{"gene":"DUSP7","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":1998,"finding":"PYST2/DUSP7 protein is predominantly cytosolic when expressed in COS-1 cells. It shows substrate selectivity for ERK2 (p42 MAP kinase) both in vitro and in vivo, with much reduced activity toward stress-activated kinases JNK-1 and p38/RK. PYST2 binds p42 MAP kinase in vivo, and both MAP and SAP kinases can cause catalytic activation of PYST2 phosphatase activity in vitro.","method":"Subcellular fractionation/localization in COS-1 cells, in vitro phosphatase assays, in vivo substrate selectivity assays, in vivo co-binding experiments","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro enzymatic assays combined with in vivo substrate selectivity and binding experiments, multiple orthogonal methods in a single focused study","pmids":["9788880"],"is_preprint":false},{"year":2004,"finding":"Overexpression of Pyst2-L in HEK293 cells reduced basal phospho-ERK1/2 levels. Pull-down experiments showed that recombinant Pyst2-L binds not only phospho-ERK1/2 but also phospho-JNK, indicating the phosphatase is not exclusively selective for ERK2.","method":"Overexpression in HEK293 cells with Western blot for phospho-ERK1/2; pull-down assay with recombinant Pyst2-L for phospho-JNK","journal":"Immunology letters","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal pull-down and overexpression with Western blot, single lab, two orthogonal methods but limited controls","pmids":["15081539"],"is_preprint":false},{"year":2004,"finding":"The PYST2 locus encodes two alternatively transcribed and translated isoforms: PYST2-L (containing the phosphatase catalytic domain, PCD) and PYST2-S (lacking any known PCD). Both transcripts are translated into protein. PYST2-S may act as a negative regulator of PYST2-L.","method":"Northern blot, RT-PCR, sequencing, Western blot, bioinformatic domain analysis","journal":"Genes, chromosomes & cancer","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Northern blot and Western blot confirm two isoforms, but negative regulatory role of PYST2-S is speculative and not experimentally confirmed","pmids":["14603440"],"is_preprint":false},{"year":2005,"finding":"Pyst2-L expression is reduced in G2-phase-synchronized human K562 cells, consistent with homologous genes (yeast Yvh1p and Xenopus X17c) that regulate cell cycle. Cells in highly crowded cultures express high levels of Pyst2-L phosphatase, implicating it in crowding-induced signaling.","method":"Cell cycle synchronization with alpha-factor, Northern blot/expression analysis in synchronized K562 cells; comparative genomic analysis; expression measurement in crowded vs. non-crowded cultures","journal":"Immunology letters","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single expression-based method per observation, no direct functional rescue or mechanistic pathway placement","pmids":["16386315"],"is_preprint":false},{"year":2016,"finding":"DUSP7 is required for meiotic resumption in mouse oocytes. DUSP7-depleted oocytes fail to resume meiosis or do so with significant delay due to Cdk1/CycB activity dropping below the critical threshold for nuclear envelope breakdown. DUSP7 drives meiotic resumption by dephosphorylating and inactivating cPKC isoforms. Additionally, DUSP7 is required for proper chromosome alignment; DUSP7-depleted oocytes entering meiosis show severe chromosome alignment defects and premature anaphase progression.","method":"RNAi-mediated depletion in mouse oocytes, live cell imaging, Cdk1/CycB activity assays, cPKC phosphorylation analysis","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — loss-of-function with multiple defined phenotypic readouts, kinase activity measurements, and substrate (cPKC) identification in a single rigorous study","pmids":["27783954"],"is_preprint":false},{"year":2021,"finding":"During mitosis, DUSP7 binds ERK2 and regulates the abundance of active phospho-ERK2 through its phosphatase activity. Overexpression of catalytically active DUSP7 (but not catalytically inactive mutants) decreases phospho-ERK2 levels and causes mitotic chromosome misalignment. Knockdown of DUSP7 also leads to defective chromosome congression and prolonged mitosis. Proper chromosome alignment requires MEK-mediated phosphorylation and DUSP7-mediated dephosphorylation to maintain appropriate levels of active phospho-ERK2.","method":"Co-immunoprecipitation of DUSP7 with ERK2 during mitosis, catalytic mutant overexpression, siRNA knockdown, immunofluorescence for chromosome alignment, phospho-ERK2 Western blot, chemical inhibition of ERK2 and MEK","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — Co-IP, catalytic