{"gene":"TG_230180","gene_symbol":"GRA24","run_date":"2026-09-05T22:31:42","timeline":{"discoveries":[{"year":2013,"finding":"GRA24 is a dense granule protein secreted by Toxoplasma gondii that traffics from the parasitophorous vacuole to the host cell nucleus, where it triggers prolonged autophosphorylation and nuclear translocation of host p38α MAP kinase (MAPK14), leading to upregulation of transcription factors Egr-1 and c-Fos and synthesis of proinflammatory cytokines IL-12 and MCP-1. This activation kinetics is non-canonical, distinct from the standard MAPK signaling cascade.","method":"Dense granule protein identification, subcellular fractionation/imaging showing nuclear trafficking, co-immunoprecipitation of GRA24-p38α complex, cytokine measurements in infected cells","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP demonstrating complex formation, direct localization experiments, functional cytokine readouts, foundational paper replicated by multiple subsequent studies","pmids":["24043761"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"finding":"GRA24 is an intrinsically disordered protein that binds two p38α kinase molecules independently via an adapted kinase interaction motif (KIM), forming a highly stable complex that bypasses canonical MAPK activation and competes with cytoplasmic regulatory partners. The purified recombinant GRA24-p38α complex is the only factor required to activate p38α in vitro.","method":"Structural analysis of recombinant GRA24-human p38α complex, in vitro reconstitution, biochemical assays, crystallography/biophysical characterization","journal":"Structure (London, England : 1993)","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution of complex, structural determination, mutagenesis-level mechanistic insight into KIM motif, multiple orthogonal methods","pmids":["27889209"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2015,"finding":"TgASP5, a Golgi-resident aspartyl protease, is required for export of GRA24 beyond the parasitophorous vacuole to the host cell nucleus. In the absence of ASP5, GRA24 is retained within the vacuolar space. Notably, GRA24 maturation/cleavage per se is TgASP5-independent, indicating ASP5 plays a role in GRA24 export through a mechanism other than direct proteolytic processing of GRA24.","method":"ASP5 knockout parasites, immunofluorescence assay tracking GRA24 localization, epitope-tagged constructs","journal":"Cellular microbiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic knockout with direct localization readout, independently confirmed by two concurrent papers (PMID 26270241 and PMID 26473595)","pmids":["26270241","26473595"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"finding":"MYR1 is essential for transit of GRA24 to the host cell nucleus; deletion of MYR1 blocks GRA24 translocation across the parasitophorous vacuole membrane (PVM) into the host cell cytosol/nucleus. MYR1 localizes to the PVM and is cleaved by TgASP5, suggesting it is a component of the effector translocation machinery.","method":"MYR1 deletion mutants, immunofluorescence assay for GRA24 nuclear localization, whole-genome sequencing of mutants, subcellular fractionation","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic knockout with direct localization readout, epistasis linking MYR1 to GRA24 transport, replicated by subsequent translocon studies","pmids":["26838724"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"RH"}]},{"year":2018,"finding":"Novel proteins MYR2 and MYR3 are components of a complex with MYR1 at the PVM that is essential for translocation of GRA24 (and GRA16) into the host cell. MYR3 stably associates with MYR1 whose N- and C-terminal fragments are disulfide-linked. Fusion of GRA24 to a structured DHFR domain blocks its transport, consistent with a translocon-based unfolding mechanism.","method":"Genetic screen for c-Myc up-regulation defects, MYR2/MYR3 deletion, co-immunoprecipitation/co-localization of MYR1/2/3 at PVM, GFP-tracking of GRA24 nuclear localization, DHFR fusion experiments","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP showing MYR1/MYR3 association, direct localization, genetic epistasis with functional GRA24 transport readout, multiple orthogonal methods","pmids":["29357375"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2015,"finding":"Yeast three-hybrid analysis identified GRA24 (also named TgBRADIN) as a parasite protein that interacts with the small-molecule Compound 2 (an imidazopyridine). Disruption of the GRA24 gene leads to enhanced bradyzoite differentiation, identifying GRA24 as a negative regulator of bradyzoite/stage conversion.","method":"Yeast three-hybrid screen, GRA24 gene disruption/knockout, differentiation assays measuring bradyzoite stage conversion","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast three-hybrid plus gene disruption with phenotypic readout, single lab, functional consequence established but molecular mechanism of differentiation suppression not fully elucidated","pmids":["25789621"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2018,"finding":"GRA24 (and GRA16) are not secreted beyond the cyst wall membrane in bradyzoite tissue cysts. Using tetracycline-inducible expression of HA-tagged GRA16 and GRA24 after stage conversion, these proteins were retained within the cyst and did not traffic to the host cell nucleus, demonstrating that PVM effector export is stage-specific and absent in the bradyzoite/cyst stage.","method":"Tetracycline-inducible expression system, HA-tagged GRA16 and GRA24 in bradyzoite cysts, immunofluorescence assay","journal":"Frontiers in cellular and infection microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct experimental localization using inducible tagged constructs in bradyzoites, single lab, negative result but informative for stage-specificity","pmids":["30406043"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"finding":"GRA24 drives a sustained, non-canonical Egr-1 response in dendritic cells through p38 MAPK signaling. The extended-phase Egr-1 nuclear accumulation in infected DCs was dramatically reduced upon challenge with GRA24-deficient T. gondii parasites, and this response was MyD88-independent. GRA24-dependent Egr-1 upregulation suppressed DC maturation (reduced CD40, CD80 expression) and modulated IL-12 and IL-2 responses.","method":"GRA24 knockout parasites, Egr-1 siRNA silencing in primary DCs, pharmacological p38 inhibition, immunofluorescence for nuclear Egr-1, cytokine measurements, flow cytometry for surface markers","journal":"Frontiers in cellular and infection microbiology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout combined with pharmacological inhibition and siRNA knockdown, multiple orthogonal functional readouts, direct nuclear localization experiments","pmids":["31681626"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"finding":"GRA24 drives a late-phase p38 MAPK phosphorylation response in macrophages that is MyD88-independent and does not require active parasite proliferation, leading to IL-12 production. This GRA24-dependent p38 MAPK activation promotes a host-protective Th1 response independently of the TLR/MyD88 cascade.","method":"GRA24 knockout in uracil auxotrophic cps1-1 strain, bone marrow-derived macrophage infection assays, IL-12-YFP reporter mice, p38 phosphorylation assays, cytokine profiling in MyD88-/- mice","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean GRA24 deletion, multiple host genetic backgrounds (MyD88+/+ and MyD88-/-), in vivo and in vitro readouts, multiple orthogonal methods","pmids":["32413093"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"cps1-1 (Type I uracil auxotroph)"}]},{"year":2020,"finding":"GRA15 and GRA24 synergistically activate the innate immune response in TLR11-deficient mice and human macrophages. GRA15 and GRA24 are major drivers of IL-12, IL-18, and IL-1β secretion in human macrophages, which subsequently activates IFNγ secretion by PBMCs to control parasite proliferation.","method":"GRA15 and GRA24 single and double knockout parasites, infection of Tlr11-/- mice