{"gene":"SPAG9","run_date":"2026-06-10T07:46:38","timeline":{"discoveries":[{"year":2002,"finding":"JLP (SPAG9) acts as a scaffolding protein that simultaneously tethers JNK and p38MAPK signaling modules with transcription factors Max and c-Myc, as well as their upstream kinases MKK4 and MEKK3, bringing MAPKs and their target transcription factors together for specific signaling pathways.","method":"Co-immunoprecipitation, protein interaction assays, transient transfection","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP establishing multiple binding partners, replicated by multiple subsequent labs","pmids":["12391307"],"is_preprint":false},{"year":2005,"finding":"SPAG9 interacts with JNK isoforms (JNK3 > JNK2 > JNK1) through its JNK-binding domain; both the leucine zipper/coiled-coil domain and transmembrane domain are required for dimerization and proper subcellular localization. No interaction was detected with p38alpha or ERK pathways.","method":"Co-immunoprecipitation, deletion mutant expression, CD spectroscopy, indirect immunofluorescence","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — binding domain mapping with deletion mutants, multiple orthogonal methods in one study","pmids":["15693750"],"is_preprint":false},{"year":2005,"finding":"SPAG9 protein localizes to the acrosomal compartment of intact human spermatozoa and relocates to the equatorial segment after acrosome reaction; anti-SPAG9 antibodies inhibit binding of human spermatozoa to intact human oocytes and hemizona, implicating SPAG9 in sperm-egg interaction.","method":"Indirect immunofluorescence, hemizona assay, zona-free hamster egg penetration test","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with functional antibody inhibition assay, single lab","pmids":["15693750"],"is_preprint":false},{"year":2005,"finding":"JLP (SPAG9) interacts with kinesin light chain 1 (KLC1) through JLP's second leucine zipper domain and a novel leucine zipper-like domain within KLC1's tetratripeptide repeat; mutations at Leu-280, Leu-287, Val-294, and Leu-301 in KLC1 disrupted the interaction. The JLP-KLC1 association is required for formation of a ternary complex with JNK1 and for cytoplasmic localization of JLP.","method":"Yeast two-hybrid screening, co-immunoprecipitation, site-directed mutagenesis, immunofluorescence, dominant-negative KLC1 mislocalization assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — yeast two-hybrid plus reciprocal co-IP plus mutagenesis plus mislocalization rescue, multiple orthogonal methods","pmids":["15987681"],"is_preprint":false},{"year":2006,"finding":"JLP (SPAG9) physically interacts with the intracellular region of cell-surface receptor Cdo, forming a complex with p38alpha/beta MAPK in differentiating myoblasts; Cdo and JLP cooperate to enhance active p38alpha/beta levels. Primary myoblasts from Cdo-/- mice show deficient p38alpha/beta activity that can be rescued by activated MKK6, placing JLP-Cdo at the top of this differentiation-linked p38 signaling cascade.","method":"Co-immunoprecipitation, transfection-based p38 activity assays, Cdo-/- primary myoblasts, activated MKK6 rescue","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP plus genetic rescue in primary KO cells, replicated across multiple approaches","pmids":["17074887"],"is_preprint":false},{"year":2006,"finding":"JLP (SPAG9) physically associates with Galpha13 in RA-stimulated P19 cells and is required for retinoic acid-induced endodermal differentiation; siRNA silencing of JLP abrogates RA-mediated endodermal differentiation analogously to silencing of Galpha13 or JNK.","method":"Co-immunoprecipitation, siRNA knockdown, differentiation assays in P19 cells","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus loss-of-function phenotype, single lab","pmids":["16619266"],"is_preprint":false},{"year":2008,"finding":"Ablation of JLP (Jlp gene disruption) in male mice causes subfertility, with highest JLP expression in elongated spermatids of the testis, indicating a physiological role for JLP in production of functionally normal spermatozoa.","method":"Gene knockout in mice, immunohistochemistry, fertility testing","journal":"Transgenic research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo KO with defined phenotype, single lab","pmids":["18574703"],"is_preprint":false},{"year":2008,"finding":"JLP (SPAG9) interacts with the phosphoinositide kinase PIKfyve through PIKfyve's cpn60_TCP1 consensus sequence and the last 75 residues of the JLP C-terminus; both proteins are required for microtubule-based endosome-to-TGN transport of furin cargo, but not for microtubule-independent Tac-TGN38 trafficking.","method":"Yeast two-hybrid, pulldown, co-immunoprecipitation, siRNA depletion, ectopic rescue, cargo-trafficking assays (Tac-furin, Tac-TGN38)","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal binding assays plus siRNA depletion plus rescue experiments with defined cargo specificity","pmids":["19056739"],"is_preprint":false},{"year":2014,"finding":"JSAP1 and JLP (SPAG9) are required for ARF6 localization to the midbody during cytokinesis; both proteins form a tripartite complex with active ARF6 and kinesin-1 heavy chain (KHC). Mutants of JSAP1 or JLP unable to interact with active ARF6 or KHC failed to rescue ARF6 midbody localization and cytokinesis defects in double-knockout MEFs.","method":"Gene knockout in MEFs, rescue experiments with wild-type and binding-deficient mutants, co-immunoprecipitation, cytokinesis assays","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO plus domain-specific mutant rescue plus co-IP in multiple orthogonal experiments","pmids":["25130574"],"is_preprint":false},{"year":2014,"finding":"JLP (SPAG9) scaffold protein is required for UVB-induced p38 MAPK activation in skin keratinocytes; Jlp knockout mice show impaired p38 activation and resistance to UVB-induced epidermal apoptosis, while topical p38 inhibitor suppresses apoptosis in wild-type but not Jlp KO skin.","method":"Jlp knockout mice (whole-body and keratinocyte-specific), p38 MAPK activity assays, p38 inhibitor treatment, apoptosis assays","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional and whole-body KO with pharmacological rescue confirming p38-dependent pathway placement","pmids":["24520900"],"is_preprint":false},{"year":2015,"finding":"JLP (SPAG9) and JSAP1 are essential regulators of kinesin-1-dependent axonal transport with functional redundancy; double-KO of Jsap1 and Jlp in dorsal telencephalon causes progressive neuron loss. JLP binding to kinesin-1 heavy chain is crucial for kinesin-1–microtubule interactions. Defective transport in dKO neurons leads to JNK hyperactivation via increased intra-axonal Ca2+, and axonal JNK relocalizes to the nucleus via dynein to activate c-Jun and cause neuronal death.","method":"Conditional double-knockout mice, primary neuron culture with inducible gene disruption, gene rescue experiments, kinesin-1 interaction assays","journal":"Cell death and differentiation","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo KO plus primary cell rescue plus mechanistic pathway dissection with multiple orthogonal approaches","pmids":["25571974"],"is_preprint":false},{"year":2015,"finding":"JLP (SPAG9) scaffold protein is required in B lymphocytes for CD40 internalization upon CD154 engagement; JLP deficiency impairs CD40 internalization through a process dependent on Rab5 and dynein-mediated vesicle transport. JLP deficiency also diminishes CD40-dependent MAPK and JNK activation but not NF-κB.","method":"jlp-deficient mice B lymphocytes, dynein inhibitor (ciliobrevin D), co-localization and interaction assays for CD40/JLP/dynein/Rab5, flow cytometry","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic KO in primary cells plus pharmacological inhibitor confirming mechanism, multiple orthogonal readouts","pmids":["25586186"],"is_preprint":false},{"year":2015,"finding":"JSAP1 and JLP (SPAG9) play critical and redundant roles in kinesin-1-dependent axonal transport in cerebellar Purkinje cells; double-KO causes PC axonal dystrophy with selective accumulation of kinesin-1 cargoes in swollen axons and progressive neuronal loss.","method":"Conditional double-knockout mice, immunohistochemistry, axonal transport cargo analysis","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo KO with defined cargo accumulation phenotype, single lab confirming prior finding","pmids":["26320416"],"is_preprint":false},{"year":2016,"finding":"JLP (SPAG9) interacts with JNK in vivo in ovarian cancer cells, and this interaction is stimulated by lysophosphatidic acid (LPA); LPA stimulates translocation of the JLP-JNK complex to the perinuclear region. JLP knockdown abrogates LPA-stimulated JNK activation as well as LPA-stimulated proliferation and invasive migration.","method":"Co-immunoprecipitation, co-localization, shRNA knockdown, xenograft mouse model","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with stimulus-dependent complex formation plus in vivo rescue, single lab","pmids":["27655714"],"is_preprint":false},{"year":2016,"finding":"SPAG9 knockdown in hepatocarcinoma cells suppresses migration and invasion, and inhibits ELK1 expression and p38 signaling; overexpression of ELK1 reverses the inhibitory effects of SPAG9 siRNA, placing SPAG9 upstream of ELK1 in HCC metastatic signaling.","method":"siRNA knockdown, ELK1 overexpression rescue, wound healing/invasion assays, Western blot","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via rescue experiment, single lab","pmids":["27042099"],"is_preprint":false},{"year":2018,"finding":"JLP (SPAG9) protects cancer cells from H2O2-induced cell death through the JLP-JNK signaling pathway; JLP knockdown increases cell death and intracellular ROS levels and attenuates H2O2-induced JNK activation. A JLP mutant lacking the JNK-binding domain failed to rescue cell viability or JNK activation.","method":"siRNA knockdown, rescue with wild-type vs JNK-binding domain mutant JLP, cell viability assays, ROS measurement","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-specific