12220515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"In fibroblast-like synoviocytes, JNK, MKK4, and MKK7 form a stable complex detectable by co-immunoprecipitation, and MKK4 co-precipitates with MKK7. The complex localizes in the cytoplasm by confocal microscopy; JNK migrates to the nucleus after IL-1 stimulation. The complex is functionally active and phosphorylates c-Jun after IL-1.\",\n      \"method\": \"Co-immunoprecipitation, confocal microscopy, in vitro kinase assay\",\n      \"journal\": \"Arthritis and rheumatism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP and confocal colocalization with functional kinase readout, single lab\",\n      \"pmids\": [\"13130464\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"In mkk7−/− embryonic stem cells, Thr phosphorylation of JNK is lost while Tyr phosphorylation remains. In 293T cells, SEK1-induced Tyr phosphorylation of JNK1 is followed by additional Thr phosphorylation by MKK7. SEK1 (but not MKK7) binds to JNK1 in 293T cells, indicating sequential rather than independent phosphorylation.\",\n      \"method\": \"Knockout ES cells, phospho-specific immunoblot, co-immunoprecipitation, Thr-Pro-Phe JNK1 mutant\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — genetic knockout, co-IP, and mutant JNK analysis across two cell systems\",\n      \"pmids\": [\"12624093\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"GADD45β mediates NF-κB suppression of JNK signaling by directly binding MKK7 and blocking its catalytic activity. Gadd45β binds MKK7 directly; peptides disrupting the GADD45β/MKK7 interaction hinder the ability of both GADD45β and NF-κB to suppress TNFα-induced cytotoxicity.\",\n      \"method\": \"Unbiased screen, direct binding assay, kinase activity assay, peptide disruption, cell viability assay\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — direct binding demonstrated, kinase inhibition assayed, functional rescue with disrupting peptides\",\n      \"pmids\": [\"14743220\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"MKK7 couples stress signaling to G2/M cell cycle progression and cellular senescence. Genetic inactivation of MKK7 in MEFs leads to premature senescence and G2/M cell cycle arrest. The CDC2 kinase is a downstream target of the MKK7-JNK-c-Jun pathway; loss of c-Jun or JNK phosphorylation sites on c-Jun produces the same G2/M block.\",\n      \"method\": \"Gene knockout (MEFs), cell cycle analysis, epistasis with c-Jun mutants, CDC2 target identification\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout with defined cell cycle phenotype and pathway placement through c-Jun mutant epistasis\",\n      \"pmids\": [\"15039780\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"MLK3 activates MKK7 in the MLK3→MKK7→JNK cascade in hippocampal CA1 neurons after cerebral ischemia; K252a (MLK3 inhibitor) blocked MKK7 and JNK3 activation and was neuroprotective.\",\n      \"method\": \"In vivo ischemia model, immunoprecipitation kinase assay, pharmacological inhibition\",\n      \"journal\": \"Neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo kinase assay with pharmacological inhibitor, single lab\",\n      \"pmids\": [\"15680699\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"MKK7 contains three JNK-docking D-sites (D1, D2, D3) in its N-terminal domain that cooperatively promote binding to JNK1. Mutation of any single site reduces binding by 50–70%; mutation of all three reduces binding by 95%. Full-length MKK7 with D1/D2 mutations has reduced JNK1 kinase activation. D-site peptides from MKK7 inhibit JNK1 and JNK2 substrate phosphorylation selectively over ERK2.\",\n      \"method\": \"Site-directed mutagenesis, binding assay, JNK kinase assay, peptide inhibition\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — systematic mutagenesis of all three docking sites with quantitative binding and functional kinase readouts\",\n      \"pmids\": [\"16533805\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"c-FLIPL directly interacts with MKK7 in a TNFα-dependent manner and inhibits the interactions of MKK7 with MEKK1, ASK1, and TAK1. This interaction selectively suppresses the prolonged phase of JNK activation and reduces ROS accumulation in NF-κB-deficient fibroblasts.\",\n      \"method\": \"Co-immunoprecipitation, Western blot, c-Flip knockout cells, overexpression\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP in stimulated cells and knockout fibroblasts, single lab, two orthogonal approaches\",\n      \"pmids\": [\"17110930\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"In fibroblast-like synoviocytes, IL-1β-induced JNK phosphorylation and activity (measured by kinase assay), AP-1 binding, AP-1-driven transcription, and MMP-3 production are dependent on MKK7 but not MKK4. Anisomycin-induced JNK activation requires both MKK4 and MKK7.