mutant validation, RNAi, chemical inhibition, and direct phospho-substrate measurement; multiple orthogonal methods in one study","pmids":["33865857"],"is_preprint":false},{"year":2021,"finding":"DUSP7 overexpression in cervical cancer SIHA cells decreases HRAS and phospho-ERK1/2 levels, suppresses invasion, migration, proliferation in vitro, and tumor formation in vivo, indicating DUSP7 inhibits cancer progression by inactivating the RAS/ERK pathway.","method":"DUSP7 overexpression and shRNA knockdown in SIHA cells, Western blot for HRAS and p-ERK1/2, in vitro invasion/migration/proliferation assays, in vivo xenograft tumor formation","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — loss- and gain-of-function with pathway readouts, single lab, multiple assays but no direct biochemical reconstitution","pmids":["34435746"],"is_preprint":false},{"year":2022,"finding":"FOSL1 transcription factor positively regulates DUSP7 transcription in doxorubicin-resistant breast cancer cells. DUSP7 in turn mediates dephosphorylation of PEA15 (proliferation and apoptosis adaptor protein 15), promoting drug resistance through this FOSL1/DUSP7/PEA15 pathway.","method":"qRT-PCR for FOSL1 and DUSP7, transcriptional reporter/ChIP-type assays, Western blot for phospho-PEA15, in vitro and in vivo drug resistance assays","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — identification of novel substrate PEA15 and upstream transcriptional regulator FOSL1, single lab with multiple in vitro and in vivo methods","pmids":["34907034"],"is_preprint":false},{"year":2020,"finding":"DUSP7 physically interacts with TAK1 (TGF-β-activated kinase 1) in hepatocytes. DUSP7 knockout in mice promotes TAK1 activation, leading to increased lipid deposition, inflammatory response (via NF-κB and MAPK pathways), and ROS production in HFD-fed conditions. DUSP7 thus acts as a negative regulator of TAK1-driven NAFLD pathogenesis.","method":"Co-immunoprecipitation (physical interaction with TAK1), DUSP7 knockout mouse model, HFD-feeding experiments, Western blot for phospho-TAK1 and downstream pathway components, metabolic phenotyping","journal":"Free radical biology & medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for TAK1 interaction and genetic KO with defined pathway readouts, single lab","pmids":["32311490"],"is_preprint":false}],"current_model":"DUSP7 (PYST2/MKP-X) is a cytosolic dual-specificity MAP kinase phosphatase that preferentially dephosphorylates and inactivates ERK2 (p42), with catalytic activity allosterically stimulated by ERK2 binding; it also dephosphorylates cPKC isoforms to drive meiotic resumption in oocytes and dephosphorylates PEA15 in cancer cells, and physically interacts with TAK1 to suppress inflammatory signaling, with balanced MEK-driven phosphorylation and DUSP7-driven dephosphorylation of ERK2 being required for proper chromosome alignment during both mitosis and meiosis."},"narrative":{"mechanistic_narrative":"DUSP7 (PYST2/MKP-X) is a predominantly cytosolic dual-specificity MAP kinase phosphatase that controls the amplitude of MAPK signaling by dephosphorylating and inactivating ERK2, toward which it shows substrate selectivity and to which it binds, with its catalytic activity allosterically stimulated upon MAP kinase binding [PMID:9788880]. Through this ERK2-directed activity DUSP7 functions as a tunable counterweight to MEK during cell division: it binds active phospho-ERK2 during mitosis, and the balance between MEK-driven phosphorylation and DUSP7-driven dephosphorylation must be maintained for proper chromosome congression, as both catalytically active DUSP7 overexpression and DUSP7 depletion cause chromosome misalignment and prolonged mitosis [PMID:33865857]. The same requirement extends to meiosis, where DUSP7 drives meiotic resumption in mouse oocytes by dephosphorylating and inactivating cPKC isoforms to sustain Cdk1/CycB activity above the threshold for nuclear envelope breakdown, and is again needed for correct chromosome alignment [PMID:27783954]. Beyond ERK2, DUSP7 acts on additional substrates and partners in disease contexts: it suppresses the RAS/ERK pathway to restrain cancer cell invasion, migration, proliferation and tumor formation [PMID:34435746], dephosphorylates the adaptor PEA15 downstream of FOSL1 to promote drug resistance in breast cancer [PMID:34907034], and physically interacts with TAK1 in hepatocytes to negatively regulate inflammatory and lipogenic signaling in NAFLD [PMID:32311490]. The DUSP7 locus encodes a catalytic isoform (PYST2-L) and a shorter isoform lacking the phosphatase domain (PYST2-S) [PMID:14603440].","teleology":[{"year":1998,"claim":"Established