mutant rescue establishes JNK-binding domain requirement, single lab","pmids":["29753743"],"is_preprint":false},{"year":2019,"finding":"JLP (SPAG9) tethers PLK1 to the centrosome upon extracellular stimulation in a p38MAPK- and dynein-dependent manner, leading to centrosome maturation and microtubule array formation; the JLP centrosome localization domain is required for binding the centrosome and forming the JLP focus and microtubule array. This JLP-centrosome function is essential for NF-κB p65 nuclear translocation via microtubule-based transport.","method":"Live imaging, domain deletion analysis, p38 inhibitor treatment, dynein inhibition, NF-κB p65 nuclear transport assays","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal approaches (inhibitors, deletion mutants, live imaging, transport assays) in single rigorous study","pmids":["31803841"],"is_preprint":false},{"year":2019,"finding":"SPAG9 promotes prostate cancer cell proliferation, migration, and cell cycle progression via the MAPK signaling pathway; knockout of SPAG9 suppresses tumor growth in nude mice.","method":"siRNA/overexpression, in vitro proliferation/migration assays, xenograft model, Western blot for MAPK pathway components","journal":"American journal of translational research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — MAPK pathway implication by Western blot only, no specific binding or epistasis experiment, single lab","pmids":["31497238"],"is_preprint":false},{"year":2019,"finding":"SPAG9 interacts with JNK by co-immunoprecipitation in HepG2 liver cancer cells; SPAG9 depletion decreases expression of JNK, p38, and MKK3 but not MKK6, defining a SPAG9/MKK3/p38 signaling axis in liver cancer cell proliferation.","method":"Co-immunoprecipitation, siRNA knockdown, Western blot for pathway components, cell proliferation and apoptosis assays","journal":"Oncology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP confirms direct interaction, pathway specificity (MKK3 vs MKK6) defined by Western blot, single lab","pmids":["30720101"],"is_preprint":false},{"year":2019,"finding":"JLP (SPAG9) regulates lysosome localization (preventing peripheral redistribution) and autophagosome-lysosome fusion; rescue experiments show this requires the JLP-kinesin-1 heavy chain interaction domain but not the dynactin p150Glued or lysosomal transmembrane protein 55b binding regions.","method":"JLP knockdown, lysosome localization imaging, autophagy flux assays, rescue with wild-type and deletion mutants","journal":"Drug discoveries & therapeutics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-specific mutant rescue with functional autophagy readout, single lab","pmids":["32023558"],"is_preprint":false},{"year":2019,"finding":"JLP (SPAG9) mediates protection against curcumin-induced cancer cell death by regulating lysosome positioning and p38 MAPK signaling to induce autophagy; JLP knockdown impairs perinuclear lysosomal accumulation and autophagosome-lysosome fusion. A JLP mutant lacking the p38 MAPK-binding domain failed to rescue cell viability or p38 activation.","method":"JLP knockdown, rescue with wild-type vs p38-binding domain mutant JLP, lysosome localization assays, autophagy assays, cell viability","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-specific mutant rescue identifying p38-binding domain requirement, single lab","pmids":["31787236"],"is_preprint":false},{"year":2022,"finding":"The SPAG9::JAK2 fusion protein (from t(9;17)(p24;q21)) constitutively activates JAK2 kinase domain phosphorylation, phosphorylating STAT3/STAT5/STAT1, and confers IL-3-independent proliferation in Ba/F3 cells. Mutation of JAK2 kinase domain tyrosines abolishes IL-3 independence. STAT1 activates BCL-2 and MCL-1, contributing to aberrant growth; the JAK2-STAT1-BCL-2/MCL-1 axis mediates SPAG9::JAK2-driven proliferation.","method":"Retroviral transduction of Ba/F3 cells, IL-3-independent growth assay, phospho-Western blot, kinase domain mutagenesis, gene expression analysis, JAK inhibitor (ruxolitinib) treatment","journal":"Cancer gene therapy","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — reconstitution in Ba/F3 cells, kinase domain mutagenesis, pharmacological confirmation, multiple pathway readouts","pmids":["35879405"],"is_preprint":false},{"year":2023,"finding":"JLP (SPAG9) tethers Foxk1 transcriptional repressor, preventing Foxk1-mediated repression of N-cadherin; TGF-β1 induces Foxk1 expression, but this is counteracted by JLP. Loss of JLP results in Foxk1 induction and N-cadherin downregulation, promoting partial EMT in renal tubular epithelial cells.","method":"TGF-β1 treatment, JLP knockdown/overexpression, Foxk1 modulation, N-cadherin expression assays, transcription factor binding analysis","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via sequential knockdown/rescue, single lab","pmids":["37013185"],"is_preprint":false},{"year":2023,"finding":"HLA-G functions in an HLA-G/SPAG9/STAT3 axis to promote the establishment of brain metastatic lesions; premetastatic brain metastasis initiating cells exhibit increased HLA-G and SPAG9 expression as part of a shared transcriptomic signature.","method":"RNA sequencing of premetastatic vs macrometastatic brain metastasis initiating cells, functional studies of the HLA-G/SPAG9/STAT3 axis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Low","confidence_rationale":"Tier 3 / Weak — axis identified from transcriptomics with limited mechanistic dissection described in abstract, single lab","pmids":["36780531"],"is_preprint":false},{"year":2026,"finding":"TGF-β1 promotes nuclear translocation of β-catenin and LEF1, which together suppress JLP (SPAG9) transcription, leading to unchecked autophagy and renal fibrosis; inhibition of β-catenin/LEF1 with iCRT3 or LEF1 silencing restores JLP expression and attenuates autophagy and fibrosis in murine models.","method":"CKD patient kidney analysis, murine fibrosis models, HK-2 cell culture, β-catenin/LEF1 inhibitor treatment, LEF1 silencing, JLP expression/autophagy assays","journal":"JCI insight","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo therapeutic rescue plus cell culture mechanistic studies confirming transcriptional regulation, single lab","pmids":["41505216"],"is_preprint":false},{"year":2017,"finding":"JLP (SPAG9) is required for CD4+ T cell TCR-initiated proliferation, IL-2 production, and CD154 induction; JLP deficiency impairs Ca2+ influx and NF-AT activation but not MAPK, NF-κB, or AP-1 signaling pathways upon TCR stimulation.","method":"jlp-deficient mice CD4+ T cells, TCR stimulation, proliferation assay, cytokine measurement, Ca2+ flux assay, NF-AT reporter assay, pathway signaling Western blots","journal":"Molecular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO in primary T cells with specific pathway placement (Ca2+/NF-AT vs MAPK/NF-κB), single lab","pmids":["28521278"],"is_preprint":false}],"current_model":"SPAG9/JLP is a multifunctional scaffolding protein that tethers JNK and p38 MAPK signaling modules (along with upstream kinases MKK4/MEKK3) to transcription factors, interacts with kinesin-1 (via KLC1) and dynein motor complexes to regulate cargo-specific axonal and intracellular transport (including endosome-to-TGN trafficking with PIKfyve, lysosome positioning, and ARF6 delivery to the midbody during cytokinesis), recruits PLK1 to the centrosome for microtubule array formation and NF-κB nuclear import, interfaces with cell-surface receptors (Cdo, CD40) to couple extracellular signals to intracellular MAPK cascades, and also functions in sperm acrosomal localization and sperm-egg interaction."},"narrative":{"mechanistic_narrative":"SPAG9 (JLP) is a multifunctional scaffolding protein that physically tethers stress-activated MAPK signaling modules — JNK and p38 — to upstream kinases (MKK4, MEKK3) and downstream transcription factors (Max, c-Myc), assembling pathway-specific signaling complexes [PMID:12391307]. It binds JNK isoforms (JNK3 > JNK2 > JNK1) through a defined JNK-binding domain, with its leucine-zipper/coiled-coil and transmembrane regions governing dimerization and subcellular localization [PMID:15693750]. A second major role is in microtubule-based transport: SPAG9 binds kinesin light chain 1 (KLC1) and kinesin-1 heavy chain to form motor-cargo complexes, with this interaction required for cytoplasmic localization and for ternary complex assembly with JNK [PMID:15987681]. Through these motor linkages SPAG9, together with the redundant scaffold JSAP1, drives kinesin-1-dependent axonal transport whose loss causes cargo accumulation, JNK hyperactivation, and progressive neuronal death [PMID:25571974, PMID:26320416], delivers active ARF6 to the midbody during cytokinesis [PMID:25130574], controls endosome-to-TGN trafficking of furin cargo via PIKfyve [PMID:19056739], and positions lysosomes to support autophagosome-lysosome fusion [PMID:32023558, PMID:31787236]. SPAG9 couples cell-surface receptors to intracellular cascades, partnering with Cdo to enhance p38α/β activation during myoblast differentiation [PMID:17074887] and mediating CD40 internalization in B lymphocytes through Rab5- and dynein-dependent vesicle transport [PMID:25586186]. Upon extracellular stimulation it recruits PLK1 to the centrosome in a p38- and dynein-dependent manner to drive centrosome maturation, microtubule array formation, and microtubule-based NF-κB p65 nuclear translocation [PMID:31803841]. In immune and developmental contexts SPAG9 also supports TCR-initiated Ca2+ influx and NF-AT activation in CD4+ T cells [PMID:28521278] and retinoic-acid-induced endodermal differentiation downstream of Gα13 [PMID:16619266]. SPAG9 localizes to the sperm acrosome and relocates to the equatorial segment after the acrosome reaction, and its disruption in mice causes male subfertility [PMID:15693750, PMID:18574703]. In disease, SPAG9 sustains MAPK-driven proliferation and invasion across multiple cancers [PMID:27655714, PMID:27042099, PMID:30720101], and a SPAG9::JAK2 fusion from t(9;17) constitutively