\",\n      \"method\": \"siRNA knockdown, in vitro kinase assay, EMSA, AP-1 reporter assay, ELISA\",\n      \"journal\": \"Arthritis and rheumatism\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — siRNA knockdown with multiple orthogonal functional readouts, single lab\",\n      \"pmids\": [\"16802349\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"GADD45β is a structured protein whose helices α3/α4 and loops 1/2 mediate association with MKK7. Helix α3 primarily mediates docking; loop 1 and α4-loop 2 engage the ATP-binding site of MKK7 and cause conformational changes that impede catalytic function.\",\n      \"method\": \"Structural prediction, mutagenesis, binding/inhibition assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-3 / Moderate — structural prediction with mutagenesis mapping of interaction, single lab; not a full crystal structure\",\n      \"pmids\": [\"17485467\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"GADD45β forms homodimers in solution (via helices 1 and 5) and binds tightly to MKK7; the dimerization interface (helices 1 and 5) is distinct from the MKK7-binding region. The complex may form at least a tetrameric MKK7–GADD45β:GADD45β–MKK7 unit.\",\n      \"method\": \"Size exclusion chromatography, NMR, biophysical binding assays\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR and biophysical characterization of complex, single lab\",\n      \"pmids\": [\"18343408\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The IL-1–MKK7–JNK–c-Jun signaling pathway induces EGR-1 transcription via an AP-1 site and three distal SREs. c-Jun is required for EGR-1 transcription through both elements; IL-1-inducible c-Jun recruitment to both AP-1 and SRE regions was confirmed by ChIP. EGR-1 in turn binds promoters of IL-6, IL-8, and CCL2.\",\n      \"method\": \"JNK-MKK7 fusion protein, genome-wide microarray, qPCR, reporter assay, chromatin immunoprecipitation, siRNA knockdown\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods including ChIP, microarray, reporter, and genetic KO cells\",\n      \"pmids\": [\"18281687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Rac1/MKK7/JNK signaling upregulates Atg5 in response to oncogenic H-Ras, which is critically required for autophagic cell death. Inhibition of MKK7 (but not MKK4) attenuated H-Ras(V12)-induced JNK activation; Rac1 siRNA or dominant-negative Rac1 inhibited MKK7-JNK activation and Atg5 upregulation.\",\n      \"method\": \"siRNA knockdown, dominant-negative GTPase, immunoblotting, cell death assays\",\n      \"journal\": \"Carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA and dominant-negative epistasis defining pathway, single lab\",\n      \"pmids\": [\"19783847\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Filamin A (and B, C) binds to MKK7 (specifically to the N-terminal residues 31–60 present in MKK7β and MKK7γ but not MKK7α) and also binds MKK4, connecting them in close proximity. MKK7γ (but not MKK7α) co-localizes with actin stress fibers and Filamin A. In Filamin A-deficient cells, MKK7 activation is reduced and stress-induced synergistic JNK activation is impaired.\",\n      \"method\": \"Co-immunoprecipitation, Filamin A-deficient cell line, deletion mutant analysis, confocal microscopy, kinase assay\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP, loss-of-function cell line, domain mapping, colocalization, all orthogonal\",\n      \"pmids\": [\"20156194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"MKK7 acts as a tumor suppressor coupling oncogenic/genotoxic stress to p53 stability in vivo. Tissue-specific inactivation of MKK7 in KRas(G12D)-driven lung and NeuT-driven mammary tumors markedly accelerates tumor onset. Mechanistically, MKK7 acts through JNK1 and JNK2, and this pathway directly couples oncogenic stress to p53 stability, required for cell cycle arrest.\",\n      \"method\": \"Conditional knockout (tissue-specific), tumor model, epistasis with JNK1/JNK2 knockouts, p53 stability assays\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo conditional knockout in two independent cancer models with JNK epistasis analysis\",\n      \"pmids\": [\"21317887\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"MKK7-deficient MEFs display premature senescence and G2/M arrest; MKK7 deletion in cardiomyocytes leads to heart failure under pressure overload with increased apoptosis (elevated p53, reduced MnSOD) and fibrosis (upregulated TGF-β signaling).\",\n      \"method\": \"Cardiomyocyte-specific conditional knockout, pressure overload surgery, cell death assays, Western blot\",\n      \"journal\": \"Journal of molecular and cellular cardiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional knockout with defined cardiac phenotype and mechanistic protein markers, single lab\",\n      \"pmids\": [\"21284947\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"In mkk7−/− mast cells, expression of JunB and p16INK4a are reduced and cyclin D1 is upregulated; reexpression of p16INK4a abrogates the hyperproliferative response. MKK7-regulated JNK signaling thus maintains cell cycle brakes via the p16INK4a axis.