DUSP7's core biochemical identity by defining it as a cytosolic dual-specificity phosphatase selective for ERK2 whose activity is switched on by MAP kinase binding.","evidence":"Subcellular fractionation, in vitro phosphatase assays, and in vivo substrate selectivity/binding in COS-1 cells","pmids":["9788880"],"confidence":"High","gaps":["Structural basis of allosteric activation by ERK2 not resolved","Physiological signaling context for ERK2 inactivation not yet addressed"]},{"year":2004,"claim":"Refined the substrate specificity question by showing recombinant DUSP7 can also bind phospho-JNK, indicating ERK2 selectivity is not absolute.","evidence":"Overexpression with phospho-ERK1/2 Western blot and recombinant pull-down for phospho-JNK in HEK293 cells","pmids":["15081539"],"confidence":"Medium","gaps":["JNK binding shown by pull-down without functional dephosphorylation in cells","Limited controls for binding specificity"]},{"year":2004,"claim":"Defined the gene architecture by identifying a catalytic isoform (PYST2-L) and a catalytically deficient short isoform (PYST2-S), raising the possibility of intramolecular regulation.","evidence":"Northern blot, RT-PCR, sequencing, Western blot, and domain analysis","pmids":["14603440"],"confidence":"Medium","gaps":["Negative-regulatory role of PYST2-S inferred but not experimentally demonstrated","Relative abundance and tissue distribution of isoforms unknown"]},{"year":2005,"claim":"Linked DUSP7 expression to cell-cycle phase and culture density, hinting at a cell-cycle regulatory function.","evidence":"Cell cycle synchronization and expression analysis in K562 cells plus crowding comparisons","pmids":["16386315"],"confidence":"Low","gaps":["Correlative expression only, no functional rescue or pathway placement","Mechanism connecting crowding to DUSP7 expression unknown"]},{"year":2016,"claim":"Demonstrated a physiological requirement for DUSP7 in meiosis, identifying cPKC isoforms as substrates whose dephosphorylation drives meiotic resumption and proper chromosome alignment.","evidence":"RNAi depletion in mouse oocytes with live imaging, Cdk1/CycB activity assays, and cPKC phosphorylation analysis","pmids":["27783954"],"confidence":"High","gaps":["Direct enzyme-substrate dephosphorylation of cPKC not reconstituted in vitro","How cPKC links to the Cdk1/CycB threshold not fully mapped"]},{"year":2020,"claim":"Extended DUSP7's regulatory reach to inflammatory metabolism by showing it physically restrains TAK1 to limit NAFLD pathogenesis.","evidence":"Co-IP for TAK1 interaction and DUSP7 knockout mice under HFD with pathway and metabolic readouts","pmids":["32311490"],"confidence":"Medium","gaps":["Whether DUSP7 dephosphorylates TAK1 or acts as a scaffold not distinguished","Single Co-IP without reciprocal biochemical validation of mechanism"]},{"year":2021,"claim":"Resolved the mitotic function of ERK2 dephosphorylation, establishing that a MEK/DUSP7 balance setting phospho-ERK2 levels governs chromosome congression.","evidence":"Co-IP of DUSP7 with ERK2 in mitosis, catalytic mutant overexpression, siRNA, chemical MEK/ERK inhibition, and chromosome alignment imaging","pmids":["33865857"],"confidence":"High","gaps":["ERK2 substrates relevant to chromosome alignment not identified","Spatial regulation of DUSP7 activity at the spindle/kinetochore unknown"]},{"year":2021,"claim":"Positioned DUSP7 as a tumor suppressor in cervical cancer acting through inactivation of the RAS/ERK pathway.","evidence":"Overexpression and shRNA knockdown in SIHA cells with HRAS/p-ERK Western blots, in vitro phenotypic assays, and xenografts","pmids":["34435746"],"confidence":"Medium","gaps":["Mechanism by which DUSP7 lowers HRAS not defined","No direct biochemical reconstitution of pathway suppression"]},{"year":2022,"claim":"Identified a FOSL1/DUSP7/PEA15 axis in which transcriptionally induced DUSP7 dephosphorylates PEA15 to promote chemoresistance, adding a new substrate and an upstream regulator.","evidence":"qRT-PCR, transcriptional/ChIP-type assays, phospho-PEA15 Western blot, and in vitro/in vivo drug resistance assays in breast cancer cells","pmids":["34907034"],"confidence":"Medium","gaps":["Direct DUSP7-PEA15 dephosphorylation not reconstituted in vitro","How PEA15 dephosphorylation confers resistance mechanistically unresolved"]},{"year":null,"claim":"How DUSP7's substrate choice (ERK2, cPKC, PEA15, TAK1) is partitioned across cell types, cell-cycle phases, and disease contexts, and the structural basis of its allosteric activation, remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of substrate engagement or