and human macrophages, cytokine ELISA (IL-12, IL-18, IL-1β, IFNγ), PBMC co-culture assays","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic knockouts (single and double), multiple host genetic backgrounds, multiple orthogonal functional readouts (in vivo and in vitro)","pmids":["32453782"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"finding":"GRA24 promotes CCR7-mediated macrophage chemotaxis (hypermigration) via p38 MAPK signaling, enabling dissemination of T. gondii type II lineage. GRA24 and GRA15 act cooperatively: GRA15 activates NF-κB while GRA24 activates p38 MAPK, and GRA28 boosts chromatin accessibility at the Ccr7 gene locus to amplify CCR7 expression. In vivo, macrophages infected with wild-type T. gondii outcompeted GRA15/24 double-mutant-infected macrophages in migrating to secondary organs.","method":"GRA15/GRA24 single and double knockout parasites, CCR7 expression and chemotaxis assays, p38 MAPK and NF-κB signaling inhibitors, ATAC-seq for chromatin accessibility, adoptive transfer in vivo dissemination assay","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic knockouts, in vitro signaling assays, ATAC-seq, and in vivo adoptive transfer experiments; multiple orthogonal methods across two publications (peer-reviewed and preprint)","pmids":["39207098","38370679"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"type II"}]},{"year":2024,"finding":"GRA24 is one of four nuclear-targeted effectors (TgIST, GRA16, GRA24, GRA28) necessary for a complete block of IFNγ-driven host cell death and parasite premature egress in human (but not murine) cells. GRA24's protective action in this context depends on its interaction with p38 MAPK, and this function is MYR translocon-dependent.","method":"Genome-wide loss-of-function screens in IFNγ-stimulated human cells, combinatorial GRA knockout parasites, MYR translocon inhibition, cell death and egress assays, IFNγ-driven transcription assays","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide screen followed by targeted knockouts, multiple human cell lines, functional rescue experiments, p38 MAPK interaction mechanistically confirmed","pmids":["39292011"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"finding":"GRA24-dependent p38 MAPK signaling in host brain endothelial cells induces elevated transcription and secretion of TIMP1, which promotes non-disruptive T. gondii transmigration across polarized brain endothelial (bEnd.3) and intestinal epithelial (Caco-2) monolayers without disrupting barrier integrity. Pharmacological inhibition of p38 and use of GRA24 knockout parasites abolished TIMP1 induction.","method":"GRA24 knockout T. gondii, pharmacological p38 MAPK inhibition, transwell transmigration assays, TIMP1 ELISA and transcription analysis, barrier integrity measurements (TEER), recombinant TIMP1 supplementation","journal":"mSphere","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout and pharmacological inhibition with functional barrier transmigration readout, single lab, novel function identified","pmids":["40265926"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2026,"finding":"GRA24 is one of eight MYR1-dependent dense granule effectors (IST, NSM, HCE1/TEEGR, GRA16, GRA18, GRA24, GRA28, GRA84) that function cooperatively to activate the non-canonical NF-κB pathway (RelB/p52) in host cells. This activation involves parasite-driven depletion of TRAF3, stabilization of NF-κB-inducing kinase (NIK), phosphorylation and processing of p100 to p52. No single effector alone accounts for this phenotype.","method":"Combinatorial GRA knockout parasites (panel-based), immunoblotting for RelB/p52/p100/NIK/TRAF3, nuclear fractionation, genome-wide loss-of-function screening","journal":"bioRxiv : the preprint server for biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — combinatorial knockout panel with mechanistic pathway markers, preprint not yet peer-reviewed, single lab","pmids":["41959509"],"is_preprint":true,"attribution":[{"species":"Toxoplasma gondii","strain":"type I; type II"}]},{"year":2016,"finding":"Deletion of GRA24 in uracil auxotrophic T. gondii strains severely abrogated antitumor immune responses, demonstrating that GRA24's trafficking to the host cell nucleus is functionally required for activating host immunity (CD8α+ dendritic cells, IL-12/IFNγ Th1 axis) that controls tumor growth.","method":"Targeted deletion of GRA24 in Δku80 vaccine strain, mouse tumor model, immune cell profiling by flow cytometry, cytokine measurements","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean gene deletion with defined in vivo phenotypic readout, single lab, functional role confirmed","pmids":["27447180"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"type I Δku80"}]},{"year":2024,"finding":"GRA24-induced IL-12 production acts as a virulence factor in T cell-deficient (Tcrb-/-) and IL-10-deficient mouse hosts by driving fatal cytokine storm; resistance was restored in Tcrb-/- mice by monoclonal depletion of IL-12p40, demonstrating that GRA24-driven IL-12 underlies immunopathology in the absence of regulatory T cell responses.","method":"GRA24 knockout uracil auxotroph parasites, Tcrb-/- and IL-10-/- mice, antibody depletion of IL-12p40, survival assays, cytokine profiling","journal":"mBio","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic knockout with multiple mouse genetic backgrounds and antibody depletion rescue, single lab, multiple functional readouts","pmids":["39440975"],"is_preprint":false,"attribution":[{"species":"Toxoplasma gondii","strain":"type I uracil auxotroph (OMP)"}]}],"current_model":"GRA24 is a Toxoplasma gondii dense granule protein secreted into the parasitophorous vacuole that is exported beyond the vacuole membrane via the MYR1/MYR2/MYR3 translocon complex (facilitated by TgASP5) and traffics to the host cell nucleus, where the intrinsically disordered protein binds two p38α MAPK molecules via an adapted kinase interaction motif (KIM) to bypass canonical MAPK activation, driving sustained autophosphorylation and nuclear translocation of p38α, upregulation of Egr-1 and c-Fos, production of proinflammatory cytokines (IL-12, IL-1β, MCP-1), CCR7-mediated macrophage hypermigration for parasite dissemination, TIMP1-mediated barrier transmigration, and suppression of IFNγ-driven host cell death; GRA24 additionally acts as a negative regulator of bradyzoite differentiation and, in concert with other GRA effectors, contributes to non-canonical NF-κB pathway activation in infected host cells."},"narrative":{"mechanistic_narrative":"GRA24 is a Toxoplasma gondii dense granule effector that is secreted into the parasitophorous vacuole, exported into the host cell, and traffics to the host nucleus, where it functions as a non-canonical activator of host p38α MAP kinase to reprogram infected-cell signaling and drive proinflammatory responses [PMID:24043761]. GRA24 is intrinsically disordered and binds two p38α molecules independently through an adapted kinase interaction motif (KIM), forming a stable complex that bypasses the canonical MAPK cascade; the recombinant GRA24–p38α complex is the sole factor required to activate p38α in vitro [PMID:27889209]. Downstream, sustained p38α autophosphorylation and nuclear translocation drive Egr-1 and c-Fos upregulation and synthesis of proinflammatory cytokines including IL-12 and MCP-1 [PMID:24043761], a response that is MyD88-independent and does not require active parasite proliferation [PMID:32413093]. Export of GRA24 beyond the vacuole requires the Golgi aspartyl protease TgASP5 [PMID:26270241, PMID:26473595] and the PVM-resident MYR1/MYR2/MYR3 translocon complex [PMID:26838724, PMID:29357375], and this export is stage-specific, being absent in bradyzoite cysts [PMID:30406043]. Functionally, GRA24 acts together with other GRA effectors to shape host immunity: it cooperates with GRA15 (and GRA28) to drive CCR7-mediated macrophage hypermigration and parasite dissemination [PMID:39207098, PMID:38370679], synergizes with GRA15 in IL-12/IL-18/IL-1β secretion and downstream IFNγ control [PMID:32453782], and contributes