\",\n      \"method\": \"Gene knockout, flow cytometry, Western blot, adenoviral reexpression rescue\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockout with mechanistic rescue, single lab study but landmark paper\",\n      \"pmids\": [\"11560992\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Neuron-specific deletion of MKK7 impairs brain development: mice show enlarged ventricles, reduced striatum, absent axon tracts, and abnormal filamentous accumulations. MKK7 regulates axon elongation in a cell-autonomous manner and is required for contralateral axon projection by cortical neurons. JNK substrate phosphorylation (c-Jun, neurofilament heavy chain, MAP1B, doublecortin) is reduced in MKK7-deficient brain.\",\n      \"method\": \"Nestin-Cre conditional knockout, histology, electron microscopy, in utero electroporation, in vitro axon elongation assay, immunoblot of JNK substrates\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout with cell-autonomous in vitro confirmation and multiple JNK substrate readouts\",\n      \"pmids\": [\"22090513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"MKK7 mRNA localizes to the growth cone where it can be translated. MKK7 protein is phosphorylated specifically in the neurite shaft and is part of a DLK–MKK7–JNK1 module that triggers MAP1b phosphorylation to regulate microtubule bundling and neurite elongation. This local translation mechanism positions JNK signaling to microtubule regulation while uncoupling it from nuclear/transcriptional functions.\",\n      \"method\": \"Genome-wide screen for neurite-enriched mRNAs, FISH/imaging for mRNA localization, live cell imaging, immunofluorescence, pharmacological and genetic manipulation\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide screen, mRNA localization, kinase module identification with functional output, multiple orthogonal methods\",\n      \"pmids\": [\"23226105\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TIPRL (TOR signaling pathway regulator-like protein) binds to MKK7 and promotes the interaction between MKK7 and PP2Ac (protein phosphatase 2A catalytic subunit), resulting in dephosphorylation/inactivation of MKK7 and JNK, thereby conferring resistance to TRAIL-induced apoptosis in HCC cells.\",\n      \"method\": \"Co-immunoprecipitation, GST pulldown, siRNA knockdown, phospho-immunoblot, confocal microscopy\",\n      \"journal\": \"Gastroenterology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP, pulldown, knockout cells, and mechanistic phosphatase interaction identified\",\n      \"pmids\": [\"22841785\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Neuregulin/ErbB2 signaling activates Rac1, which activates MKK7 and JNK to upregulate c-Jun and downregulate Krox20 during Schwann cell dedifferentiation after nerve injury. Rac inhibition blocked MKK7 activation and c-Jun induction in sciatic nerves after axotomy.\",\n      \"method\": \"Primary Schwann cell culture, pharmacological Rac inhibition, immunoblot, ErbB2 inhibitor, microarray, axotomy model\",\n      \"journal\": \"Glia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological and in vivo inhibitor studies defining signaling pathway, single lab\",\n      \"pmids\": [\"23505039\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"GADD45β/MKK7 interaction was identified as a therapeutic target in multiple myeloma. DTP3, a D-tripeptide, disrupts the GADD45β/MKK7 complex, restores MKK7/JNK kinase activity, and kills MM cells selectively. DTP3 ablates myeloma xenografts in mice.\",\n      \"method\": \"Drug discovery/screening, co-immunoprecipitation disruption assay, JNK kinase assay, cell viability, xenograft mouse model\",\n      \"journal\": \"Cancer cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — direct complex disruption assay, kinase activity restoration, in vivo xenograft\",\n      \"pmids\": [\"25314077\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"MAVS recruits MKK7 onto mitochondria via its 3D domain upon viral infection. MKK7 subsequently phosphorylates JNK2 (not JNK1) to activate virus-induced apoptosis. Mkk7−/− cells fail to initiate virus-induced apoptosis; MAVS-MKK7-JNK2 defines a novel antiviral apoptotic pathway.\",\n      \"method\": \"Knockout cell lines (Mkk7−/−, Jnk1−/−, Jnk2−/−), co-immunoprecipitation, mitochondrial fractionation, viral challenge, Jnk2−/− mice\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic knockouts, domain mapping of MAVS-MKK7 interaction, in vivo mouse data\",\n      \"pmids\": [\"24651600\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"5Z-7-Oxozeaenol covalently binds to MAP2K7 at Cys218 (located at the end of the hinge region), not at the gatekeeper-2 cysteine, as revealed by crystal structure of the MAP2K7/5Z7O complex.