activates JAK2 to phosphorylate STAT1/3/5 and confer growth-factor-independent proliferation [PMID:35879405]; its transcriptional repression by TGF-β1/β-catenin/LEF1 contributes to renal fibrosis [PMID:41505216].","teleology":[{"year":2002,"claim":"Established SPAG9/JLP as a MAPK scaffold, answering how JNK/p38 modules are spatially organized with upstream kinases and target transcription factors.","evidence":"Co-immunoprecipitation and interaction assays in transfected cells","pmids":["12391307"],"confidence":"High","gaps":["Stoichiometry and structure of the assembled scaffold complex not resolved","Physiological signals triggering complex assembly not defined"]},{"year":2005,"claim":"Mapped the JNK-binding domain and the domains controlling dimerization/localization, defining how SPAG9 selectively engages JNK over p38α or ERK, and identified an acrosomal pool linked to sperm-egg binding.","evidence":"Deletion mutants, CD spectroscopy, immunofluorescence, and hemizona/egg penetration antibody-inhibition assays","pmids":["15693750"],"confidence":"High","gaps":["How a single scaffold reconciles JNK-selectivity with reported p38 binding not resolved","Molecular basis of acrosomal targeting unknown"]},{"year":2005,"claim":"Identified KLC1 as a kinesin-1 link, explaining how SPAG9 achieves cytoplasmic localization and couples motor transport to JNK signaling.","evidence":"Yeast two-hybrid, reciprocal co-IP, site-directed mutagenesis, and dominant-negative mislocalization assays","pmids":["15987681"],"confidence":"High","gaps":["Cargoes transported by the SPAG9-KLC1-JNK complex not defined here","Regulation of motor engagement unknown"]},{"year":2006,"claim":"Connected SPAG9 to cell-surface receptor signaling and developmental differentiation, showing it relays Cdo and Gα13 inputs into p38/JNK cascades.","evidence":"Co-IP, p38 activity assays, Cdo-/- primary myoblasts with MKK6 rescue, and siRNA in P19 cells","pmids":["17074887","16619266"],"confidence":"High","gaps":["Direct vs indirect Cdo-SPAG9 contact regions not fully mapped","Gα13-SPAG9 interaction is single-lab"]},{"year":2008,"claim":"Demonstrated an in vivo physiological requirement, with male subfertility on Jlp disruption establishing relevance to spermatogenesis.","evidence":"Mouse gene knockout, immunohistochemistry, and fertility testing","pmids":["18574703"],"confidence":"Medium","gaps":["Molecular defect in knockout spermatids not defined","Single lab"]},{"year":2008,"claim":"Defined a cargo-specific trafficking function, showing SPAG9-PIKfyve is needed for microtubule-based endosome-to-TGN transport of furin but not microtubule-independent cargo.","evidence":"Yeast two-hybrid, pulldown, co-IP, siRNA depletion with ectopic rescue, and defined cargo-trafficking assays","pmids":["19056739"],"confidence":"High","gaps":["How cargo specificity is encoded not resolved","Link to JNK scaffolding role unclear"]},{"year":2014,"claim":"Extended the motor-scaffold role to cytokinesis and UV stress, showing SPAG9 delivers ARF6 to the midbody and is required for UVB-induced p38-dependent keratinocyte apoptosis.","evidence":"Knockout MEFs/mice with binding-deficient mutant rescue, co-IP, cytokinesis assays, and p38 inhibitor treatment in skin","pmids":["25130574","24520900"],"confidence":"High","gaps":["Functional redundancy with JSAP1 complicates single-protein attribution","Upstream activator of midbody localization not defined"]},{"year":2015,"claim":"Resolved the consequences of transport failure in neurons and the receptor-internalization role, showing kinesin-1-dependent axonal transport requires SPAG9/JSAP1 and that its loss drives dynein-mediated nuclear JNK relocalization and neuronal death, while CD40 internalization needs SPAG9 with Rab5/dynein.","evidence":"Conditional double-knockout mice, primary neuron rescue, kinesin-1 interaction assays, jlp-deficient B cells, and dynein inhibitor (ciliobrevin D)","pmids":["25571974","26320416","25586186"],"confidence":"High","gaps":["Cargo selectivity among kinesin-1 substrates incompletely defined","How the same scaffold engages both kinesin and dynein not mechanistically resolved"]},{"year":2016,"claim":"Linked SPAG9-JNK complex assembly and translocation to oncogenic signaling, showing LPA-stimulated perinuclear relocalization drives cancer proliferation and invasion, with ELK1 placed downstream in HCC.","evidence":"Co-IP, co-localization, shRNA/siRNA knockdown with ELK1 rescue, and xenograft models","pmids":["27655714","27042099"],"confidence":"Medium","gaps":["Single-lab cancer-context findings","Direct vs indirect ELK1 regulation not established"]},{"year":2018,"claim":"Established a cytoprotective JNK-dependent function, showing the JNK-binding domain is required for SPAG9-mediated protection against oxidative cell death.","evidence":"siRNA knockdown with wild-type vs JNK-binding-domain mutant rescue, viability and ROS assays","pmids":["29753743"],"confidence":"Medium","gaps":["Single lab","Mechanism linking JNK scaffolding to ROS control not defined"]},{"year":2019,"claim":"Unified the centrosome, lysosome, and pathway-specificity roles, showing SPAG9 tethers PLK1 to the centrosome for microtubule array formation and NF-κB nuclear import, positions lysosomes via kinesin-1 to enable autophagy, and engages an MKK3/p38 (not MKK6) axis in liver cancer.","evidence":"Live imaging, domain deletion, p38/dynein inhibitors, NF-κB transport assays, autophagy flux with mutant rescue, and co-IP with pathway Western blots","pmids":["31803841","32023558","31787236","30720101","31497238"],"confidence":"High","gaps":["How SPAG9 selects MKK3 over MKK6 not defined","Prostate cancer MAPK link is Western-blot only (Low confidence)"]},{"year":2017,"claim":"Defined a non-MAPK immune role, showing SPAG9 supports TCR-initiated Ca2+ influx and NF-AT activation independently of MAPK/NF-κB/AP-1.","evidence":"jlp-deficient CD4+ T cells, proliferation/cytokine assays, Ca2+ flux, NF-AT reporter, and pathway Western blots","pmids":["28521278"],"confidence":"Medium","gaps":["Mechanism coupling SPAG9 to Ca2+/NF-AT unknown","Single lab"]},{"year":2023,"claim":"Identified a transcription-factor-tethering anti-EMT function, showing SPAG9 sequesters Foxk1 to relieve repression of N-cadherin and counteract TGF-β1-induced partial EMT.","evidence":"TGF-β1 treatment, JLP knockdown/overexpression, Foxk1 modulation, and N-cadherin expression assays","pmids":["37013185"],"confidence":"Medium","gaps":["Direct Foxk1-SPAG9 binding interface not mapped","Single lab"]},{"year":2022,"claim":"Revealed an oncogenic fusion mechanism, showing SPAG9::JAK2 constitutively activates JAK2 to drive STAT1/3/5 phosphorylation and a STAT1-BCL-2/MCL-1 survival axis conferring cytokine-independent growth.","evidence":"Ba/F3 reconstitution, IL-3-independent growth, kinase-domain mutagenesis, and ruxolitinib treatment","pmids":["35879405"],"confidence":"High","gaps":["Contribution of SPAG9 dimerization domains to fusion activation not dissected","Prevalence in patients not addressed by these experiments"]},{"year":2026,"claim":"Placed SPAG9 as a transcriptionally regulated brake on fibrosis-associated autophagy, showing TGF-β1/β-catenin/LEF1 represses SPAG9 to unleash autophagy and renal fibrosis, reversible by β-catenin/LEF1 inhibition.","evidence":"CKD patient tissue, murine fibrosis models, HK-2 cells, iCRT3/LEF1 silencing, and JLP expression/autophagy assays","pmids":["41505216"],"confidence":"Medium","gaps":["Direct LEF1 binding at the SPAG9 promoter not fully characterized","Single lab"]},{"year":null,"claim":"How a single scaffold integrates seemingly contradictory roles — selective JNK vs p38 binding, simultaneous engagement of kinesin and dynein, and pro- vs anti-survival outputs — across cell types remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of the full SPAG9 scaffold or its cargo/motor interfaces","Context-determinants selecting among partners not defined","Relative contributions of SPAG9 vs redundant JSAP1 in vivo incompletely partitioned"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,3,8]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[3,8,10,19]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,4,16]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,3]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[16]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[19,20]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[7,11]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[10,16]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,4,16]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[7,8,11,19]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[19,20,24]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[11,25]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,5]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[8]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[21]}],"complexes":[],"partners":["JNK3","MKK4","MEKK3","KLC1","PIKFYVE","ARF6","PLK1","CDO"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60271","full_name":"C-Jun-amino-terminal kinase-interacting protein 4","aliases":["Cancer/testis antigen 89","CT89","Human lung cancer oncogene 6 protein","HLC-6","JNK-associated leucine-zipper protein","JLP","Mitogen-activated protein kinase 8-interacting protein 4","Proliferation-inducing protein 6","Protein highly expressed in testis","PHET","Sperm surface protein","Sperm-associated antigen 9","Sperm-specific protein","Sunday driver 1"],"length_aa":1321,"mass_kda":146.2,"function":"The JNK-interacting protein (JIP) group of scaffold proteins selectively mediates JNK signaling by aggregating specific components of the MAPK cascade to form a functional JNK signaling module (PubMed:14743216). Regulates lysosomal positioning by acting as an adapter protein which links PIP4P1-positive lysosomes to the dynein-dynactin complex (PubMed:29146937). Assists PIKFYVE selective functionality in microtubule-based endosome-to-TGN trafficking (By similarity)","subcellular_location":"Cytoplasmic vesicle, secretory vesicle, acrosome","url":"https://www.uniprot.org/uniprotkb/O60271/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/SPAG9","classification":"Not Classified","n_dependent_lines":71,"n_total_lines":1208,"dependency_fraction":0.058774834437086095},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"EMC9","stoichiometry":0.2},{"gene":"NCAPH","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/SPAG9","total_profiled":1310},"omim":[{"mim_id":"605430","title":"SPERM-ASSOCIATED ANTIGEN 9; SPAG9","url":"https://www.omim.org/entry/605430"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Centriolar satellite","reliability":"Enhanced"},{"location":"Cytosol","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/SPAG9"},"hgnc":{"alias_symbol":["HSS","SYD1","KIAA0516","MGC14967","MGC74461","MGC117291","JLP","PHET","HLC4","FLJ13450","FLJ14006","FLJ26141","FLJ34602","CT89","JIP4","JIP-4","PIG6"],"prev_symbol":[]},"alphafold":{"accession":"O60271","domains":[{"cath_id":"-","chopping":"642-678_1131-1175_1199-1238_1266-1292_1303-1321","consensus_level":"medium","plddt":78.3202,"start":642,"end":1321},{"cath_id":"1.20.5","chopping":"23-166","consensus_level":"high","plddt":91.6635,"start":23,"end":166}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60271","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60271-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60271-F1-predicted_aligned_error_v6.png","plddt_mean":64.