activation","No unified framework reconciling ERK2-selective and multi-substrate activities","Determinants directing DUSP7 to distinct substrates in different tissues unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,4,5,7]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,5]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,5,6]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4,5]}],"complexes":[],"partners":["MAPK1","TAK1","PEA15"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q16829","full_name":"Dual specificity protein phosphatase 7","aliases":["Dual specificity protein phosphatase PYST2"],"length_aa":419,"mass_kda":45.0,"function":"Dual specificity protein phosphatase (PubMed:9788880). Shows high activity towards MAPK1/ERK2 (PubMed:9788880). Also has lower activity towards MAPK14 and MAPK8 (PubMed:9788880). In arrested oocytes, plays a role in meiotic resumption (By similarity). Promotes nuclear envelope breakdown and activation of the CDK1/Cyclin-B complex in oocytes, probably by dephosphorylating and inactivating the conventional protein kinase C (cPKC) isozyme PRKCB (By similarity). May also inactivate PRKCA and/or PRKCG (By similarity). Also important in oocytes for normal chromosome alignment on the metaphase plate and progression to anaphase, where it might regulate activity of the spindle-assembly checkpoint (SAC) complex (By similarity)","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q16829/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DUSP7","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DUSP7","total_profiled":1310},"omim":[{"mim_id":"602749","title":"DUAL-SPECIFICITY PHOSPHATASE 7; DUSP7","url":"https://www.omim.org/entry/602749"},{"mim_id":"602748","title":"DUAL-SPECIFICITY PHOSPHATASE 6; DUSP6","url":"https://www.omim.org/entry/602748"},{"mim_id":"602747","title":"DUAL-SPECIFICITY PHOSPHATASE 4; DUSP4","url":"https://www.omim.org/entry/602747"},{"mim_id":"602209","title":"RAS-RESPONSIVE ELEMENT BINDING PROTEIN 1; RREB1","url":"https://www.omim.org/entry/602209"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DUSP7"},"hgnc":{"alias_symbol":["MKP-X","PYST2"],"prev_symbol":[]},"alphafold":{"accession":"Q16829","domains":[{"cath_id":"3.40.250.10","chopping":"55-187","consensus_level":"high","plddt":83.6798,"start":55,"end":187},{"cath_id":"3.90.190.10","chopping":"247-386","consensus_level":"high","plddt":94.5321,"start":247,"end":386}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q16829","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q16829-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q16829-F1-predicted_aligned_error_v6.png","plddt_mean":74.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DUSP7","jax_strain_url":"https://www.jax.org/strain/search?query=DUSP7"},"sequence":{"accession":"Q16829","fasta_url":"https://rest.uniprot.org/uniprotkb/Q16829.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q16829/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q16829"}},"corpus_meta":[{"pmid":"9788880","id":"PMC_9788880","title":"Isolation of the human genes encoding the pyst1 and Pyst2 phosphatases: characterisation of Pyst2 as a cytosolic dual-specificity MAP kinase phosphatase and its catalytic activation by both MAP and SAP kinases.","date":"1998","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/9788880","citation_count":110,"is_preprint":false},{"pmid":"29100300","id":"PMC_29100300","title":"Long non-coding RNA MIAT promotes breast cancer progression and functions as ceRNA to regulate DUSP7 expression by sponging miR-155-5p.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29100300","citation_count":105,"is_preprint":false},{"pmid":"32273769","id":"PMC_32273769","title":"Circ-ABCB10 Contributes to Paclitaxel Resistance in Breast Cancer Through Let-7a-5p/DUSP7 Axis.","date":"2020","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/32273769","citation_count":81,"is_preprint":false},{"pmid":"18058922","id":"PMC_18058922","title":"Expression of ERK signaling inhibitors Dusp6, Dusp7, and Dusp9 during mouse ear development.","date":"2008","source":"Developmental dynamics : an official publication of the American Association of Anatomists","url":"https://pubmed.ncbi.nlm.nih.gov/18058922","citation_count":33,"is_preprint":false},{"pmid":"12969791","id":"PMC_12969791","title":"Overexpression of the dual-specificity MAPK phosphatase PYST2 in acute leukemia.","date":"2003","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/12969791","citation_count":27,"is_preprint":false},{"pmid":"14576828","id":"PMC_14576828","title":"Dual-specificity phosphatase Pyst2-L is constitutively highly expressed in