\",\n      \"method\": \"X-ray crystallography\",\n      \"journal\": \"Bioorganic & medicinal chemistry letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure defining covalent binding site, single study\",\n      \"pmids\": [\"25529738\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NMR spectroscopy revealed the conformational behavior of the MKK7 regulatory domain: three docking sites (D1, D2, D3) show diverse intrinsic conformational propensities and different interaction kinetics with JNK1, though similar affinities. Crystal structure of JNK1 in complex with the second D-site of MKK7 revealed two different binding modes correlating with NMR exchange spectroscopy observations.\",\n      \"method\": \"NMR spectroscopy (atomic-resolution ensemble), X-ray crystallography\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus NMR, two orthogonal structural methods providing mechanistic insight into docking\",\n      \"pmids\": [\"25737554\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MKK7 alternative splicing during T-cell activation (skipping exon 2, mediated by CELF2 RNA-binding protein) restores a JNK-docking site disrupted in the larger isoform, enhancing JNK pathway activity (c-Jun phosphorylation, TNF-α upregulation). This splicing event is itself JNK-dependent, creating a positive feedback loop.\",\n      \"method\": \"Alternative splicing analysis, CELF2 knockdown/overexpression, phospho-immunoblot, CLIP/RNA binding assay\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mechanistic feedback loop established with genetic manipulation of CELF2 and splicing, multiple orthogonal readouts\",\n      \"pmids\": [\"26443849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Knockdown of MKK7 or ASK1 blocked DSS-induced tight junction disruption and barrier dysfunction. A Ca2+/Ask1/MKK7/JNK2/c-Src signaling cascade mediates DSS-induced tight junction disruption; JNK2-dependent Src activation leads to tyrosine phosphorylation of junctional proteins.\",\n      \"method\": \"siRNA knockdown, intracellular Ca2+ measurement, in vitro kinase assay with recombinant JNK2 and c-Src, mouse colitis model\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with in vitro kinase assay, in vivo model, single lab\",\n      \"pmids\": [\"25377781\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DUSP22 acts as a scaffold protein for the ASK1–MKK7–JNK signal transduction pathway by selectively associating with ASK1, MKK7, and JNK1/2; it increases JNK phosphorylation independently of its phosphatase activity.\",\n      \"method\": \"Co-immunoprecipitation, overexpression of phosphatase-dead mutant, JNK kinase assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP and phosphatase-dead mutant, single lab; scaffold behavior supported by dose-response analysis\",\n      \"pmids\": [\"27711255\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"KLF4 transcriptionally represses the MAP2K7 gene. In T-ALL cells with KLF4 inactivation, aberrant activation of MAP2K7 and downstream JNK and ATF2 occurs. JNK inhibitors reduce expansion of leukemia cells in patient-derived xenograft models.\",\n      \"method\": \"ChIP/gene expression, CRISPR/shRNA knockdown, xenograft model, phospho-immunoblot\",\n      \"journal\": \"Leukemia\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis by KLF4 loss-of-function with MAP2K7 activation readout, single lab\",\n      \"pmids\": [\"27872496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"SENP3-mediated deSUMOylation of MKK7 (SUMO2/3 removal) favors MKK7 binding to JNK and potentiates LPS-induced JNK phosphorylation and inflammatory cytokine production. SENP3 deficiency impairs MKK7 activity and JNK phosphorylation; ROS-dependent SENP3 accumulation after LPS drives MKK7 deSUMOylation.\",\n      \"method\": \"Conditional knockout mice, co-immunoprecipitation, in vivo and in vitro SUMO assay, phospho-immunoblot, septic shock model\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional knockout, co-IP, in vivo SUMOylation assay, multiple orthogonal methods\",\n      \"pmids\": [\"29352108\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Covalent virtual screening identified selective covalent MKK7 inhibitors. Crystal structure of lead compound bound to MKK7 confirmed the predicted binding mode. Inhibitors block JNK phosphorylation in cells, validated using MKK7 knockout cell lines, and block B-cell activation by LPS.\",\n      \"method\": \"Covalent virtual screening, X-ray crystallography, kinase selectivity panel (76 kinases), proteomics, knockout cell line validation\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure, selectivity panel, knockout validation, multiple orthogonal approaches\",\n      \"pmids\": [\"30449673\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Chemical cross-linking mass spectrometry showed that the GADD45β–MKK7 interaction largely occurs between GADD45β loop 2 (residues 103–117) and the MKK7 kinase enzymatic pocket. DTP3 disrupts this interaction by contacting MKK7 peptides 113–136 and 259–274.