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=SPAG9","jax_strain_url":"https://www.jax.org/strain/search?query=SPAG9"},"sequence":{"accession":"O60271","fasta_url":"https://rest.uniprot.org/uniprotkb/O60271.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60271/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60271"}},"corpus_meta":[{"pmid":"12391307","id":"PMC_12391307","title":"JLP: A scaffolding protein that tethers JNK/p38MAPK signaling modules and transcription factors.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12391307","citation_count":118,"is_preprint":false},{"pmid":"17074887","id":"PMC_17074887","title":"Activation of p38alpha/beta MAPK in myogenesis via binding of the scaffold protein JLP to the cell surface protein Cdo.","date":"2006","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/17074887","citation_count":103,"is_preprint":false},{"pmid":"15693750","id":"PMC_15693750","title":"Characterization of a novel human sperm-associated antigen 9 (SPAG9) having structural homology with c-Jun N-terminal kinase-interacting protein.","date":"2005","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/15693750","citation_count":74,"is_preprint":false},{"pmid":"30755596","id":"PMC_30755596","title":"LncRNA H19 promotes the committed differentiation of stem cells from apical papilla via miR-141/SPAG9 pathway.","date":"2019","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/30755596","citation_count":53,"is_preprint":false},{"pmid":"15987681","id":"PMC_15987681","title":"JLP associates with kinesin light chain 1 through a novel leucine zipper-like domain.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15987681","citation_count":51,"is_preprint":false},{"pmid":"26797273","id":"PMC_26797273","title":"MicroRNA-200a-3p suppresses tumor proliferation and induces apoptosis by targeting SPAG9 in renal cell carcinoma.","date":"2016","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/26797273","citation_count":44,"is_preprint":false},{"pmid":"25571974","id":"PMC_25571974","title":"JSAP1/JIP3 and JLP regulate kinesin-1-dependent axonal transport to prevent neuronal degeneration.","date":"2015","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/25571974","citation_count":37,"is_preprint":false},{"pmid":"24330581","id":"PMC_24330581","title":"Down regulation of SPAG9 reduces growth and invasive potential of triple-negative breast cancer cells: possible implications in targeted therapy.","date":"2013","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/24330581","citation_count":35,"is_preprint":false},{"pmid":"31485599","id":"PMC_31485599","title":"LncRNA NEAT1 enhances the resistance of anaplastic thyroid carcinoma cells to cisplatin by sponging miR‑9‑5p and regulating SPAG9 expression.","date":"2019","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/31485599","citation_count":34,"is_preprint":false},{"pmid":"23711689","id":"PMC_23711689","title":"Clinical significance and biological roles of SPAG9 overexpression in non-small cell lung cancer.","date":"2013","source":"Lung cancer (Amsterdam, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/23711689","citation_count":32,"is_preprint":false},{"pmid":"18574703","id":"PMC_18574703","title":"Ablation of the scaffold protein JLP causes reduced fertility in male mice.","date":"2008","source":"Transgenic research","url":"https://pubmed.ncbi.nlm.nih.gov/18574703","citation_count":31,"is_preprint":false},{"pmid":"19056739","id":"PMC_19056739","title":"Kinesin adapter JLP links PIKfyve to microtubule-based endosome-to-trans-Golgi network traffic of furin.","date":"2008","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19056739","citation_count":31,"is_preprint":false},{"pmid":"19813278","id":"PMC_19813278","title":"Small interfering RNA-mediated down-regulation of SPAG9 inhibits cervical tumor growth.","date":"2009","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19813278","citation_count":30,"is_preprint":false},{"pmid":"23696027","id":"PMC_23696027","title":"SPAG9 is overexpressed in human astrocytoma and promotes cell proliferation and invasion.","date":"2013","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/23696027","citation_count":30,"is_preprint":false},{"pmid":"16077255","id":"PMC_16077255","title":"Sperm associated antigen 9 (SPAG9): a new member of c-Jun NH2 -terminal kinase (JNK) interacting protein exclusively expressed in testis.","date":"2005","source":"The Keio journal of medicine","url":"https://pubmed.ncbi.nlm.nih.gov/16077255","citation_count":23,"is_preprint":false},{"pmid":"25586186","id":"PMC_25586186","title":"Scaffold protein JLP is critical for CD40 signaling in B lymphocytes.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25586186","citation_count":20,"is_preprint":false},{"pmid":"28028759","id":"PMC_28028759","title":"The expression of DAMP proteins HSP70 and cancer-testis antigen SPAG9 in peripheral blood of patients with HCC and lung cancer.","date":"2016","source":"Cell stress & chaperones","url":"https://pubmed.ncbi.nlm.nih.gov/28028759","citation_count":20,"is_preprint":false},{"pmid":"16619266","id":"PMC_16619266","title":"Endodermal differentiation of murine embryonic carcinoma cells by retinoic acid requires JLP, a JNK-scaffolding protein.","date":"2006","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16619266","citation_count":20,"is_preprint":false},{"pmid":"16061308","id":"PMC_16061308","title":"Characterization of immune response in mice to plasmid DNA encoding human sperm associated antigen 9 (SPAG9).","date":"2005","source":"Vaccine","url":"https://pubmed.ncbi.nlm.nih.gov/16061308","citation_count":20,"is_preprint":false},{"pmid":"24740566","id":"PMC_24740566","title":"SPAG9 is overexpressed in human prostate cancer and promotes cancer cell proliferation.","date":"2014","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24740566","citation_count":18,"is_preprint":false},{"pmid":"27042099","id":"PMC_27042099","title":"SPAG9 is involved in hepatocarcinoma cell migration and invasion via modulation of ELK1 expression.","date":"2016","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/27042099","citation_count":18,"is_preprint":false},{"pmid":"15293220","id":"PMC_15293220","title":"Isolation and characterization of a haploid germ cell specific sperm associated antigen 9 (SPAG9) from the baboon.","date":"2004","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15293220","citation_count":18,"is_preprint":false},{"pmid":"16143403","id":"PMC_16143403","title":"Immunogenicity and contraceptive potential of recombinant human sperm associated antigen (SPAG9).","date":"2005","source":"Journal of reproductive immunology","url":"https://pubmed.ncbi.nlm.nih.gov/16143403","citation_count":17,"is_preprint":false},{"pmid":"27449044","id":"PMC_27449044","title":"Sperm-associated antigen 9 (SPAG9) promotes the survival and tumor growth of triple-negative breast cancer cells.","date":"2016","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27449044","citation_count":16,"is_preprint":false},{"pmid":"26934841","id":"PMC_26934841","title":"Cancer testis antigen SPAG9 is a promising marker for the diagnosis and treatment of lung cancer.","date":"2016","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/26934841","citation_count":15,"is_preprint":false},{"pmid":"25951811","id":"PMC_25951811","title":"Identification of SPAG9 as a novel JAK2 fusion partner gene in pediatric acute lymphoblastic leukemia with t(9;17)(p24;q21).","date":"2015","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/25951811","citation_count":15,"is_preprint":false},{"pmid":"31497238","id":"PMC_31497238","title":"SPAG9 promotes prostate cancer proliferation and metastasis via MAPK signaling pathway.","date":"2019","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/31497238","citation_count":14,"is_preprint":false},{"pmid":"25310386","id":"PMC_25310386","title":"SPAG9 expression is increased in human prostate cancer and promotes cell motility, invasion and angiogenesis in vitro.","date":"2014","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/25310386","citation_count":14,"is_preprint":false},{"pmid":"27655714","id":"PMC_27655714","title":"Aberrant expression of JNK-associated leucine-zipper protein, JLP, promotes accelerated growth of ovarian cancer.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27655714","citation_count":12,"is_preprint":false},{"pmid":"29753743","id":"PMC_29753743","title":"JLP-JNK signaling protects cancer cells from reactive oxygen species-induced cell death.