alongside TgIST, GRA16 and GRA28 to blocking IFNγ-driven host cell death via its p38 interaction [PMID:39292011]. Independently of its host-directed role, GRA24 (TgBRADIN) acts within the parasite as a negative regulator of bradyzoite differentiation [PMID:25789621].","teleology":[{"year":2013,"claim":"Established that a parasite dense granule protein could reach and reprogram the host nucleus, defining GRA24 as an exported effector that non-canonically activates host p38α MAPK and its transcriptional output.","evidence":"Dense granule protein identification, nuclear trafficking imaging, GRA24–p38α co-immunoprecipitation and cytokine readouts in infected cells","pmids":["24043761"],"confidence":"High","gaps":["Structural basis of p38α binding not resolved","Mechanism of vacuolar-to-nuclear export not defined"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2015,"claim":"Identified GRA24 (TgBRADIN) as a parasite-intrinsic negative regulator of stage conversion, distinguishing a role in bradyzoite differentiation from its host-directed signaling function.","evidence":"Yeast three-hybrid screen against Compound 2 plus GRA24 gene disruption with bradyzoite differentiation assays","pmids":["25789621"],"confidence":"Medium","gaps":["Molecular mechanism of differentiation suppression not elucidated","Relationship to host p38α signaling role unclear"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2015,"claim":"Resolved a step in the export route by showing that the Golgi aspartyl protease TgASP5 is required for GRA24 export beyond the vacuole, and that this occurs independently of direct proteolytic processing of GRA24.","evidence":"ASP5 knockout parasites with immunofluorescence tracking of GRA24 localization","pmids":["26270241","26473595"],"confidence":"High","gaps":["How ASP5 promotes export without cleaving GRA24 not defined","Substrate/motif requirements on GRA24 unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"claim":"Provided the structural and biochemical mechanism, showing GRA24 is intrinsically disordered and captures two p38α molecules via an adapted KIM to form the minimal, sufficient activating complex.","evidence":"Structural analysis, in vitro reconstitution and biophysical characterization of recombinant GRA24–human p38α complex","pmids":["27889209"],"confidence":"High","gaps":["How the disordered protein bypasses upstream MAPK kinases mechanistically","In vivo stoichiometry and turnover of the complex not established"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2016,"claim":"Defined MYR1 as an essential PVM translocation component for GRA24 delivery, linking effector export machinery to GRA24 nuclear trafficking.","evidence":"MYR1 deletion mutants with immunofluorescence for GRA24 nuclear localization and subcellular fractionation","pmids":["26838724"],"confidence":"High","gaps":["Composition of the full translocon not yet known","Direct interaction between GRA24 and MYR1 not shown"],"attribution":[{"species":"Toxoplasma gondii","strain":"RH"}]},{"year":2016,"claim":"Demonstrated the in vivo functional consequence of GRA24 export, showing its nuclear trafficking is required to activate host antitumor immunity via CD8α+ DCs and the IL-12/IFNγ Th1 axis.","evidence":"Targeted GRA24 deletion in Δku80 vaccine strain, mouse tumor model, immune cell profiling and cytokine measurement","pmids":["27447180"],"confidence":"Medium","gaps":["Direct causal chain from p38α to specific immune output not dissected","Contribution relative to other effectors not quantified"],"attribution":[{"species":"Toxoplasma gondii","strain":"type I Δku80"}]},{"year":2018,"claim":"Extended the translocon model by identifying MYR2 and MYR3 as MYR1-associated PVM components required for GRA24 translocation, and provided evidence for a translocon-based unfolding mechanism.","evidence":"Genetic screen, MYR2/MYR3 deletion, reciprocal Co-IP/co-localization at PVM, GFP tracking, and DHFR-fusion transport-block experiments","pmids":["29357375"],"confidence":"High","gaps":["Molecular architecture of the translocon pore not resolved","Energetics/driving force of unfolding-coupled transport unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2018,"claim":"Established stage-specificity of GRA24 export, showing it is not secreted beyond the cyst wall in bradyzoites, restricting its host-nuclear function to the tachyzoite stage.","evidence":"Tetracycline-inducible HA-tagged GRA24 expression in bradyzoite cysts with immunofluorescence localization","pmids":["30406043"],"confidence":"Medium","gaps":["Mechanism silencing export in bradyzoites unknown","Whether translocon is downregulated or effector retained is not distinguished"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2019,"claim":"Connected GRA24–p38 signaling to dendritic cell reprogramming, showing GRA24 drives sustained Egr-1 to suppress DC maturation and modulate cytokine output independently of MyD88.","evidence":"GRA24 knockout parasites, Egr-1 siRNA, pharmacological p38 inhibition, nuclear Egr-1 imaging, cytokine and surface-marker flow cytometry in primary DCs","pmids":["31681626"],"confidence":"High","gaps":["Downstream transcriptional targets of Egr-1 not mapped","Net effect on protective vs suppressive immunity not resolved"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2020,"claim":"Showed that GRA24-driven late-phase p38 activation in macrophages promotes a host-protective Th1/IL-12 response independent of TLR/MyD88 and parasite replication.","evidence":"GRA24 knockout in cps1-1 strain, BMDM infection, IL-12-YFP reporter mice, p38 phosphorylation assays, cytokine profiling in MyD88-/- mice","pmids":["32413093"],"confidence":"High","gaps":["How p38 activation kinetics translate to sustained IL-12 transcription not detailed","In vivo cellular source of protective cytokines not fully defined"],"attribution":[{"species":"Toxoplasma gondii","strain":"cps1-1 (Type I uracil auxotroph)"}]},{"year":2020,"claim":"Placed GRA24 within a cooperative effector program, showing synergy with GRA15 in driving IL-12/IL-18/IL-1β secretion and downstream IFNγ-mediated control of the parasite.","evidence":"GRA15/GRA24 single and double knockout parasites, Tlr11-/- mice and human macrophages, cytokine ELISA and PBMC co-culture","pmids":["32453782"],"confidence":"High","gaps":["Molecular convergence of GRA15 (NF-κB) and GRA24 (p38) pathways not resolved","Human-specific vs murine differences not fully explored"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"claim":"Defined a dissemination mechanism in which GRA24-driven p38, cooperating with GRA15 (NF-κB) and GRA28 (chromatin accessibility at Ccr7), promotes CCR7-mediated macrophage hypermigration to secondary organs.","evidence":"GRA15/GRA24 single and double knockouts, CCR7 and chemotaxis assays, p38/NF-κB inhibitors, ATAC-seq, in vivo adoptive transfer dissemination","pmids":["39207098","38370679"],"confidence":"High","gaps":["Direct transcriptional target of GRA24-p38 at Ccr7 not shown","Relative contribution of each effector in vivo not quantified"],"attribution":[{"species":"Toxoplasma gondii","strain":"type II"}]},{"year":2024,"claim":"Identified a barrier-crossing function whereby GRA24-p38 signaling in host endothelial/epithelial cells induces TIMP1 to enable non-disruptive parasite transmigration.","evidence":"GRA24 knockout parasites, p38 inhibition, transwell transmigration, TIMP1 ELISA/transcription, TEER integrity, recombinant TIMP1 rescue","pmids":["40265926"],"confidence":"Medium","gaps":["Mechanism by which TIMP1 facilitates transmigration not defined","Single-lab observation of this specific function"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"claim":"Positioned GRA24 among nuclear effectors required to block IFNγ-driven host cell death, with its protective action depending on p38 interaction and MYR translocon delivery in human but not murine cells.","evidence":"Genome-wide loss-of-function screens in IFNγ-stimulated human cells, combinatorial GRA knockouts, MYR inhibition, cell death and egress assays","pmids":["39292011"],"confidence":"High","gaps":["Molecular