\",\n      \"method\": \"Chemical cross-linking mass spectrometry (CX-MS), enzymatic MS footprinting, fluorescence quenching with truncation mutants\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mass spectrometry-based structural characterization with mutagenesis validation, single lab\",\n      \"pmids\": [\"29572137\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Crystal structures of MKK7 revealed the structural basis of catalytic domain plasticity and the role of the N-terminal regulatory helix in mediating kinase activation (active-state structure). An allosteric pocket in the N-terminal lobe was identified where ibrutinib binds. Type II irreversible inhibitor binding mode and multiple novel scaffolds were identified.\",\n      \"method\": \"X-ray crystallography (comprehensive set of structures), small-molecule screening, biochemical kinase assays\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — comprehensive crystal structure set including first MAP2K active state model, multiple compounds validated\",\n      \"pmids\": [\"32783966\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"A MAP2K7 splice variant lacking exon 2 (MAP2K7Δexon2), induced by loss of MBNL1, activates JNK and promotes tumor dedifferentiation. This is the same exon-2-skipping isoform favored during T-cell activation; JNK inhibition reverses MAP2K7Δexon2-driven dedifferentiation.\",\n      \"method\": \"Alternative splicing analysis (TCGA/cancer genome), MBNL1 knockdown/overexpression, JNK inhibitor treatment, in vitro and in vivo tumor assays\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mechanistic splice-isoform validated in vitro and in vivo, single lab, corroborated by prior splicing work\",\n      \"pmids\": [\"32601196\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EEF1AKMT3 methyltransferase catalyzes monomethylation of MAP2K7 at K296, which decreases MAP2K7 phosphorylation, ubiquitination, and degradation of TP53. Loss of EEF1AKMT3 leads to MAP2K7 activation, increased tumor invasiveness and migration in gastric cancer.\",\n      \"method\": \"Mass spectrometry (substrate identification), gain/loss-of-function studies, phospho-antibody array, RNA-seq\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mass spectrometry identification of methylation site with functional gain/loss studies, single lab\",\n      \"pmids\": [\"35753528\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"MKK7 neddylation (by RanBP2ΔFG E3 ligase) reduces its basal kinase activity. RanBP2 knockdown increases MKK7 kinase activity; ectopic RanBP2ΔFG reduces it. In vitro neddylation of purified MKK7 by RanBP2ΔFG directly reduces basal kinase activity.\",\n      \"method\": \"Co-immunoprecipitation, in vitro neddylation assay, kinase activity assay, siRNA knockdown\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro neddylation reconstitution with direct kinase activity measurement, single lab\",\n      \"pmids\": [\"26364603\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RASSF7 interacts with N-Ras and MKK7 to negatively regulate JNK signaling. RASSF7 promotes the phosphorylated state of MKK7 but inhibits its ability to activate JNK. RASSF7 requires its RA domain for interaction with GTP-bound N-Ras and the anti-apoptotic effect. RASSF7 undergoes proteasomal degradation under prolonged stress, allowing MKK7/JNK death signaling to proceed.\",\n      \"method\": \"Co-immunoprecipitation, RNAi knockdown, kinase assay, domain-deletion analysis, ubiquitin-proteasome pathway assay\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, RNAi, kinase assay, domain analysis; single lab\",\n      \"pmids\": [\"21278800\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Neuron-specific MKK7 knockout mice display age-dependent motor dysfunction, axonal degeneration in the spinal cord, and accumulation of axonal transport proteins JIP1 and APP in brain and spinal cord, establishing that MKK7-JNK signaling is required for neuronal maintenance and axonal transport in adult neurons.\",\n      \"method\": \"Conditional knockout (neuron-specific), behavioral analysis, histology, immunoblot for axonal transport proteins\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional knockout with defined molecular markers, single lab\",\n      \"pmids\": [\"28779160\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MAP2K7 (MKK7) is a dual-specificity MAPK kinase that preferentially phosphorylates the Thr183 residue (within the Thr-Pro-Tyr motif) of JNK isoforms to promote their activation; it acts synergistically and non-redundantly with MKK4 (which preferentially phosphorylates Tyr185) for full JNK activation. MKK7 is the principal JNK activator downstream of proinflammatory cytokines (TNFα, IL-1), Fas, MAVS/antiviral signaling, and several MAP3Ks (DLK, MLK2, MLK3, MEKK2/3). Its N-terminal regulatory domain contains three cooperative JNK-docking D-sites and is subject to alternative splicing (exon 2 exclusion enhances JNK docking). MKK7 activity is regulated by multiple PTMs—inhibitory SUMOylation (reversed by SENP3) and neddylation (by RanBP2), and activating monomethylation at K296 by EEF1AKMT3—and by direct binding partners including GADD45β (blocks its ATP site), c-FLIPL (inhibits upstream kinase recruitment), RASSF7/N-Ras (inhibits downstream JNK