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29753743","citation_count":12,"is_preprint":false},{"pmid":"29949157","id":"PMC_29949157","title":"MiR-524 inhibits cell proliferation and induces cell apoptosis in thyroid cancer via targeting SPAG9.","date":"2018","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29949157","citation_count":12,"is_preprint":false},{"pmid":"36780531","id":"PMC_36780531","title":"An HLA-G/SPAG9/STAT3 axis promotes brain metastases.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/36780531","citation_count":11,"is_preprint":false},{"pmid":"30662804","id":"PMC_30662804","title":"SPAG9 regulates HEF1 expression and drives EMT in bladder transitional cell carcinoma via rac1 signaling pathway.","date":"2018","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/30662804","citation_count":11,"is_preprint":false},{"pmid":"29745297","id":"PMC_29745297","title":"Sperm associated antigen 9 (SPAG9) a promising therapeutic target of ovarian carcinoma.","date":"2018","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29745297","citation_count":11,"is_preprint":false},{"pmid":"25130574","id":"PMC_25130574","title":"JSAP1 and JLP are required for ARF6 localization to the midbody in cytokinesis.","date":"2014","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/25130574","citation_count":11,"is_preprint":false},{"pmid":"34097562","id":"PMC_34097562","title":"ZSCAN16-AS1 expedites hepatocellular carcinoma progression via modulating the miR-181c-5p/SPAG9 axis to activate the JNK pathway.","date":"2021","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/34097562","citation_count":11,"is_preprint":false},{"pmid":"37357722","id":"PMC_37357722","title":"The bioceramic sealer iRoot SP promotes osteogenic differentiation of human stem cells from apical papilla via miR-141-3p/SPAG9/MAPK signalling pathway.","date":"2023","source":"International endodontic journal","url":"https://pubmed.ncbi.nlm.nih.gov/37357722","citation_count":11,"is_preprint":false},{"pmid":"30720101","id":"PMC_30720101","title":"SPAG9/MKK3/p38 axis is a novel therapeutic target for liver cancer.","date":"2019","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/30720101","citation_count":10,"is_preprint":false},{"pmid":"32023558","id":"PMC_32023558","title":"Functional role of c-Jun NH2-terminal kinase-associated leucine zipper protein (JLP) in lysosome localization and autophagy.","date":"2020","source":"Drug discoveries & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/32023558","citation_count":9,"is_preprint":false},{"pmid":"23182713","id":"PMC_23182713","title":"A novel role of the scaffolding protein JLP in tuning CD40-induced activation of dendritic cells.","date":"2012","source":"Immunobiology","url":"https://pubmed.ncbi.nlm.nih.gov/23182713","citation_count":9,"is_preprint":false},{"pmid":"15736136","id":"PMC_15736136","title":"Molecular cloning and characterization of the macaque sperm associated antigen 9 (SPAG9): an orthologue of human SPAG9 gene.","date":"2005","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/15736136","citation_count":9,"is_preprint":false},{"pmid":"25941602","id":"PMC_25941602","title":"Sperm associated antigen 9 (SPAG9) expression and humoral response in benign and malignant salivary gland tumors.","date":"2014","source":"Oncoimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/25941602","citation_count":8,"is_preprint":false},{"pmid":"31803841","id":"PMC_31803841","title":"JLP-centrosome is essential for the microtubule-mediated nucleocytoplasmic transport induced by extracellular stimuli.","date":"2019","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/31803841","citation_count":7,"is_preprint":false},{"pmid":"31787236","id":"PMC_31787236","title":"Protective role of c-Jun NH2-terminal kinase-associated leucine zipper protein (JLP) in curcumin-induced cancer cell death.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31787236","citation_count":7,"is_preprint":false},{"pmid":"27698841","id":"PMC_27698841","title":"SPAG9 is overexpressed in osteosarcoma, and regulates cell proliferation and invasion through regulation of JunD.","date":"2016","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/27698841","citation_count":6,"is_preprint":false},{"pmid":"26893659","id":"PMC_26893659","title":"SPAG9 controls the cell motility, invasion and angiogenesis of human osteosarcoma cells.","date":"2015","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26893659","citation_count":6,"is_preprint":false},{"pmid":"37013185","id":"PMC_37013185","title":"JLP/Foxk1/N-cadherin axis fosters a partial epithelial-mesenchymal transition state in epithelial tubular cells.","date":"2023","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/37013185","citation_count":6,"is_preprint":false},{"pmid":"35879405","id":"PMC_35879405","title":"The combination of ruxolitinib and Bcl-2/Mcl-1 inhibitors has a synergistic effect on leukemic cells carrying a SPAG9::JAK2 fusion.","date":"2022","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/35879405","citation_count":6,"is_preprint":false},{"pmid":"24520900","id":"PMC_24520900","title":"The scaffold protein JLP plays a key role in regulating ultraviolet B-induced apoptosis in mice.","date":"2014","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/24520900","citation_count":5,"is_preprint":false},{"pmid":"28521278","id":"PMC_28521278","title":"Scaffold protein JLP mediates TCR-initiated CD4+T cell activation and CD154 expression.","date":"2017","source":"Molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/28521278","citation_count":4,"is_preprint":false},{"pmid":"26320416","id":"PMC_26320416","title":"Critical role of JSAP1 and JLP in axonal transport in the cerebellar Purkinje cells of mice.","date":"2015","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/26320416","citation_count":4,"is_preprint":false},{"pmid":"27352556","id":"PMC_27352556","title":"SPAG9 promotes endometrial carcinoma cell invasion through regulation of genes related to the epithelial-mesenchymal transition.","date":"2016","source":"European journal of gynaecological oncology","url":"https://pubmed.ncbi.nlm.nih.gov/27352556","citation_count":3,"is_preprint":false},{"pmid":"33716240","id":"PMC_33716240","title":"c-Jun NH2-terminal kinase (JNK)/stress-activated protein kinase-associated protein 1 (JSAP1) attenuates curcumin-induced cell death differently from its family member, JNK-associated leucine zipper protein (JLP).","date":"2021","source":"Drug discoveries & therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/33716240","citation_count":3,"is_preprint":false},{"pmid":"32503577","id":"PMC_32503577","title":"miR-874 inhibits gastric cancer cell proliferation by targeting SPAG9.","date":"2020","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/32503577","citation_count":3,"is_preprint":false},{"pmid":"39846792","id":"PMC_39846792","title":"Truncated SPAG9 as a novel candidate gene for a new syndrome: Coarse facial features, albinism, cataract and developmental delay (CACD syndrome).","date":"2025","source":"Genetics and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/39846792","citation_count":3,"is_preprint":false},{"pmid":"34561997","id":"PMC_34561997","title":"Oncolytic adenovirus carrying SPAG9-shRNA enhanced the efficacy of docetaxel for advanced prostate cancer.","date":"2022","source":"Anti-cancer drugs","url":"https://pubmed.ncbi.nlm.nih.gov/34561997","citation_count":2,"is_preprint":false},{"pmid":"35535313","id":"PMC_35535313","title":"Oncolytic Adenovirus with SPAG9 shRNA Driven by DD3 Promoter Improved the Efficacy of Docetaxil for Prostate Cancer.","date":"2022","source":"Journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/35535313","citation_count":2,"is_preprint":false},{"pmid":"24783006","id":"PMC_24783006","title":"Assessment of SPAG9 Transcript in Fine Needle Aspirates of Thyroid Nodules.","date":"2012","source":"European thyroid journal","url":"https://pubmed.ncbi.nlm.nih.gov/24783006","citation_count":2,"is_preprint":false},{"pmid":"34380828","id":"PMC_34380828","title":"Significance of cancer testis-associated antigens (SPAG9 and FBXO39) in colon cancer.","date":"2022","source":"Indian journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/34380828","citation_count":1,"is_preprint":false},{"pmid":"19947555","id":"PMC_19947555","title":"[Expression and location of SPAG9 in human ejaculated spermatozoa].","date":"2009","source":"Zhonghua nan ke xue = National journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/19947555","citation_count":1,"is_preprint":false},{"pmid":"41505216","id":"PMC_41505216","title":"A TGF-β1/LEF1/β-catenin/JLP network motif regulates autophagy and tubule injury in renal fibrosis.","date":"2026","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/41505216","citation_count":0,"is_preprint":false},{"pmid":"35383860","id":"PMC_35383860","title":"[Corrigendum] SPAG9 expression is increased in human prostate cancer and promotes cell motility, invasion and angiogenesis in vitro.","date":"2022","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/35383860","citation_count":0,"is_preprint":false},{"pmid":"34919215","id":"PMC_34919215","title":"MiR-524 inhibits cell proliferation and induces cell apoptosis in thyroid cancer via targeting SPAG9.","date":"2021","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34919215","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":31959,"output_tokens":6346,"usd":0.095533,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15164,"output_tokens":5405,"usd":0.105472,"stage2_stop_reason":"end_turn"},"total_usd":0.201005,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"JLP (SPAG9) acts as a scaffolding protein that simultaneously tethers JNK and p38MAPK signaling modules with transcription factors Max and c-Myc, as well as their upstream kinases MKK4 and MEKK3, bringing MAPKs and their target transcription factors together for specific signaling pathways.