basis of human vs murine difference not resolved","How p38 activation suppresses IFNγ-driven death not mechanistically detailed"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]},{"year":2024,"claim":"Revealed a double-edged consequence of GRA24 signaling, showing GRA24-induced IL-12 acts as a virulence factor causing fatal cytokine storm in T cell- or IL-10-deficient hosts.","evidence":"GRA24 knockout uracil auxotrophs, Tcrb-/- and IL-10-/- mice, IL-12p40 antibody depletion rescue, survival and cytokine profiling","pmids":["39440975"],"confidence":"Medium","gaps":["Threshold determining protective vs pathogenic IL-12 not defined","Applicability to wild-type immunocompetent hosts unclear"],"attribution":[{"species":"Toxoplasma gondii","strain":"type I uracil auxotroph (OMP)"}]},{"year":2026,"claim":"Placed GRA24 within a cooperative MYR1-dependent effector set that activates the non-canonical NF-κB (RelB/p52) pathway via TRAF3 depletion, NIK stabilization, and p100 processing, with no single effector sufficient.","evidence":"Combinatorial GRA knockout panels, immunoblotting for RelB/p52/p100/NIK/TRAF3, nuclear fractionation, genome-wide screening (preprint)","pmids":["41959509"],"confidence":"Medium","gaps":["Specific contribution of GRA24 to this pathway not isolated","Preprint not yet peer-reviewed"],"attribution":[{"species":"Toxoplasma gondii","strain":"type I"},{"species":"Toxoplasma gondii","strain":"type II"}]},{"year":null,"claim":"How GRA24 engages the MYR translocon for transport and how its p38α activation is integrated with parallel GRA-driven NF-κB signaling to yield distinct host outcomes remain open.","evidence":"No direct evidence in the available corpus","pmids":[],"confidence":"Low","gaps":["Direct GRA24–translocon interaction not demonstrated","Convergence of p38 and NF-κB branches on shared host targets not resolved","Mechanism of bradyzoite differentiation suppression unknown"],"attribution":[{"species":"Toxoplasma gondii","strain":""}]}],"mechanism_profile":{"molecular_activity":[],"localization":[],"pathway":[],"complexes":["GRA24–p38α (MAPK14) complex (two p38α per GRA24 via adapted KIM)"],"partners":["HOST P38Α MAPK (MAPK14)","TGASP5 (EXPORT DEPENDENCY)","MYR1 (TRANSLOCATION DEPENDENCY)","MYR2 (TRANSLOCATION DEPENDENCY)","MYR3 (TRANSLOCATION DEPENDENCY)","GRA15 (FUNCTIONAL COOPERATION)","GRA28 (FUNCTIONAL COOPERATION)"],"other_free_text":["Intrinsically disordered dense granule effector exported beyond the parasitophorous vacuole membrane","Drives sustained/non-canonical p38α activation independent of MyD88 and independent of active parasite proliferation","Downstream host transcriptional output includes Egr-1, c-Fos, IL-12, MCP-1, and TIMP1","Export is stage-specific: active in tachyzoites, absent in bradyzoite cysts","Parasite-intrinsic role (as TgBRADIN) as negative regulator of bradyzoite differentiation","GRA24-driven IL-12 can be host-protective or pathogenic (cytokine storm) depending on host immune context"]},"strain_conflicts":[]},"prefetch_data":{},"corpus_meta":[{"pmid":"24043761","id":"PMC_24043761","title":"A Toxoplasma dense granule protein, GRA24, modulates the early immune response to infection by promoting a direct and sustained host p38 MAPK activation.","date":"2013","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/24043761","citation_count":219,"is_preprint":false},{"pmid":"26838724","id":"PMC_26838724","title":"A Novel Secreted Protein, MYR1, Is Central to Toxoplasma's Manipulation of Host Cells.","date":"2016","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/26838724","citation_count":133,"is_preprint":false},{"pmid":"26473595","id":"PMC_26473595","title":"Fundamental Roles of the Golgi-Associated Toxoplasma Aspartyl Protease, ASP5, at the Host-Parasite Interface.","date":"2015","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/26473595","citation_count":95,"is_preprint":false},{"pmid":"26270241","id":"PMC_26270241","title":"The aspartyl protease TgASP5 mediates the export of the Toxoplasma GRA16 and GRA24 effectors into host cells.","date":"2015","source":"Cellular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/26270241","citation_count":84,"is_preprint":false},{"pmid":"29357375","id":"PMC_29357375","title":"Identification of a novel protein complex essential for effector translocation across the parasitophorous vacuole membrane of Toxoplasma gondii.","date":"2018","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/29357375","citation_count":75,"is_preprint":false},{"pmid":"24373221","id":"PMC_24373221","title":"Toxoplasma exports dense granule proteins beyond the vacuole to the host cell nucleus and rewires the host genome expression.","date":"2014","source":"Cellular microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/24373221","citation_count":54,"is_preprint":false},{"pmid":"27447180","id":"PMC_27447180","title":"Secretion of Rhoptry and Dense Granule Effector Proteins by Nonreplicating Toxoplasma gondii Uracil Auxotrophs Controls the Development of Antitumor Immunity.","date":"2016","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/27447180","citation_count":47,"is_preprint":false},{"pmid":"32453782","id":"PMC_32453782","title":"Toxoplasma  GRA15 and GRA24 are important activators of the host innate immune response in the absence of TLR11.","date":"2020","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/32453782","citation_count":42,"is_preprint":false},{"pmid":"27889209","id":"PMC_27889209","title":"Structural Basis for the Subversion of MAP Kinase Signaling by an Intrinsically Disordered Parasite Secreted Agonist.","date":"2016","source":"Structure (London, England : 1993)","url":"https://pubmed.ncbi.nlm.nih.gov/27889209","citation_count":38,"is_preprint":false},{"pmid":"32413093","id":"PMC_32413093","title":"Toxoplasma gondii dense granule protein GRA24 drives MyD88-independent p38 MAPK activation, IL-12 production and induction of protective immunity.","date":"2020","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/32413093","citation_count":35,"is_preprint":false},{"pmid":"30894865","id":"PMC_30894865","title":"GRA24-Based DNA Vaccine Prolongs Survival in Mice Challenged With a Virulent Toxoplasma gondii Strain.","date":"2019","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/30894865","citation_count":33,"is_preprint":false},{"pmid":"31681626","id":"PMC_31681626","title":"Sustained Egr-1 Response via p38 MAP Kinase Signaling Modulates Early Immune Responses of Dendritic Cells Parasitized by Toxoplasma gondii.","date":"2019","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/31681626","citation_count":22,"is_preprint":false},{"pmid":"30406043","id":"PMC_30406043","title":"Toxoplasma Does Not Secrete the GRA16 and GRA24 Effectors Beyond the Parasitophorous Vacuole Membrane of Tissue Cysts.","date":"2018","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/30406043","citation_count":21,"is_preprint":false},{"pmid":"31535970","id":"PMC_31535970","title":"Evaluation of immune protection against Toxoplasma gondii infection in mice induced by a multi-antigenic DNA vaccine containing TgGRA24, TgGRA25 and TgMIC6.","date":"2019","source":"Parasite (Paris, France)","url":"https://pubmed.ncbi.nlm.nih.gov/31535970","citation_count":20,"is_preprint":false},{"pmid":"39207098","id":"PMC_39207098","title":"Hypermigration of macrophages through the concerted action of GRA effectors on NF-κB/p38 signaling and host chromatin accessibility potentiates Toxoplasma dissemination.","date":"2024","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/39207098","citation_count":13,"is_preprint":false},{"pmid":"25789621","id":"PMC_25789621","title":"Yeast three-hybrid screen identifies TgBRADIN/GRA24 as a negative regulator of Toxoplasma gondii bradyzoite differentiation.