myeloid leukemia and other malignant cells.","date":"2003","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/14576828","citation_count":27,"is_preprint":false},{"pmid":"32311490","id":"PMC_32311490","title":"Targeting DUSP7 signaling alleviates hepatic steatosis, inflammation and oxidative stress in high fat diet (HFD)-fed mice via suppression of TAK1.","date":"2020","source":"Free radical biology & medicine","url":"https://pubmed.ncbi.nlm.nih.gov/32311490","citation_count":26,"is_preprint":false},{"pmid":"9205128","id":"PMC_9205128","title":"Chromosomal localization of three human dual specificity phosphatase genes (DUSP4, DUSP6, and DUSP7).","date":"1997","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/9205128","citation_count":25,"is_preprint":false},{"pmid":"34907034","id":"PMC_34907034","title":"Transcription Factor FOSL1 Enhances Drug Resistance of Breast Cancer through DUSP7-Mediated Dephosphorylation of PEA15.","date":"2022","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/34907034","citation_count":23,"is_preprint":false},{"pmid":"27783954","id":"PMC_27783954","title":"The Phosphatase Dusp7 Drives Meiotic Resumption and Chromosome Alignment in Mouse Oocytes.","date":"2016","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/27783954","citation_count":18,"is_preprint":false},{"pmid":"33865857","id":"PMC_33865857","title":"DUSP7 regulates the activity of ERK2 to promote proper chromosome alignment during cell division.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33865857","citation_count":12,"is_preprint":false},{"pmid":"34435746","id":"PMC_34435746","title":"DUSP7 inhibits cervical cancer progression by inactivating the RAS pathway.","date":"2021","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34435746","citation_count":10,"is_preprint":false},{"pmid":"15081539","id":"PMC_15081539","title":"Does the dual-specificity MAPK phosphatase Pyst2-L lead a monogamous relationship with the Erk2 protein?","date":"2004","source":"Immunology letters","url":"https://pubmed.ncbi.nlm.nih.gov/15081539","citation_count":10,"is_preprint":false},{"pmid":"28253537","id":"PMC_28253537","title":"mRNA profiling identifies low levels of phosphatases dual‐specific phosphatase‐7 (DUSP7) and cell division cycle‐25B (CDC25B) in patients with early arthritis.","date":"2017","source":"Clinical and experimental immunology","url":"https://pubmed.ncbi.nlm.nih.gov/28253537","citation_count":10,"is_preprint":false},{"pmid":"14603440","id":"PMC_14603440","title":"Characterization of the dual-specificity phosphatase PYST2 and its transcripts.","date":"2004","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/14603440","citation_count":3,"is_preprint":false},{"pmid":"16386315","id":"PMC_16386315","title":"The Pyst2-L phosphatase is involved in cell-crowding.","date":"2005","source":"Immunology letters","url":"https://pubmed.ncbi.nlm.nih.gov/16386315","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9879,"output_tokens":2581,"usd":0.034176,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9778,"output_tokens":3271,"usd":0.065333,"stage2_stop_reason":"end_turn"},"total_usd":0.099509,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1998,\n      \"finding\": \"PYST2/DUSP7 protein is predominantly cytosolic when expressed in COS-1 cells. It shows substrate selectivity for ERK2 (p42 MAP kinase) both in vitro and in vivo, with much reduced activity toward stress-activated kinases JNK-1 and p38/RK. PYST2 binds p42 MAP kinase in vivo, and both MAP and SAP kinases can cause catalytic activation of PYST2 phosphatase activity in vitro.\",\n      \"method\": \"Subcellular fractionation/localization in COS-1 cells, in vitro phosphatase assays, in vivo substrate selectivity assays, in vivo co-binding experiments\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro enzymatic assays combined with in vivo substrate selectivity and binding experiments, multiple orthogonal methods in a single focused study\",\n      \"pmids\": [\"9788880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Overexpression of Pyst2-L in HEK293 cells reduced basal phospho-ERK1/2 levels. Pull-down experiments showed that recombinant Pyst2-L binds not only phospho-ERK1/2 but also phospho-JNK, indicating the phosphatase is not exclusively selective for ERK2.\",\n      \"method\": \"Overexpression in HEK293 cells with Western blot for phospho-ERK1/2; pull-down assay with recombinant Pyst2-L for phospho-JNK\",\n      \"journal\": \"Immunology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal pull-down and overexpression with Western blot, single lab, two orthogonal methods but limited controls\",\n      \"pmids\": [\"15081539\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The PYST2 locus encodes two alternatively transcribed and translated isoforms: PYST2-L (containing the phosphatase catalytic domain, PCD) and PYST2-S (lacking any known PCD). Both transcripts are translated into protein. PYST2-S may act as a negative regulator of PYST2-L.