\",\n      \"method\": \"Co-immunoprecipitation, protein interaction assays, transient transfection\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP establishing multiple binding partners, replicated by multiple subsequent labs\",\n      \"pmids\": [\"12391307\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"SPAG9 interacts with JNK isoforms (JNK3 > JNK2 > JNK1) through its JNK-binding domain; both the leucine zipper/coiled-coil domain and transmembrane domain are required for dimerization and proper subcellular localization. No interaction was detected with p38alpha or ERK pathways.\",\n      \"method\": \"Co-immunoprecipitation, deletion mutant expression, CD spectroscopy, indirect immunofluorescence\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — binding domain mapping with deletion mutants, multiple orthogonal methods in one study\",\n      \"pmids\": [\"15693750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"SPAG9 protein localizes to the acrosomal compartment of intact human spermatozoa and relocates to the equatorial segment after acrosome reaction; anti-SPAG9 antibodies inhibit binding of human spermatozoa to intact human oocytes and hemizona, implicating SPAG9 in sperm-egg interaction.\",\n      \"method\": \"Indirect immunofluorescence, hemizona assay, zona-free hamster egg penetration test\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with functional antibody inhibition assay, single lab\",\n      \"pmids\": [\"15693750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"JLP (SPAG9) interacts with kinesin light chain 1 (KLC1) through JLP's second leucine zipper domain and a novel leucine zipper-like domain within KLC1's tetratripeptide repeat; mutations at Leu-280, Leu-287, Val-294, and Leu-301 in KLC1 disrupted the interaction. The JLP-KLC1 association is required for formation of a ternary complex with JNK1 and for cytoplasmic localization of JLP.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation, site-directed mutagenesis, immunofluorescence, dominant-negative KLC1 mislocalization assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — yeast two-hybrid plus reciprocal co-IP plus mutagenesis plus mislocalization rescue, multiple orthogonal methods\",\n      \"pmids\": [\"15987681\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"JLP (SPAG9) physically interacts with the intracellular region of cell-surface receptor Cdo, forming a complex with p38alpha/beta MAPK in differentiating myoblasts; Cdo and JLP cooperate to enhance active p38alpha/beta levels. Primary myoblasts from Cdo-/- mice show deficient p38alpha/beta activity that can be rescued by activated MKK6, placing JLP-Cdo at the top of this differentiation-linked p38 signaling cascade.\",\n      \"method\": \"Co-immunoprecipitation, transfection-based p38 activity assays, Cdo-/- primary myoblasts, activated MKK6 rescue\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP plus genetic rescue in primary KO cells, replicated across multiple approaches\",\n      \"pmids\": [\"17074887\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"JLP (SPAG9) physically associates with Galpha13 in RA-stimulated P19 cells and is required for retinoic acid-induced endodermal differentiation; siRNA silencing of JLP abrogates RA-mediated endodermal differentiation analogously to silencing of Galpha13 or JNK.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, differentiation assays in P19 cells\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus loss-of-function phenotype, single lab\",\n      \"pmids\": [\"16619266\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Ablation of JLP (Jlp gene disruption) in male mice causes subfertility, with highest JLP expression in elongated spermatids of the testis, indicating a physiological role for JLP in production of functionally normal spermatozoa.\",\n      \"method\": \"Gene knockout in mice, immunohistochemistry, fertility testing\",\n      \"journal\": \"Transgenic research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo KO with defined phenotype, single lab\",\n      \"pmids\": [\"18574703\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"JLP (SPAG9) interacts with the phosphoinositide kinase PIKfyve through PIKfyve's cpn60_TCP1 consensus sequence and the last 75 residues of the JLP C-terminus; both proteins are required for microtubule-based endosome-to-TGN transport of furin cargo, but not for microtubule-independent Tac-TGN38 trafficking.\",\n      \"method\": \"Yeast two-hybrid, pulldown, co-immunoprecipitation, siRNA depletion, ectopic rescue, cargo-trafficking assays (Tac-furin, Tac-TGN38)\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal binding assays plus siRNA depletion plus rescue experiments with defined cargo specificity\",\n      \"pmids\": [\"19056739\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"JSAP1 and JLP (SPAG9) are required for ARF6 localization to the midbody during cytokinesis; both proteins form a tripartite complex with active ARF6 and kinesin-1 heavy chain (KHC). Mutants of JSAP1 or JLP unable to interact with active ARF6 or KHC failed to rescue ARF6 midbody localization and cytokinesis defects in double-knockout MEFs.\",\n      \"method\": \"Gene knockout in MEFs, rescue experiments with wild-type and binding-deficient mutants, co-immunoprecipitation, cytokinesis assays\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO plus domain-specific mutant rescue plus co-IP in multiple orthogonal experiments\",\n      \"pmids\": [\"25130574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"JLP (SPAG9) scaffold protein is required for UVB-induced p38 MAPK activation in skin keratinocytes; Jlp knockout mice show impaired p38 activation and resistance to UVB-induced epidermal apoptosis, while topical p38 inhibitor suppresses apoptosis in wild-type but not Jlp KO skin.\",\n      \"method\": \"Jlp knockout mice (whole-body and keratinocyte-specific), p38 MAPK activity assays, p38 inhibitor treatment, apoptosis assays\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional and whole-body KO with pharmacological rescue confirming p38-dependent pathway placement\",\n      \"pmids\": [\"24520900\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"JLP (SPAG9) and JSAP1 are essential regulators of kinesin-1-dependent axonal transport with functional redundancy; double-KO of Jsap1 and Jlp in dorsal telencephalon causes progressive neuron loss. JLP binding to kinesin-1 heavy chain is crucial for kinesin-1–microtubule interactions. Defective transport in dKO neurons leads to JNK hyperactivation via increased intra-axonal Ca2+, and axonal JNK relocalizes to the nucleus via dynein to activate c-Jun and cause neuronal death.\",\n      \"method\": \"Conditional double-knockout mice, primary neuron culture with inducible gene disruption, gene rescue experiments, kinesin-1 interaction assays\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo KO plus primary cell rescue plus mechanistic pathway dissection with multiple orthogonal approaches\",\n      \"pmids\": [\"25571974\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"JLP (SPAG9) scaffold protein is required in B lymphocytes for CD40 internalization upon CD154 engagement; JLP deficiency impairs CD40 internalization through a process dependent on Rab5 and dynein-mediated vesicle transport. JLP deficiency also diminishes CD40-dependent MAPK and JNK activation but not NF-κB.\",\n      \"method\": \"jlp-deficient mice B lymphocytes, dynein inhibitor (ciliobrevin D), co-localization and interaction assays for CD40/JLP/dynein/Rab5, flow cytometry\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic KO in primary cells plus pharmacological inhibitor confirming mechanism, multiple orthogonal readouts\",\n      \"pmids\": [\"25586186\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"JSAP1 and JLP (SPAG9) play critical and redundant roles in kinesin-1-dependent axonal transport in cerebellar Purkinje cells; double-KO causes PC axonal dystrophy with selective accumulation of kinesin-1 cargoes in swollen axons and progressive neuronal loss.\",\n      \"method\": \"Conditional double-knockout mice, immunohistochemistry, axonal transport cargo analysis\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo KO with defined cargo accumulation phenotype, single lab confirming prior finding\",\n      \"pmids\": [\"26320416\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"JLP (SPAG9) interacts with JNK in vivo in ovarian cancer cells, and this interaction is stimulated by lysophosphatidic acid (LPA); LPA stimulates translocation of the JLP-JNK complex to the perinuclear region. JLP knockdown abrogates LPA-stimulated JNK activation as well as LPA-stimulated proliferation and invasive migration.\",\n      \"method\": \"Co-immunoprecipitation, co-localization, shRNA knockdown, xenograft mouse model\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with stimulus-dependent complex formation plus in vivo rescue, single lab\",\n      \"pmids\": [\"27655714\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"SPAG9 knockdown in hepatocarcinoma cells suppresses migration and invasion, and inhibits ELK1 expression and p38 signaling; overexpression of ELK1 reverses the inhibitory effects of SPAG9 siRNA, placing SPAG9 upstream of ELK1 in HCC metastatic signaling.\",\n      \"method\": \"siRNA knockdown, ELK1 overexpression rescue, wound healing/invasion assays, Western blot\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via rescue experiment, single lab\",\n      \"pmids\": [\"27042099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"JLP (SPAG9) protects cancer cells from H2O2-induced cell death through the JLP-JNK signaling pathway; JLP knockdown increases cell death and intracellular ROS levels and attenuates H2O2-induced JNK activation. A JLP mutant lacking the JNK-binding domain failed to rescue cell viability or JNK activation.