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25789621","citation_count":10,"is_preprint":false},{"pmid":"39292011","id":"PMC_39292011","title":"A combination of four Toxoplasma gondii nuclear-targeted effectors protects against interferon gamma-driven human host cell death.","date":"2024","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/39292011","citation_count":8,"is_preprint":false},{"pmid":"40265926","id":"PMC_40265926","title":"TIMP1 secretion induced by Toxoplasma effector GRA24 via p38 MAPK signaling promotes non-disruptive parasite translocation across polarized brain endothelial monolayers.","date":"2025","source":"mSphere","url":"https://pubmed.ncbi.nlm.nih.gov/40265926","citation_count":5,"is_preprint":false},{"pmid":"39440975","id":"PMC_39440975","title":"T lymphocyte-dependent IL-10 down-regulates a cytokine storm driven by Toxoplasma gondii GRA24.","date":"2024","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/39440975","citation_count":4,"is_preprint":false},{"pmid":"38370679","id":"PMC_38370679","title":"Hypermigration of macrophages through the concerted action of GRA effectors on NF-κB/p38 signaling and host chromatin accessibility potentiates Toxoplasma dissemination.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38370679","citation_count":1,"is_preprint":false},{"pmid":"38234811","id":"PMC_38234811","title":"A Combination of Four Nuclear Targeted Effectors Protects Toxoplasma Against Interferon Gamma Driven Human Host Cell Death During Acute Infection.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38234811","citation_count":1,"is_preprint":false},{"pmid":"38952314","id":"PMC_38952314","title":"[Preparation and preliminary application of the polyclonal antibody against Toxoplasma gondii dense granule protein 24].","date":"2024","source":"Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control","url":"https://pubmed.ncbi.nlm.nih.gov/38952314","citation_count":0,"is_preprint":false},{"pmid":"41959509","id":"PMC_41959509","title":"A Suite of Eight Toxoplasma gondii Effectors Cooperates to Activate the Non-canonical NF-κB Pathway.","date":"2026","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/41959509","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":15440,"output_tokens":5138,"usd":0.061695,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11919,"output_tokens":6420,"usd":0.110048,"stage2_stop_reason":"end_turn"},"total_usd":0.171743,"stage1_batch_id":"msgbatch_01XFXJD6iZecXyWn9uU5Qom9","stage2_batch_id":"msgbatch_013GToKgs4AKSCRE4yvueviH","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2013,\n      \"finding\": \"GRA24 is a dense granule protein secreted by Toxoplasma gondii that traffics from the parasitophorous vacuole to the host cell nucleus, where it triggers prolonged autophosphorylation and nuclear translocation of host p38α MAP kinase (MAPK14), leading to upregulation of transcription factors Egr-1 and c-Fos and synthesis of proinflammatory cytokines IL-12 and MCP-1. This activation kinetics is non-canonical, distinct from the standard MAPK signaling cascade.\",\n      \"method\": \"Dense granule protein identification, subcellular fractionation/imaging showing nuclear trafficking, co-immunoprecipitation of GRA24-p38α complex, cytokine measurements in infected cells\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP demonstrating complex formation, direct localization experiments, functional cytokine readouts, foundational paper replicated by multiple subsequent studies\",\n      \"pmids\": [\"24043761\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"GRA24 is an intrinsically disordered protein that binds two p38α kinase molecules independently via an adapted kinase interaction motif (KIM), forming a highly stable complex that bypasses canonical MAPK activation and competes with cytoplasmic regulatory partners. The purified recombinant GRA24-p38α complex is the only factor required to activate p38α in vitro.\",\n      \"method\": \"Structural analysis of recombinant GRA24-human p38α complex, in vitro reconstitution, biochemical assays, crystallography/biophysical characterization\",\n      \"journal\": \"Structure (London, England : 1993)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution of complex, structural determination, mutagenesis-level mechanistic insight into KIM motif, multiple orthogonal methods\",\n      \"pmids\": [\"27889209\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TgASP5, a Golgi-resident aspartyl protease, is required for export of GRA24 beyond the parasitophorous vacuole to the host cell nucleus. In the absence of ASP5, GRA24 is retained within the vacuolar space. Notably, GRA24 maturation/cleavage per se is TgASP5-independent, indicating ASP5 plays a role in GRA24 export through a mechanism other than direct proteolytic processing of GRA24.\",\n      \"method\": \"ASP5 knockout parasites, immunofluorescence assay tracking GRA24 localization, epitope-tagged constructs\",\n      \"journal\": \"Cellular microbiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic knockout with direct localization readout, independently confirmed by two concurrent papers (PMID 26270241 and PMID 26473595)\",\n      \"pmids\": [\"26270241\", \"26473595\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MYR1 is essential for transit of GRA24 to the host cell nucleus; deletion of MYR1 blocks GRA24 translocation across the parasitophorous vacuole membrane (PVM) into the host cell cytosol/nucleus. MYR1 localizes to the PVM and is cleaved by TgASP5, suggesting it is a component of the effector translocation machinery.\",\n      \"method\": \"MYR1 deletion mutants, immunofluorescence assay for GRA24 nuclear localization, whole-genome sequencing of mutants, subcellular fractionation\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic knockout with direct localization readout, epistasis linking MYR1 to GRA24 transport, replicated by subsequent translocon studies\",\n      \"pmids\": [\"26838724\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}]\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Novel proteins MYR2 and MYR3 are components of a complex with MYR1 at the PVM that is essential for translocation of GRA24 (and GRA16) into the host cell. MYR3 stably associates with MYR1 whose N- and C-terminal fragments are disulfide-linked. Fusion of GRA24 to a structured DHFR domain blocks its transport, consistent with a translocon-based unfolding mechanism.\",\n      \"method\": \"Genetic screen for c-Myc up-regulation defects, MYR2/MYR3 deletion, co-immunoprecipitation/co-localization of MYR1/2/3 at PVM, GFP-tracking of GRA24 nuclear localization, DHFR fusion experiments\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP showing MYR1/MYR3 association, direct localization, genetic epistasis with functional GRA24 transport readout, multiple orthogonal methods\",\n      \"pmids\": [\"29357375\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Yeast three-hybrid analysis identified GRA24 (also named TgBRADIN) as a parasite protein that interacts with the small-molecule Compound 2 (an imidazopyridine). Disruption of the GRA24 gene leads to enhanced bradyzoite differentiation, identifying GRA24 as a negative regulator of bradyzoite/stage conversion.\",\n      \"method\": \"Yeast three-hybrid screen, GRA24 gene disruption/knockout, differentiation assays measuring bradyzoite stage conversion\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast three-hybrid plus gene disruption with phenotypic readout, single lab, functional consequence established but molecular mechanism of differentiation suppression not fully elucidated\",\n      \"pmids\": [\"25789621\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"GRA24 (and GRA16) are not secreted beyond the cyst wall membrane in bradyzoite tissue cysts. Using tetracycline-inducible expression of HA-tagged GRA16 and GRA24 after stage conversion, these proteins were retained within the cyst and did not traffic to the host cell nucleus, demonstrating that PVM effector export is stage-specific and absent in the bradyzoite/cyst stage.