\",\n      \"method\": \"Northern blot, RT-PCR, sequencing, Western blot, bioinformatic domain analysis\",\n      \"journal\": \"Genes, chromosomes & cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Northern blot and Western blot confirm two isoforms, but negative regulatory role of PYST2-S is speculative and not experimentally confirmed\",\n      \"pmids\": [\"14603440\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Pyst2-L expression is reduced in G2-phase-synchronized human K562 cells, consistent with homologous genes (yeast Yvh1p and Xenopus X17c) that regulate cell cycle. Cells in highly crowded cultures express high levels of Pyst2-L phosphatase, implicating it in crowding-induced signaling.\",\n      \"method\": \"Cell cycle synchronization with alpha-factor, Northern blot/expression analysis in synchronized K562 cells; comparative genomic analysis; expression measurement in crowded vs. non-crowded cultures\",\n      \"journal\": \"Immunology letters\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single expression-based method per observation, no direct functional rescue or mechanistic pathway placement\",\n      \"pmids\": [\"16386315\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DUSP7 is required for meiotic resumption in mouse oocytes. DUSP7-depleted oocytes fail to resume meiosis or do so with significant delay due to Cdk1/CycB activity dropping below the critical threshold for nuclear envelope breakdown. DUSP7 drives meiotic resumption by dephosphorylating and inactivating cPKC isoforms. Additionally, DUSP7 is required for proper chromosome alignment; DUSP7-depleted oocytes entering meiosis show severe chromosome alignment defects and premature anaphase progression.\",\n      \"method\": \"RNAi-mediated depletion in mouse oocytes, live cell imaging, Cdk1/CycB activity assays, cPKC phosphorylation analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — loss-of-function with multiple defined phenotypic readouts, kinase activity measurements, and substrate (cPKC) identification in a single rigorous study\",\n      \"pmids\": [\"27783954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"During mitosis, DUSP7 binds ERK2 and regulates the abundance of active phospho-ERK2 through its phosphatase activity. Overexpression of catalytically active DUSP7 (but not catalytically inactive mutants) decreases phospho-ERK2 levels and causes mitotic chromosome misalignment. Knockdown of DUSP7 also leads to defective chromosome congression and prolonged mitosis. Proper chromosome alignment requires MEK-mediated phosphorylation and DUSP7-mediated dephosphorylation to maintain appropriate levels of active phospho-ERK2.\",\n      \"method\": \"Co-immunoprecipitation of DUSP7 with ERK2 during mitosis, catalytic mutant overexpression, siRNA knockdown, immunofluorescence for chromosome alignment, phospho-ERK2 Western blot, chemical inhibition of ERK2 and MEK\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — Co-IP, catalytic mutant validation, RNAi, chemical inhibition, and direct phospho-substrate measurement; multiple orthogonal methods in one study\",\n      \"pmids\": [\"33865857\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"DUSP7 overexpression in cervical cancer SIHA cells decreases HRAS and phospho-ERK1/2 levels, suppresses invasion, migration, proliferation in vitro, and tumor formation in vivo, indicating DUSP7 inhibits cancer progression by inactivating the RAS/ERK pathway.\",\n      \"method\": \"DUSP7 overexpression and shRNA knockdown in SIHA cells, Western blot for HRAS and p-ERK1/2, in vitro invasion/migration/proliferation assays, in vivo xenograft tumor formation\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — loss- and gain-of-function with pathway readouts, single lab, multiple assays but no direct biochemical reconstitution\",\n      \"pmids\": [\"34435746\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FOSL1 transcription factor positively regulates DUSP7 transcription in doxorubicin-resistant breast cancer cells. DUSP7 in turn mediates dephosphorylation of PEA15 (proliferation and apoptosis adaptor protein 15), promoting drug resistance through this FOSL1/DUSP7/PEA15 pathway.