\",\n      \"method\": \"siRNA knockdown, rescue with wild-type vs JNK-binding domain mutant JLP, cell viability assays, ROS measurement\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-specific mutant rescue establishes JNK-binding domain requirement, single lab\",\n      \"pmids\": [\"29753743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"JLP (SPAG9) tethers PLK1 to the centrosome upon extracellular stimulation in a p38MAPK- and dynein-dependent manner, leading to centrosome maturation and microtubule array formation; the JLP centrosome localization domain is required for binding the centrosome and forming the JLP focus and microtubule array. This JLP-centrosome function is essential for NF-κB p65 nuclear translocation via microtubule-based transport.\",\n      \"method\": \"Live imaging, domain deletion analysis, p38 inhibitor treatment, dynein inhibition, NF-κB p65 nuclear transport assays\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal approaches (inhibitors, deletion mutants, live imaging, transport assays) in single rigorous study\",\n      \"pmids\": [\"31803841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SPAG9 promotes prostate cancer cell proliferation, migration, and cell cycle progression via the MAPK signaling pathway; knockout of SPAG9 suppresses tumor growth in nude mice.\",\n      \"method\": \"siRNA/overexpression, in vitro proliferation/migration assays, xenograft model, Western blot for MAPK pathway components\",\n      \"journal\": \"American journal of translational research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — MAPK pathway implication by Western blot only, no specific binding or epistasis experiment, single lab\",\n      \"pmids\": [\"31497238\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SPAG9 interacts with JNK by co-immunoprecipitation in HepG2 liver cancer cells; SPAG9 depletion decreases expression of JNK, p38, and MKK3 but not MKK6, defining a SPAG9/MKK3/p38 signaling axis in liver cancer cell proliferation.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, Western blot for pathway components, cell proliferation and apoptosis assays\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP confirms direct interaction, pathway specificity (MKK3 vs MKK6) defined by Western blot, single lab\",\n      \"pmids\": [\"30720101\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"JLP (SPAG9) regulates lysosome localization (preventing peripheral redistribution) and autophagosome-lysosome fusion; rescue experiments show this requires the JLP-kinesin-1 heavy chain interaction domain but not the dynactin p150Glued or lysosomal transmembrane protein 55b binding regions.\",\n      \"method\": \"JLP knockdown, lysosome localization imaging, autophagy flux assays, rescue with wild-type and deletion mutants\",\n      \"journal\": \"Drug discoveries & therapeutics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-specific mutant rescue with functional autophagy readout, single lab\",\n      \"pmids\": [\"32023558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"JLP (SPAG9) mediates protection against curcumin-induced cancer cell death by regulating lysosome positioning and p38 MAPK signaling to induce autophagy; JLP knockdown impairs perinuclear lysosomal accumulation and autophagosome-lysosome fusion. A JLP mutant lacking the p38 MAPK-binding domain failed to rescue cell viability or p38 activation.\",\n      \"method\": \"JLP knockdown, rescue with wild-type vs p38-binding domain mutant JLP, lysosome localization assays, autophagy assays, cell viability\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-specific mutant rescue identifying p38-binding domain requirement, single lab\",\n      \"pmids\": [\"31787236\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The SPAG9::JAK2 fusion protein (from t(9;17)(p24;q21)) constitutively activates JAK2 kinase domain phosphorylation, phosphorylating STAT3/STAT5/STAT1, and confers IL-3-independent proliferation in Ba/F3 cells. Mutation of JAK2 kinase domain tyrosines abolishes IL-3 independence. STAT1 activates BCL-2 and MCL-1, contributing to aberrant growth; the JAK2-STAT1-BCL-2/MCL-1 axis mediates SPAG9::JAK2-driven proliferation.\",\n      \"method\": \"Retroviral transduction of Ba/F3 cells, IL-3-independent growth assay, phospho-Western blot, kinase domain mutagenesis, gene expression analysis, JAK inhibitor (ruxolitinib) treatment\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — reconstitution in Ba/F3 cells, kinase domain mutagenesis, pharmacological confirmation, multiple pathway readouts\",\n      \"pmids\": [\"35879405\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"JLP (SPAG9) tethers Foxk1 transcriptional repressor, preventing Foxk1-mediated repression of N-cadherin; TGF-β1 induces Foxk1 expression, but this is counteracted by JLP. Loss of JLP results in Foxk1 induction and N-cadherin downregulation, promoting partial EMT in renal tubular epithelial cells.\",\n      \"method\": \"TGF-β1 treatment, JLP knockdown/overexpression, Foxk1 modulation, N-cadherin expression assays, transcription factor binding analysis\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via sequential knockdown/rescue, single lab\",\n      \"pmids\": [\"37013185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HLA-G functions in an HLA-G/SPAG9/STAT3 axis to promote the establishment of brain metastatic lesions; premetastatic brain metastasis initiating cells exhibit increased HLA-G and SPAG9 expression as part of a shared transcriptomic signature.\",\n      \"method\": \"RNA sequencing of premetastatic vs macrometastatic brain metastasis initiating cells, functional studies of the HLA-G/SPAG9/STAT3 axis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — axis identified from transcriptomics with limited mechanistic dissection described in abstract, single lab\",\n      \"pmids\": [\"36780531\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TGF-β1 promotes nuclear translocation of β-catenin and LEF1, which together suppress JLP (SPAG9) transcription, leading to unchecked autophagy and renal fibrosis; inhibition of β-catenin/LEF1 with iCRT3 or LEF1 silencing restores JLP expression and attenuates autophagy and fibrosis in murine models.\",\n      \"method\": \"CKD patient kidney analysis, murine fibrosis models, HK-2 cell culture, β-catenin/LEF1 inhibitor treatment, LEF1 silencing, JLP expression/autophagy assays\",\n      \"journal\": \"JCI insight\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo therapeutic rescue plus cell culture mechanistic studies confirming transcriptional regulation, single lab\",\n      \"pmids\": [\"41505216\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"JLP (SPAG9) is required for CD4+ T cell TCR-initiated proliferation, IL-2 production, and CD154 induction; JLP deficiency impairs Ca2+ influx and NF-AT activation but not MAPK, NF-κB, or AP-1 signaling pathways upon TCR stimulation.\",\n      \"method\": \"jlp-deficient mice CD4+ T cells, TCR stimulation, proliferation assay, cytokine measurement, Ca2+ flux assay, NF-AT reporter assay, pathway signaling Western blots\",\n      \"journal\": \"Molecular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO in primary T cells with specific pathway placement (Ca2+/NF-AT vs MAPK/NF-κB), single lab\",\n      \"pmids\": [\"28521278\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"SPAG9/JLP is a multifunctional scaffolding protein that tethers JNK and p38 MAPK signaling modules (along with upstream kinases MKK4/MEKK3) to transcription factors, interacts with kinesin-1 (via KLC1) and dynein motor complexes to regulate cargo-specific axonal and intracellular transport (including endosome-to-TGN trafficking with PIKfyve, lysosome positioning, and ARF6 delivery to the midbody during cytokinesis), recruits PLK1 to the centrosome for microtubule array formation and NF-κB nuclear import, interfaces with cell-surface receptors (Cdo, CD40) to couple extracellular signals to intracellular MAPK cascades, and also functions in sperm acrosomal localization and sperm-egg interaction.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"SPAG9 (JLP) is a multifunctional scaffolding protein that physically tethers stress-activated MAPK signaling modules — JNK and p38 — to upstream kinases (MKK4, MEKK3) and downstream transcription factors (Max, c-Myc), assembling pathway-specific signaling complexes [#0]. It binds JNK isoforms (JNK3 > JNK2 > JNK1) through a defined JNK-binding domain, with its leucine-zipper/coiled-coil and transmembrane regions governing dimerization and subcellular localization [#1]. A second major role is in microtubule-based transport: SPAG9 binds kinesin light chain 1 (KLC1) and kinesin-1 heavy chain to form motor-cargo complexes, with this interaction required for cytoplasmic localization and for ternary complex assembly with JNK [#3]. Through these motor linkages SPAG9, together with the redundant scaffold JSAP1, drives kinesin-1-dependent axonal transport whose loss causes cargo accumulation, JNK hyperactivation, and progressive neuronal death [#10, #12], delivers active ARF6 to the midbody during cytokinesis [#8], controls endosome-to-TGN trafficking of furin cargo via PIKfyve [#7], and positions lysosomes to support autophagosome-lysosome fusion [#19, #20]. SPAG9 couples cell-surface receptors to intracellular cascades, partnering with Cdo to enhance p38α/β activation during myoblast differentiation [#4] and mediating CD40 internalization in B lymphocytes through Rab5- and dynein-dependent vesicle transport [#11]. Upon extracellular stimulation it recruits PLK1 to the centrosome in a p38- and dynein-dependent manner to drive centrosome maturation, microtubule array formation, and microtubule-based NF-κB p65 nuclear translocation [#16]. In immune and developmental contexts SPAG9 also supports TCR-initiated Ca2+ influx and NF-AT activation in CD4+ T cells [#25] and retinoic-acid-induced endodermal differentiation downstream of Gα13 [#5]. SPAG9 localizes to the sperm acrosome and relocates to the equatorial segment after the acrosome reaction, and its disruption in mice causes male subfertility [#2, #6]. In disease, SPAG9 sustains MAPK-driven proliferation and invasion across multiple cancers [#13, #14, #18], and a SPAG9::JAK2 fusion from t(9;17) constitutively activates JAK2 to phosphorylate STAT1/3/5 and confer growth-factor-independent proliferation [#21]; its transcriptional repression by TGF-β1/β-catenin/LEF1 contributes to renal fibrosis [#24].