\",\n      \"method\": \"Tetracycline-inducible expression system, HA-tagged GRA16 and GRA24 in bradyzoite cysts, immunofluorescence assay\",\n      \"journal\": \"Frontiers in cellular and infection microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct experimental localization using inducible tagged constructs in bradyzoites, single lab, negative result but informative for stage-specificity\",\n      \"pmids\": [\"30406043\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"GRA24 drives a sustained, non-canonical Egr-1 response in dendritic cells through p38 MAPK signaling. The extended-phase Egr-1 nuclear accumulation in infected DCs was dramatically reduced upon challenge with GRA24-deficient T. gondii parasites, and this response was MyD88-independent. GRA24-dependent Egr-1 upregulation suppressed DC maturation (reduced CD40, CD80 expression) and modulated IL-12 and IL-2 responses.\",\n      \"method\": \"GRA24 knockout parasites, Egr-1 siRNA silencing in primary DCs, pharmacological p38 inhibition, immunofluorescence for nuclear Egr-1, cytokine measurements, flow cytometry for surface markers\",\n      \"journal\": \"Frontiers in cellular and infection microbiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout combined with pharmacological inhibition and siRNA knockdown, multiple orthogonal functional readouts, direct nuclear localization experiments\",\n      \"pmids\": [\"31681626\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"GRA24 drives a late-phase p38 MAPK phosphorylation response in macrophages that is MyD88-independent and does not require active parasite proliferation, leading to IL-12 production. This GRA24-dependent p38 MAPK activation promotes a host-protective Th1 response independently of the TLR/MyD88 cascade.\",\n      \"method\": \"GRA24 knockout in uracil auxotrophic cps1-1 strain, bone marrow-derived macrophage infection assays, IL-12-YFP reporter mice, p38 phosphorylation assays, cytokine profiling in MyD88-/- mice\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean GRA24 deletion, multiple host genetic backgrounds (MyD88+/+ and MyD88-/-), in vivo and in vitro readouts, multiple orthogonal methods\",\n      \"pmids\": [\"32413093\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"cps1-1 (Type I uracil auxotroph)\"}]\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"GRA15 and GRA24 synergistically activate the innate immune response in TLR11-deficient mice and human macrophages. GRA15 and GRA24 are major drivers of IL-12, IL-18, and IL-1β secretion in human macrophages, which subsequently activates IFNγ secretion by PBMCs to control parasite proliferation.\",\n      \"method\": \"GRA15 and GRA24 single and double knockout parasites, infection of Tlr11-/- mice and human macrophages, cytokine ELISA (IL-12, IL-18, IL-1β, IFNγ), PBMC co-culture assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic knockouts (single and double), multiple host genetic backgrounds, multiple orthogonal functional readouts (in vivo and in vitro)\",\n      \"pmids\": [\"32453782\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"GRA24 promotes CCR7-mediated macrophage chemotaxis (hypermigration) via p38 MAPK signaling, enabling dissemination of T. gondii type II lineage. GRA24 and GRA15 act cooperatively: GRA15 activates NF-κB while GRA24 activates p38 MAPK, and GRA28 boosts chromatin accessibility at the Ccr7 gene locus to amplify CCR7 expression. In vivo, macrophages infected with wild-type T. gondii outcompeted GRA15/24 double-mutant-infected macrophages in migrating to secondary organs.\",\n      \"method\": \"GRA15/GRA24 single and double knockout parasites, CCR7 expression and chemotaxis assays, p38 MAPK and NF-κB signaling inhibitors, ATAC-seq for chromatin accessibility, adoptive transfer in vivo dissemination assay\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic knockouts, in vitro signaling assays, ATAC-seq, and in vivo adoptive transfer experiments; multiple orthogonal methods across two publications (peer-reviewed and preprint)\",\n      \"pmids\": [\"39207098\", \"38370679\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type II\"}]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"GRA24 is one of four nuclear-targeted effectors (TgIST, GRA16, GRA24, GRA28) necessary for a complete block of IFNγ-driven host cell death and parasite premature egress in human (but not murine) cells. GRA24's protective action in this context depends on its interaction with p38 MAPK, and this function is MYR translocon-dependent.\",\n      \"method\": \"Genome-wide loss-of-function screens in IFNγ-stimulated human cells, combinatorial GRA knockout parasites, MYR translocon inhibition, cell death and egress assays, IFNγ-driven transcription assays\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide screen followed by targeted knockouts, multiple human cell lines, functional rescue experiments, p38 MAPK interaction mechanistically confirmed\",\n      \"pmids\": [\"39292011\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"GRA24-dependent p38 MAPK signaling in host brain endothelial cells induces elevated transcription and secretion of TIMP1, which promotes non-disruptive T. gondii transmigration across polarized brain endothelial (bEnd.3) and intestinal epithelial (Caco-2) monolayers without disrupting barrier integrity. Pharmacological inhibition of p38 and use of GRA24 knockout parasites abolished TIMP1 induction.\",\n      \"method\": \"GRA24 knockout T. gondii, pharmacological p38 MAPK inhibition, transwell transmigration assays, TIMP1 ELISA and transcription analysis, barrier integrity measurements (TEER), recombinant TIMP1 supplementation\",\n      \"journal\": \"mSphere\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout and pharmacological inhibition with functional barrier transmigration readout, single lab, novel function identified\",\n      \"pmids\": [\"40265926\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"GRA24 is one of eight MYR1-dependent dense granule effectors (IST, NSM, HCE1/TEEGR, GRA16, GRA18, GRA24, GRA28, GRA84) that function cooperatively to activate the non-canonical NF-κB pathway (RelB/p52) in host cells. This activation involves parasite-driven depletion of TRAF3, stabilization of NF-κB-inducing kinase (NIK), phosphorylation and processing of p100 to p52. No single effector alone accounts for this phenotype.\",\n      \"method\": \"Combinatorial GRA knockout parasites (panel-based), immunoblotting for RelB/p52/p100/NIK/TRAF3, nuclear fractionation, genome-wide loss-of-function screening\",\n      \"journal\": \"bioRxiv : the preprint server for biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — combinatorial knockout panel with mechanistic pathway markers, preprint not yet peer-reviewed, single lab\",\n      \"pmids\": [\"41959509\"],\n      \"is_preprint\": true,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I; type II\"}]\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Deletion of GRA24 in uracil auxotrophic T. gondii strains severely abrogated antitumor immune responses, demonstrating that GRA24's trafficking to the host cell nucleus is functionally required for activating host immunity (CD8α+ dendritic cells, IL-12/IFNγ Th1 axis) that controls tumor growth.\",\n      \"method\": \"Targeted deletion of GRA24 in Δku80 vaccine strain, mouse tumor model, immune cell profiling by flow cytometry, cytokine measurements\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean gene deletion with defined in vivo phenotypic readout, single lab, functional role confirmed\",\n      \"pmids\": [\"27447180\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I Δku80\"}]\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"GRA24-induced IL-12 production acts as a virulence factor in T cell-deficient (Tcrb-/-) and IL-10-deficient mouse hosts by driving fatal cytokine storm; resistance was restored in Tcrb-/- mice by monoclonal depletion of IL-12p40, demonstrating that GRA24-driven IL-12 underlies immunopathology in the absence of regulatory T cell responses.