activation), Filamin A (scaffolds MKK7 and MKK4), DUSP22 (scaffold for ASK1–MKK7–JNK), and TIPRL (recruits PP2Ac to dephosphorylate/inactivate MKK7). Structurally, MKK7 has been characterized by crystal structures revealing its catalytic domain plasticity, an N-terminal regulatory helix required for activation, a covalent inhibitor binding site at Cys218, and an allosteric N-lobe pocket. Physiologically, MKK7 is essential for JNK-dependent cell cycle progression (G2/M via CDC2/c-Jun), tumor suppression (coupling oncogenic stress to p53 stability), axon elongation and neuronal maintenance, cardiomyocyte stress adaptation, and negative regulation of hematopoietic cell proliferation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MAP2K7 (MKK7) is a JNK-specific dual-specificity MAP kinase kinase that constitutes the principal stress- and cytokine-activated arm of the SAPK/JNK signaling module [#0, #2]. It directly phosphorylates and activates JNK/SAPK without engaging p38 or ERK, and it does so non-redundantly with MKK4: MKK7 selectively phosphorylates the threonine (Thr183) of the JNK activation loop while MKK4 targets the tyrosine (Tyr185), so the two kinases act synergistically to achieve full dual phosphorylation and maximal JNK activity [#13, #14, #18]. Genetic disruption established that MKK7 alone is required for JNK activation by proinflammatory cytokines, whereas combined loss of MKK4 and MKK7 is needed to block stress-induced activation [#15]. MKK7 sits downstream of diverse stimuli—TNF\\u03b1, IL-1, Fas, Rac1, and antiviral MAVS signaling—and of multiple MAP3Ks including DLK, MLK2/MST, MLK3, and MEKK2/3 that preferentially feed MKK7 over MKK4 [#3, #4, #8, #9, #10, #42]. Substrate engagement is governed by an N-terminal regulatory domain bearing three cooperative JNK-docking D-sites, and alternative splicing of exon 2 modulates this docking to tune pathway output [#26, #44, #45]. MKK7 activity is constrained by an array of direct regulators: GADD45\\u03b2 binds and blocks its catalytic/ATP pocket downstream of NF-\\u03baB [#23, #29], c-FLIPL blocks upstream MAP3K recruitment [#27], TIPRL recruits PP2A to dephosphorylate it [#39], and SUMOylation (reversed by SENP3) and RanBP2-mediated neddylation impose additional negative control [#49, #55], while Filamin A and DUSP22 scaffold MKK7 with its upstream and downstream partners [#33, #47]. Through JNK\\u2013c-Jun, MKK7 couples oncogenic and genotoxic stress to p53 stability and CDC2-dependent G2/M cell cycle control, acting as a tumor suppressor and a negative regulator of hematopoietic proliferation via the p16INK4a axis [#24, #34, #16]. It is additionally essential for axon elongation, adult neuronal maintenance and axonal transport, and cardiomyocyte stress adaptation [#37, #57, #35]. Crystallographic and NMR studies have defined its catalytic-domain plasticity, an activating N-terminal regulatory helix, a covalent inhibitor site at Cys218, and an allosteric N-lobe pocket [#43, #52, #44].\",\n  \"teleology\": [\n    {\n      \"year\": 1997,\n      \"claim\": \"Establishing that a dedicated JNK-specific MAPKK existed answered whether JNK activation required a kinase distinct from the p38/JNK dual activator MKK4.\",\n      \"evidence\": \"Molecular cloning of MKK7/JNKK2 with in vitro kinase assays and Drosophila Hemipterous complementation across human and murine systems\",\n      \"pmids\": [\"9384583\", \"9312105\", \"9372971\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Site-specificity of MKK7 versus MKK4 on the JNK activation loop not yet resolved\", \"In vivo non-redundancy not yet demonstrated genetically\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Mapping MKK7 to specific receptor inputs showed it is the JNK activator selectively engaged by death-receptor and cytokine signaling rather than a generic stress kinase.\",\n      \"evidence\": \"Immune-complex kinase assays with isoform-specific antisera in Fas-stimulated cells and IL-1-stimulated rabbit liver\",\n      \"pmids\": [\"9362518\", \"9414114\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct upstream MAP3K linking receptors to MKK7 not identified\", \"Single-lab observations per stimulus\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Identifying alternatively spliced isoforms and a JNK-binding N-terminal extension explained how MKK7 docks its substrate and how basal versus inducible activity is tuned.\",\n      \"evidence\": \"Molecular cloning of six isoforms with direct binding and kinase assays plus subcellular localization\",\n      \"pmids\": [\"9891090\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological triggers selecting specific isoforms not defined\", \"Structural basis of docking not yet resolved\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Defining the MAP3K tier showed that DLK, MLK2, and MEKK2/3 preferentially activate MKK7 over MKK4, establishing pathway selectivity at the kinase-cascade level.