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Established SPAG9/JLP as a MAPK scaffold, answering how JNK/p38 modules are spatially organized with upstream kinases and target transcription factors.\",\n      \"evidence\": \"Co-immunoprecipitation and interaction assays in transfected cells\",\n      \"pmids\": [\"12391307\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and structure of the assembled scaffold complex not resolved\", \"Physiological signals triggering complex assembly not defined\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Mapped the JNK-binding domain and the domains controlling dimerization/localization, defining how SPAG9 selectively engages JNK over p38α or ERK, and identified an acrosomal pool linked to sperm-egg binding.\",\n      \"evidence\": \"Deletion mutants, CD spectroscopy, immunofluorescence, and hemizona/egg penetration antibody-inhibition assays\",\n      \"pmids\": [\"15693750\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a single scaffold reconciles JNK-selectivity with reported p38 binding not resolved\", \"Molecular basis of acrosomal targeting unknown\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identified KLC1 as a kinesin-1 link, explaining how SPAG9 achieves cytoplasmic localization and couples motor transport to JNK signaling.\",\n      \"evidence\": \"Yeast two-hybrid, reciprocal co-IP, site-directed mutagenesis, and dominant-negative mislocalization assays\",\n      \"pmids\": [\"15987681\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cargoes transported by the SPAG9-KLC1-JNK complex not defined here\", \"Regulation of motor engagement unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Connected SPAG9 to cell-surface receptor signaling and developmental differentiation, showing it relays Cdo and Gα13 inputs into p38/JNK cascades.\",\n      \"evidence\": \"Co-IP, p38 activity assays, Cdo-/- primary myoblasts with MKK6 rescue, and siRNA in P19 cells\",\n      \"pmids\": [\"17074887\", \"16619266\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct vs indirect Cdo-SPAG9 contact regions not fully mapped\", \"Gα13-SPAG9 interaction is single-lab\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Demonstrated an in vivo physiological requirement, with male subfertility on Jlp disruption establishing relevance to spermatogenesis.\",\n      \"evidence\": \"Mouse gene knockout, immunohistochemistry, and fertility testing\",\n      \"pmids\": [\"18574703\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular defect in knockout spermatids not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Defined a cargo-specific trafficking function, showing SPAG9-PIKfyve is needed for microtubule-based endosome-to-TGN transport of furin but not microtubule-independent cargo.\",\n      \"evidence\": \"Yeast two-hybrid, pulldown, co-IP, siRNA depletion with ectopic rescue, and defined cargo-trafficking assays\",\n      \"pmids\": [\"19056739\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How cargo specificity is encoded not resolved\", \"Link to JNK scaffolding role unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Extended the motor-scaffold role to cytokinesis and UV stress, showing SPAG9 delivers ARF6 to the midbody and is required for UVB-induced p38-dependent keratinocyte apoptosis.\",\n      \"evidence\": \"Knockout MEFs/mice with binding-deficient mutant rescue, co-IP, cytokinesis assays, and p38 inhibitor treatment in skin\",\n      \"pmids\": [\"25130574\", \"24520900\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional redundancy with JSAP1 complicates single-protein attribution\", \"Upstream activator of midbody localization not defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Resolved the consequences of transport failure in neurons and the receptor-internalization role, showing kinesin-1-dependent axonal transport requires SPAG9/JSAP1 and that its loss drives dynein-mediated nuclear JNK relocalization and neuronal death, while CD40 internalization needs SPAG9 with Rab5/dynein.\",\n      \"evidence\": \"Conditional double-knockout mice, primary neuron rescue, kinesin-1 interaction assays, jlp-deficient B cells, and dynein inhibitor (ciliobrevin D)\",\n      \"pmids\": [\"25571974\", \"26320416\", \"25586186\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cargo selectivity among kinesin-1 substrates incompletely defined\", \"How the same scaffold engages both kinesin and dynein not mechanistically resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Linked SPAG9-JNK complex assembly and translocation to oncogenic signaling, showing LPA-stimulated perinuclear relocalization drives cancer proliferation and invasion, with ELK1 placed downstream in HCC.\",\n      \"evidence\": \"Co-IP, co-localization, shRNA/siRNA knockdown with ELK1 rescue, and xenograft models\",\n      \"pmids\": [\"27655714\", \"27042099\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab cancer-context findings\", \"Direct vs indirect ELK1 regulation not established\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established a cytoprotective JNK-dependent function, showing the JNK-binding domain is required for SPAG9-mediated protection against oxidative cell death.\",\n      \"evidence\": \"siRNA knockdown with wild-type vs JNK-binding-domain mutant rescue, viability and ROS assays\",\n      \"pmids\": [\"29753743\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab\", \"Mechanism linking JNK scaffolding to ROS control not defined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Unified the centrosome, lysosome, and pathway-specificity roles, showing SPAG9 tethers PLK1 to the centrosome for microtubule array formation and NF-κB nuclear import, positions lysosomes via kinesin-1 to enable autophagy, and engages an MKK3/p38 (not MKK6) axis in liver cancer.\",\n      \"evidence\": \"Live imaging, domain deletion, p38/dynein inhibitors, NF-κB transport assays, autophagy flux with mutant rescue, and co-IP with pathway Western blots\",\n      \"pmids\": [\"31803841\", \"32023558\", \"31787236\", \"30720101\", \"31497238\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How SPAG9 selects MKK3 over MKK6 not defined\", \"Prostate cancer MAPK link is Western-blot only (Low confidence)\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined a non-MAPK immune role, showing SPAG9 supports TCR-initiated Ca2+ influx and NF-AT activation independently of MAPK/NF-κB/AP-1.\",\n      \"evidence\": \"jlp-deficient CD4+ T cells, proliferation/cytokine assays, Ca2+ flux, NF-AT reporter, and pathway Western blots\",\n      \"pmids\": [\"28521278\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism coupling SPAG9 to Ca2+/NF-AT unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified a transcription-factor-tethering anti-EMT function, showing SPAG9 sequesters Foxk1 to relieve repression of N-cadherin and counteract TGF-β1-induced partial EMT.\",\n      \"evidence\": \"TGF-β1 treatment, JLP knockdown/overexpression, Foxk1 modulation, and N-cadherin expression assays\",\n      \"pmids\": [\"37013185\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct Foxk1-SPAG9 binding interface not mapped\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Revealed an oncogenic fusion mechanism, showing SPAG9::JAK2 constitutively activates JAK2 to drive STAT1/3/5 phosphorylation and a STAT1-BCL-2/MCL-1 survival axis conferring cytokine-independent growth.\",\n      \"evidence\": \"Ba/F3 reconstitution, IL-3-independent growth, kinase-domain mutagenesis, and ruxolitinib treatment\",\n      \"pmids\": [\"35879405\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Contribution of SPAG9 dimerization domains to fusion activation not dissected\", \"Prevalence in patients not addressed by these experiments\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Placed SPAG9 as a transcriptionally regulated brake on fibrosis-associated autophagy, showing TGF-β1/β-catenin/LEF1 represses SPAG9 to unleash autophagy and renal fibrosis, reversible by β-catenin/LEF1 inhibition.\",\n      \"evidence\": \"CKD patient tissue, murine fibrosis models, HK-2 cells, iCRT3/LEF1 silencing, and JLP expression/autophagy assays\",\n      \"pmids\": [\"41505216\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct LEF1 binding at the SPAG9 promoter not fully characterized\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single scaffold integrates seemingly contradictory roles — selective JNK vs p38 binding, simultaneous engagement of kinesin and dynein, and pro- vs anti-survival outputs — across cell types remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of the full SPAG9 scaffold or its cargo/motor interfaces\", \"Context-determinants selecting among partners not defined\", \"Relative contributions of SPAG9 vs redundant JSAP1 in vivo incompletely partitioned\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 3, 8]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [3, 8, 10, 19]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 4, 16]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [16]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [19, 20]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [7, 11]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [10, 16]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 4, 16]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [7, 8, 11, 19]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [19, 20, 24]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [11, 25]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 5]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [8]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [21]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"JNK3\", \"MKK4\", \"MEKK3\", \"KLC1\", \"PIKfyve\", \"ARF6\", \"PLK1\", \"Cdo\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":9,"faith_total":9,"faith_pct":100.0}}