\",\n      \"method\": \"GRA24 knockout uracil auxotroph parasites, Tcrb-/- and IL-10-/- mice, antibody depletion of IL-12p40, survival assays, cytokine profiling\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic knockout with multiple mouse genetic backgrounds and antibody depletion rescue, single lab, multiple functional readouts\",\n      \"pmids\": [\"39440975\"],\n      \"is_preprint\": false,\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I uracil auxotroph (OMP)\"}]\n    }\n  ],\n  \"current_model\": \"GRA24 is a Toxoplasma gondii dense granule protein secreted into the parasitophorous vacuole that is exported beyond the vacuole membrane via the MYR1/MYR2/MYR3 translocon complex (facilitated by TgASP5) and traffics to the host cell nucleus, where the intrinsically disordered protein binds two p38α MAPK molecules via an adapted kinase interaction motif (KIM) to bypass canonical MAPK activation, driving sustained autophosphorylation and nuclear translocation of p38α, upregulation of Egr-1 and c-Fos, production of proinflammatory cytokines (IL-12, IL-1β, MCP-1), CCR7-mediated macrophage hypermigration for parasite dissemination, TIMP1-mediated barrier transmigration, and suppression of IFNγ-driven host cell death; GRA24 additionally acts as a negative regulator of bradyzoite differentiation and, in concert with other GRA effectors, contributes to non-canonical NF-κB pathway activation in infected host cells.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"GRA24 is a Toxoplasma gondii dense granule effector that is secreted into the parasitophorous vacuole, exported into the host cell, and traffics to the host nucleus, where it functions as a non-canonical activator of host p38\\u03b1 MAP kinase to reprogram infected-cell signaling and drive proinflammatory responses [#0]. GRA24 is intrinsically disordered and binds two p38\\u03b1 molecules independently through an adapted kinase interaction motif (KIM), forming a stable complex that bypasses the canonical MAPK cascade; the recombinant GRA24\\u2013p38\\u03b1 complex is the sole factor required to activate p38\\u03b1 in vitro [#1]. Downstream, sustained p38\\u03b1 autophosphorylation and nuclear translocation drive Egr-1 and c-Fos upregulation and synthesis of proinflammatory cytokines including IL-12 and MCP-1 [#0], a response that is MyD88-independent and does not require active parasite proliferation [#8]. Export of GRA24 beyond the vacuole requires the Golgi aspartyl protease TgASP5 [#2] and the PVM-resident MYR1/MYR2/MYR3 translocon complex [#3, #4], and this export is stage-specific, being absent in bradyzoite cysts [#6]. Functionally, GRA24 acts together with other GRA effectors to shape host immunity: it cooperates with GRA15 (and GRA28) to drive CCR7-mediated macrophage hypermigration and parasite dissemination [#10], synergizes with GRA15 in IL-12/IL-18/IL-1\\u03b2 secretion and downstream IFN\\u03b3 control [#9], and contributes alongside TgIST, GRA16 and GRA28 to blocking IFN\\u03b3-driven host cell death via its p38 interaction [#11]. Independently of its host-directed role, GRA24 (TgBRADIN) acts within the parasite as a negative regulator of bradyzoite differentiation [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 2013,\n      \"claim\": \"Established that a parasite dense granule protein could reach and reprogram the host nucleus, defining GRA24 as an exported effector that non-canonically activates host p38\\u03b1 MAPK and its transcriptional output.\",\n      \"evidence\": \"Dense granule protein identification, nuclear trafficking imaging, GRA24\\u2013p38\\u03b1 co-immunoprecipitation and cytokine readouts in infected cells\",\n      \"pmids\": [\"24043761\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of p38\\u03b1 binding not resolved\", \"Mechanism of vacuolar-to-nuclear export not defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified GRA24 (TgBRADIN) as a parasite-intrinsic negative regulator of stage conversion, distinguishing a role in bradyzoite differentiation from its host-directed signaling function.\",\n      \"evidence\": \"Yeast three-hybrid screen against Compound 2 plus GRA24 gene disruption with bradyzoite differentiation assays\",\n      \"pmids\": [\"25789621\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism of differentiation suppression not elucidated\", \"Relationship to host p38\\u03b1 signaling role unclear\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Resolved a step in the export route by showing that the Golgi aspartyl protease TgASP5 is required for GRA24 export beyond the vacuole, and that this occurs independently of direct proteolytic processing of GRA24.\",\n      \"evidence\": \"ASP5 knockout parasites with immunofluorescence tracking of GRA24 localization\",\n      \"pmids\": [\"26270241\", \"26473595\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How ASP5 promotes export without cleaving GRA24 not defined\", \"Substrate/motif requirements on GRA24 unknown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Provided the structural and biochemical mechanism, showing GRA24 is intrinsically disordered and captures two p38\\u03b1 molecules via an adapted KIM to form the minimal, sufficient activating complex.\",\n      \"evidence\": \"Structural analysis, in vitro reconstitution and biophysical characterization of recombinant GRA24\\u2013human p38\\u03b1 complex\",\n      \"pmids\": [\"27889209\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the disordered protein bypasses upstream MAPK kinases mechanistically\", \"In vivo stoichiometry and turnover of the complex not established\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Defined MYR1 as an essential PVM translocation component for GRA24 delivery, linking effector export machinery to GRA24 nuclear trafficking.\",\n      \"evidence\": \"MYR1 deletion mutants with immunofluorescence for GRA24 nuclear localization and subcellular fractionation\",\n      \"pmids\": [\"26838724\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Composition of the full translocon not yet known\", \"Direct interaction between GRA24 and MYR1 not shown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"RH\"}]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrated the in vivo functional consequence of GRA24 export, showing its nuclear trafficking is required to activate host antitumor immunity via CD8\\u03b1+ DCs and the IL-12/IFN\\u03b3 Th1 axis.\",\n      \"evidence\": \"Targeted GRA24 deletion in \\u0394ku80 vaccine strain, mouse tumor model, immune cell profiling and cytokine measurement\",\n      \"pmids\": [\"27447180\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct causal chain from p38\\u03b1 to specific immune output not dissected\", \"Contribution relative to other effectors not quantified\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I \\u0394ku80\"}]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Extended the translocon model by identifying MYR2 and MYR3 as MYR1-associated PVM components required for GRA24 translocation, and provided evidence for a translocon-based unfolding mechanism.\",\n      \"evidence\": \"Genetic screen, MYR2/MYR3 deletion, reciprocal Co-IP/co-localization at PVM, GFP tracking, and DHFR-fusion transport-block experiments\",\n      \"pmids\": [\"29357375\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular architecture of the translocon pore not resolved\", \"Energetics/driving force of unfolding-coupled transport unknown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established stage-specificity of GRA24 export, showing it is not secreted beyond the cyst wall in bradyzoites, restricting its host-nuclear function to the tachyzoite stage.\",\n      \"evidence\": \"Tetracycline-inducible HA-tagged GRA24 expression in bradyzoite cysts with immunofluorescence localization\",\n      \"pmids\": [\"30406043\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism silencing export in bradyzoites unknown\", \"Whether translocon is downregulated or effector retained is not distinguished\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Connected GRA24\\u2013p38 signaling to dendritic cell reprogramming, showing GRA24 drives sustained Egr-1 to suppress DC maturation and modulate cytokine output independently of MyD88.