\",\n      \"evidence\": \"In vitro reconstitution kinase assays with recombinant proteins, co-IP, and column fractionation\",\n      \"pmids\": [\"10187804\", \"10347227\", \"9516438\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Determinants of MAP3K substrate preference partly defined only for MLK2\", \"Quantitative contribution of each MAP3K in vivo unknown\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Demonstrating that MKK7 phosphorylates Thr183 while MKK4 phosphorylates Tyr185 resolved the mechanistic basis of their synergistic, non-redundant cooperation.\",\n      \"evidence\": \"In vitro kinase assays with phosphoamino-acid analysis and mass spectrometry across JNK isoforms\",\n      \"pmids\": [\"11062067\", \"10715136\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether sequential ordering of the two phosphorylations is obligatory not fully settled\", \"Isoform-specific kinetics in cells not measured\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Genetic knockout established the in vivo division of labor: MKK7 is essential for cytokine-induced JNK activation, while stress activation requires both MKK7 and MKK4.\",\n      \"evidence\": \"Targeted Mkk7 and Mkk4/Mkk7 gene disruption in mice with phospho-specific JNK readouts\",\n      \"pmids\": [\"11390361\", \"11418587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cell-type-specific differences in input dependence not fully mapped\", \"Isoform-specific contributions not separable by knockout\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Conditional deletion revealed MKK7 as a negative regulator of hematopoietic proliferation acting through the p16INK4a cell-cycle brake.\",\n      \"evidence\": \"Conditional gene targeting in thymocytes, B cells, and mast cells with p16INK4a reexpression rescue\",\n      \"pmids\": [\"11560992\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct molecular link between JNK signaling and p16INK4a induction not defined\", \"Whether effect generalizes beyond hematopoietic lineages unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Linking MKK7-JNK-c-Jun to CDC2 placed the pathway directly in control of G2/M progression and senescence, framing its cell-cycle function.\",\n      \"evidence\": \"Gene knockout in MEFs with cell cycle analysis and c-Jun mutant epistasis\",\n      \"pmids\": [\"15039780\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which c-Jun regulates CDC2 not detailed\", \"Connection to p16/p53 arms not integrated\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identifying GADD45\\u03b2 as a direct catalytic inhibitor explained how NF-\\u03baB suppresses JNK and protects against TNF\\u03b1 cytotoxicity.\",\n      \"evidence\": \"Unbiased screen, direct binding, kinase inhibition, and disrupting-peptide rescue; later refined by structural mapping of the ATP-pocket-engaging interface\",\n      \"pmids\": [\"14743220\", \"17485467\", \"18343408\", \"29572137\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and full structure of the complex defined only by prediction/NMR, not crystallography\", \"Regulation of GADD45\\u03b2 abundance in vivo incompletely defined\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Systematic mapping of three cooperative N-terminal D-sites established the structural logic of MKK7\\u2013JNK substrate docking.\",\n      \"evidence\": \"Site-directed mutagenesis of all three docking sites with quantitative binding and kinase assays, later extended by NMR and crystallography\",\n      \"pmids\": [\"16533805\", \"25737554\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of distinct docking-site binding modes not fully resolved\", \"Regulation of docking by phosphorylation/PTMs not addressed\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Discovery of scaffolds and additional binding-partner regulators (Filamin A, RASSF7/N-Ras) showed how MKK7 is spatially organized and its output set by competing inputs.\",\n      \"evidence\": \"Co-IP, loss-of-function cell lines, domain mapping, and colocalization\",\n      \"pmids\": [\"20156194\", \"21278800\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"RASSF7 mechanism (phospho-MKK7 yet inhibited JNK output) not mechanistically explained\", \"Scaffold contributions in vivo not tested genetically\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"In vivo tumor models established MKK7 as a tumor suppressor coupling oncogenic stress to p53 stability through JNK1/2.\",\n      \"evidence\": \"Tissue-specific conditional knockout in KRas- and NeuT-driven tumor models with JNK1/JNK2 epistasis and p53 stability assays\",\n      \"pmids\": [\"21317887\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular step connecting JNK to p53 stabilization not defined\", \"Relevance to human tumors with MAP2K7 alterations not established here\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Conditional deletion in neurons and cardiomyocytes defined tissue-specific physiological roles in axon elongation, neuronal maintenance, and cardiac stress adaptation.