\",\n      \"method\": \"qRT-PCR for FOSL1 and DUSP7, transcriptional reporter/ChIP-type assays, Western blot for phospho-PEA15, in vitro and in vivo drug resistance assays\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — identification of novel substrate PEA15 and upstream transcriptional regulator FOSL1, single lab with multiple in vitro and in vivo methods\",\n      \"pmids\": [\"34907034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DUSP7 physically interacts with TAK1 (TGF-β-activated kinase 1) in hepatocytes. DUSP7 knockout in mice promotes TAK1 activation, leading to increased lipid deposition, inflammatory response (via NF-κB and MAPK pathways), and ROS production in HFD-fed conditions. DUSP7 thus acts as a negative regulator of TAK1-driven NAFLD pathogenesis.\",\n      \"method\": \"Co-immunoprecipitation (physical interaction with TAK1), DUSP7 knockout mouse model, HFD-feeding experiments, Western blot for phospho-TAK1 and downstream pathway components, metabolic phenotyping\",\n      \"journal\": \"Free radical biology & medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for TAK1 interaction and genetic KO with defined pathway readouts, single lab\",\n      \"pmids\": [\"32311490\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DUSP7 (PYST2/MKP-X) is a cytosolic dual-specificity MAP kinase phosphatase that preferentially dephosphorylates and inactivates ERK2 (p42), with catalytic activity allosterically stimulated by ERK2 binding; it also dephosphorylates cPKC isoforms to drive meiotic resumption in oocytes and dephosphorylates PEA15 in cancer cells, and physically interacts with TAK1 to suppress inflammatory signaling, with balanced MEK-driven phosphorylation and DUSP7-driven dephosphorylation of ERK2 being required for proper chromosome alignment during both mitosis and meiosis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DUSP7 (PYST2/MKP-X) is a predominantly cytosolic dual-specificity MAP kinase phosphatase that controls the amplitude of MAPK signaling by dephosphorylating and inactivating ERK2, toward which it shows substrate selectivity and to which it binds, with its catalytic activity allosterically stimulated upon MAP kinase binding [#0]. Through this ERK2-directed activity DUSP7 functions as a tunable counterweight to MEK during cell division: it binds active phospho-ERK2 during mitosis, and the balance between MEK-driven phosphorylation and DUSP7-driven dephosphorylation must be maintained for proper chromosome congression, as both catalytically active DUSP7 overexpression and DUSP7 depletion cause chromosome misalignment and prolonged mitosis [#5]. The same requirement extends to meiosis, where DUSP7 drives meiotic resumption in mouse oocytes by dephosphorylating and inactivating cPKC isoforms to sustain Cdk1/CycB activity above the threshold for nuclear envelope breakdown, and is again needed for correct chromosome alignment [#4]. Beyond ERK2, DUSP7 acts on additional substrates and partners in disease contexts: it suppresses the RAS/ERK pathway to restrain cancer cell invasion, migration, proliferation and tumor formation [#6], dephosphorylates the adaptor PEA15 downstream of FOSL1 to promote drug resistance in breast cancer [#7], and physically interacts with TAK1 in hepatocytes to negatively regulate inflammatory and lipogenic signaling in NAFLD [#8]. The DUSP7 locus encodes a catalytic isoform (PYST2-L) and a shorter isoform lacking the phosphatase domain (PYST2-S) [#2].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established DUSP7's core biochemical identity by defining it as a cytosolic dual-specificity phosphatase selective for ERK2 whose activity is switched on by MAP kinase binding.\",\n      \"evidence\": \"Subcellular fractionation, in vitro phosphatase assays, and in vivo substrate selectivity/binding in COS-1 cells\",\n      \"pmids\": [\"9788880\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis of allosteric activation by ERK2 not resolved\",\n        \"Physiological signaling context for ERK2 inactivation not yet addressed\"\n      ]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Refined the substrate specificity question by showing recombinant DUSP7 can also bind phospho-JNK, indicating ERK2 selectivity is not absolute.\",\n      \"evidence\": \"Overexpression with phospho-ERK1/2 Western blot and recombinant pull-down for phospho-JNK in HEK293 cells\",\n      \"pmids\": [\"15081539\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"JNK binding shown by pull-down without functional dephosphorylation in cells\",\n        \"Limited controls for binding specificity\"\n      ]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Defined the gene architecture by identifying a catalytic isoform (PYST2-L) and a catalytically deficient short isoform (PYST2-S), raising the possibility of intramolecular regulation.