\",\n      \"evidence\": \"GRA24 knockout parasites, Egr-1 siRNA, pharmacological p38 inhibition, nuclear Egr-1 imaging, cytokine and surface-marker flow cytometry in primary DCs\",\n      \"pmids\": [\"31681626\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream transcriptional targets of Egr-1 not mapped\", \"Net effect on protective vs suppressive immunity not resolved\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed that GRA24-driven late-phase p38 activation in macrophages promotes a host-protective Th1/IL-12 response independent of TLR/MyD88 and parasite replication.\",\n      \"evidence\": \"GRA24 knockout in cps1-1 strain, BMDM infection, IL-12-YFP reporter mice, p38 phosphorylation assays, cytokine profiling in MyD88-/- mice\",\n      \"pmids\": [\"32413093\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How p38 activation kinetics translate to sustained IL-12 transcription not detailed\", \"In vivo cellular source of protective cytokines not fully defined\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"cps1-1 (Type I uracil auxotroph)\"}]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Placed GRA24 within a cooperative effector program, showing synergy with GRA15 in driving IL-12/IL-18/IL-1\\u03b2 secretion and downstream IFN\\u03b3-mediated control of the parasite.\",\n      \"evidence\": \"GRA15/GRA24 single and double knockout parasites, Tlr11-/- mice and human macrophages, cytokine ELISA and PBMC co-culture\",\n      \"pmids\": [\"32453782\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular convergence of GRA15 (NF-\\u03baB) and GRA24 (p38) pathways not resolved\", \"Human-specific vs murine differences not fully explored\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a dissemination mechanism in which GRA24-driven p38, cooperating with GRA15 (NF-\\u03baB) and GRA28 (chromatin accessibility at Ccr7), promotes CCR7-mediated macrophage hypermigration to secondary organs.\",\n      \"evidence\": \"GRA15/GRA24 single and double knockouts, CCR7 and chemotaxis assays, p38/NF-\\u03baB inhibitors, ATAC-seq, in vivo adoptive transfer dissemination\",\n      \"pmids\": [\"39207098\", \"38370679\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct transcriptional target of GRA24-p38 at Ccr7 not shown\", \"Relative contribution of each effector in vivo not quantified\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type II\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified a barrier-crossing function whereby GRA24-p38 signaling in host endothelial/epithelial cells induces TIMP1 to enable non-disruptive parasite transmigration.\",\n      \"evidence\": \"GRA24 knockout parasites, p38 inhibition, transwell transmigration, TIMP1 ELISA/transcription, TEER integrity, recombinant TIMP1 rescue\",\n      \"pmids\": [\"40265926\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which TIMP1 facilitates transmigration not defined\", \"Single-lab observation of this specific function\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Positioned GRA24 among nuclear effectors required to block IFN\\u03b3-driven host cell death, with its protective action depending on p38 interaction and MYR translocon delivery in human but not murine cells.\",\n      \"evidence\": \"Genome-wide loss-of-function screens in IFN\\u03b3-stimulated human cells, combinatorial GRA knockouts, MYR inhibition, cell death and egress assays\",\n      \"pmids\": [\"39292011\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis of human vs murine difference not resolved\", \"How p38 activation suppresses IFN\\u03b3-driven death not mechanistically detailed\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a double-edged consequence of GRA24 signaling, showing GRA24-induced IL-12 acts as a virulence factor causing fatal cytokine storm in T cell- or IL-10-deficient hosts.\",\n      \"evidence\": \"GRA24 knockout uracil auxotrophs, Tcrb-/- and IL-10-/- mice, IL-12p40 antibody depletion rescue, survival and cytokine profiling\",\n      \"pmids\": [\"39440975\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Threshold determining protective vs pathogenic IL-12 not defined\", \"Applicability to wild-type immunocompetent hosts unclear\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I uracil auxotroph (OMP)\"}]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Placed GRA24 within a cooperative MYR1-dependent effector set that activates the non-canonical NF-\\u03baB (RelB/p52) pathway via TRAF3 depletion, NIK stabilization, and p100 processing, with no single effector sufficient.\",\n      \"evidence\": \"Combinatorial GRA knockout panels, immunoblotting for RelB/p52/p100/NIK/TRAF3, nuclear fractionation, genome-wide screening (preprint)\",\n      \"pmids\": [\"41959509\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific contribution of GRA24 to this pathway not isolated\", \"Preprint not yet peer-reviewed\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"type I\"}, {\"species\": \"Toxoplasma gondii\", \"strain\": \"type II\"}]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How GRA24 engages the MYR translocon for transport and how its p38\\u03b1 activation is integrated with parallel GRA-driven NF-\\u03baB signaling to yield distinct host outcomes remain open.\",\n      \"evidence\": \"No direct evidence in the available corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Direct GRA24\\u2013translocon interaction not demonstrated\", \"Convergence of p38 and NF-\\u03baB branches on shared host targets not resolved\", \"Mechanism of bradyzoite differentiation suppression unknown\"],\n      \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0019210\", \"supporting_discovery_ids\": [1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0019901\", \"supporting_discovery_ids\": [0, 1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0033655\", \"supporting_discovery_ids\": [0], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0042025\", \"supporting_discovery_ids\": [0, 1], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0038066\", \"supporting_discovery_ids\": [0, 8, 10, 12], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0032722\", \"supporting_discovery_ids\": [0, 8, 9], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]},\n      {\"term_id\": \"GO:0007249\", \"supporting_discovery_ids\": [10, 13], \"attribution\": [{\"species\": \"Toxoplasma gondii\", \"strain\": \"\"}]}\n    ],\n    \"complexes\": [\n      \"GRA24\\u2013p38\\u03b1 (MAPK14) complex (two p38\\u03b1 per GRA24 via adapted KIM)\"\n    ],\n    \"partners\": [\n      \"host p38\\u03b1 MAPK (MAPK14)\",\n      \"TgASP5 (export dependency)\",\n      \"MYR1 (translocation dependency)\",\n      \"MYR2 (translocation dependency)\",\n      \"MYR3 (translocation dependency)\",\n      \"GRA15 (functional cooperation)\",\n      \"GRA28 (functional cooperation)\"\n    ],\n    \"other_free_text\": [\n      \"Intrinsically disordered dense granule effector exported beyond the parasitophorous vacuole membrane\",\n      \"Drives sustained/non-canonical p38\\u03b1 activation independent of MyD88 and independent of active parasite proliferation\",\n      \"Downstream host transcriptional output includes Egr-1, c-Fos, IL-12, MCP-1, and TIMP1\",\n      \"Export is stage-specific: active in tachyzoites, absent in bradyzoite cysts\",\n      \"Parasite-intrinsic role (as TgBRADIN) as negative regulator of bradyzoite differentiation\",\n      \"GRA24-driven IL-12 can be host-protective or pathogenic (cytokine storm) depending on host immune context\"\n    ]\n  },\n  \"strain_conflicts\": []\n}","audit_flag":null,"strains":[{"strain":"ME49","gene_id":"TGME49_230180","name":"GRA24","product":"dense granule protein GRA24","uniprot_acc":"A0A125YG37","chromosome":"TGME49_chrVIII","start_pos":664211},{"strain":"GT1","gene_id":"TGGT1_230180","name":null,"product":"hypothetical protein","uniprot_acc":null,"chromosome":"TGGT1_chrVIII","start_pos":662231},{"strain":"RH88","gene_id":"TGRH88_074680","name":null,"product":"unspecified product","uniprot_acc":null,"chromosome":"CM023089","start_pos":5773440},{"strain":"VEG","gene_id":"TGVEG_230180","name":null,"product":"putative transmembrane protein","uniprot_acc":"B6KJB6","chromosome":"TGVEG_chrVIII","start_pos":671846}]}