\",\n      \"evidence\": \"Nestin-Cre and cardiomyocyte-specific knockouts with histology, axon assays, pressure-overload surgery, and JNK substrate immunoblots\",\n      \"pmids\": [\"22090513\", \"21284947\", \"28779160\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which JNK substrates dominate each phenotype not fully dissected\", \"Cardiac and neuronal effects characterized in single labs\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Local mRNA translation at the growth cone and a DLK\\u2013MKK7\\u2013JNK1 module showed MKK7 can act on cytoskeletal substrates (MAP1B) spatially uncoupled from nuclear transcription.\",\n      \"evidence\": \"Genome-wide neurite mRNA screen, mRNA localization imaging, and kinase-module manipulation\",\n      \"pmids\": [\"23226105\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of localized translation by upstream signals not defined\", \"Generality beyond neurons unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identifying MAVS-mediated mitochondrial recruitment and selective JNK2 phosphorylation defined an antiviral apoptotic function for MKK7.\",\n      \"evidence\": \"Multiple genetic knockouts, MAVS domain mapping, mitochondrial fractionation, and viral challenge in mice\",\n      \"pmids\": [\"24651600\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Basis for JNK2 versus JNK1 selectivity not mechanistically resolved\", \"Interplay with other antiviral pathways not addressed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Druggability was established by targeting the GADD45\\u03b2/MKK7 interface and by covalent occupancy of Cys218, opening therapeutic and chemical-biology avenues.\",\n      \"evidence\": \"DTP3 complex-disruption assays with xenografts, plus crystal structures of covalent inhibitor complexes and selective inhibitor development validated in knockout cells\",\n      \"pmids\": [\"25314077\", \"25529738\", \"30449673\", \"29572137\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Clinical translation of these agents not addressed in the corpus\", \"Off-target/selectivity profiles defined only for specific scaffolds\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"A layered PTM code (SUMOylation/SENP3, RanBP2 neddylation, EEF1AKMT3 methylation) and CELF2/MBNL1-controlled exon-2 splicing revealed how MKK7 activity is set post-translationally and post-transcriptionally.\",\n      \"evidence\": \"Conditional knockouts, in vitro neddylation/SUMO assays, mass-spectrometric methylation-site identification, and splicing manipulation\",\n      \"pmids\": [\"26443849\", \"29352108\", \"26364603\", \"35753528\", \"32601196\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Hierarchy and crosstalk among the different PTMs not integrated\", \"In vivo importance of each modification context-limited\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Comprehensive crystal structures defined catalytic-domain plasticity, an activating N-terminal regulatory helix, and an allosteric N-lobe pocket, providing a structural framework for MKK7 activation and inhibition.\",\n      \"evidence\": \"Comprehensive X-ray crystallography including an active-state model plus small-molecule screening and kinase assays\",\n      \"pmids\": [\"32783966\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of full-length MKK7 with regulatory domain and docked JNK not solved\", \"Conformational dynamics during the activation cycle not fully captured\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple negative regulators, PTMs, scaffolds, and isoforms are integrated to set MKK7 output in a given cell type and stimulus, and how this is therapeutically exploitable beyond GADD45\\u03b2 disruption, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified quantitative model of MKK7 regulation across inputs\", \"Context-specific dominance of each regulator unknown\", \"In vivo consequences of combined PTM perturbation untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 13, 14]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 13, 18]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [23, 29, 43]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [7, 21]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [7, 11]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [42]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [33, 38]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 13, 15]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [4, 15, 28, 49]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [24, 34]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [23, 42, 56]}\n    ],\n    \"complexes\": [\n      \"MKK4\\u2013MKK7\\u2013JNK complex\",\n      \"ASK1\\u2013MKK7\\u2013JNK (DUSP22-scaffolded)\",\n      \"GADD45\\u03b2\\u2013MKK7 inhibitory complex\"\n    ],\n    \"partners\": [\n      \"MAPK8\",\n      \"MAP2K4\",\n      \"GADD45B\",\n      \"MAP3K12\",\n      \"FLNA\",\n      \"TIPRL\",\n      \"DUSP22\",\n      \"RASSF7\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":9,"faith_total":9,"faith_pct":100.0}}