\",\n      \"evidence\": \"Northern blot, RT-PCR, sequencing, Western blot, and domain analysis\",\n      \"pmids\": [\"14603440\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Negative-regulatory role of PYST2-S inferred but not experimentally demonstrated\",\n        \"Relative abundance and tissue distribution of isoforms unknown\"\n      ]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Linked DUSP7 expression to cell-cycle phase and culture density, hinting at a cell-cycle regulatory function.\",\n      \"evidence\": \"Cell cycle synchronization and expression analysis in K562 cells plus crowding comparisons\",\n      \"pmids\": [\"16386315\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Correlative expression only, no functional rescue or pathway placement\",\n        \"Mechanism connecting crowding to DUSP7 expression unknown\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated a physiological requirement for DUSP7 in meiosis, identifying cPKC isoforms as substrates whose dephosphorylation drives meiotic resumption and proper chromosome alignment.\",\n      \"evidence\": \"RNAi depletion in mouse oocytes with live imaging, Cdk1/CycB activity assays, and cPKC phosphorylation analysis\",\n      \"pmids\": [\"27783954\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct enzyme-substrate dephosphorylation of cPKC not reconstituted in vitro\",\n        \"How cPKC links to the Cdk1/CycB threshold not fully mapped\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Extended DUSP7's regulatory reach to inflammatory metabolism by showing it physically restrains TAK1 to limit NAFLD pathogenesis.\",\n      \"evidence\": \"Co-IP for TAK1 interaction and DUSP7 knockout mice under HFD with pathway and metabolic readouts\",\n      \"pmids\": [\"32311490\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether DUSP7 dephosphorylates TAK1 or acts as a scaffold not distinguished\",\n        \"Single Co-IP without reciprocal biochemical validation of mechanism\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Resolved the mitotic function of ERK2 dephosphorylation, establishing that a MEK/DUSP7 balance setting phospho-ERK2 levels governs chromosome congression.\",\n      \"evidence\": \"Co-IP of DUSP7 with ERK2 in mitosis, catalytic mutant overexpression, siRNA, chemical MEK/ERK inhibition, and chromosome alignment imaging\",\n      \"pmids\": [\"33865857\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"ERK2 substrates relevant to chromosome alignment not identified\",\n        \"Spatial regulation of DUSP7 activity at the spindle/kinetochore unknown\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Positioned DUSP7 as a tumor suppressor in cervical cancer acting through inactivation of the RAS/ERK pathway.\",\n      \"evidence\": \"Overexpression and shRNA knockdown in SIHA cells with HRAS/p-ERK Western blots, in vitro phenotypic assays, and xenografts\",\n      \"pmids\": [\"34435746\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which DUSP7 lowers HRAS not defined\",\n        \"No direct biochemical reconstitution of pathway suppression\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified a FOSL1/DUSP7/PEA15 axis in which transcriptionally induced DUSP7 dephosphorylates PEA15 to promote chemoresistance, adding a new substrate and an upstream regulator.\",\n      \"evidence\": \"qRT-PCR, transcriptional/ChIP-type assays, phospho-PEA15 Western blot, and in vitro/in vivo drug resistance assays in breast cancer cells\",\n      \"pmids\": [\"34907034\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct DUSP7-PEA15 dephosphorylation not reconstituted in vitro\",\n        \"How PEA15 dephosphorylation confers resistance mechanistically unresolved\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DUSP7's substrate choice (ERK2, cPKC, PEA15, TAK1) is partitioned across cell types, cell-cycle phases, and disease contexts, and the structural basis of its allosteric activation, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model of substrate engagement or activation\",\n        \"No unified framework reconciling ERK2-selective and multi-substrate activities\",\n        \"Determinants directing DUSP7 to distinct substrates in different tissues unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 4, 5, 7]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 5, 6]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4, 5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"MAPK1\", \"TAK1\", \"PEA15\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}