{"gene":"PRKCQ","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":1993,"finding":"PKCθ (PRKCQ) was molecularly cloned and characterized as a novel serine/threonine kinase member of the PKC family, expressed predominantly in hematopoietic cells (T cells, thymocytes). It encodes an ~82 kDa protein, lacks the Ca2+-binding C2 domain (Ca2+-independent), and shows highest homology to PKCδ.","method":"cDNA cloning, PCR, RNase protection assay, immunoprecipitation with isoform-specific antiserum","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — original cloning paper with multiple orthogonal methods, foundational discovery","pmids":["8444877"],"is_preprint":false},{"year":1997,"finding":"PKCθ selectively translocates to the T cell–APC contact site (immunological synapse) upon antigen-specific T cell activation, where its kinase activity is selectively increased; other PKC isoforms (α, βI, βII, δ, ε) do not translocate to this site.","method":"Digital immunofluorescence microscopy of T cell–APC conjugates; in vitro kinase activity assay of PKC immunoprecipitates","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1–2 — direct localization with functional kinase assay, landmark paper with >400 citations","pmids":["8985252"],"is_preprint":false},{"year":1997,"finding":"PKCθ is cleaved by Caspase-3 in the V3 domain at DEVD354/K during apoptosis; overexpression of the cleaved kinase-active fragment (but not full-length or kinase-inactive PKCθ) induces apoptosis (sub-G1 DNA, nuclear fragmentation, lethality).","method":"In vitro Caspase-3 cleavage assay, overexpression of wild-type and mutant PKCθ constructs, DNA content analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — reconstituted in vitro cleavage with mutagenesis and functional readout","pmids":["9252332"],"is_preprint":false},{"year":1998,"finding":"PKCθ (but not PKCα or PKCε) cooperates with calcineurin to synergistically activate JNK and drive c-jun and IL-2 promoter transcription in T lymphocytes; this cooperation converges on or upstream of Rac.","method":"Isoform-selective PKC downregulation, pharmacological inhibition (cyclosporin A), reporter gene assays, dominant-negative constructs in Jurkat T cells","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — epistasis via genetic and pharmacological tools, replicated with multiple approaches","pmids":["9606192"],"is_preprint":false},{"year":1998,"finding":"The human PRKCQ gene locus maps to chromosome 10p15, spans ~62 kb, and is composed of 15 coding exons and 14 introns; 7 of 14 intron positions are conserved with the Drosophila dPRKC gene.","method":"FISH, P1 genomic clone isolation, long-range PCR, DNA sequencing","journal":"Molecular & general genetics : MGG","confidence":"High","confidence_rationale":"Tier 1 — direct genomic characterization by multiple complementary methods","pmids":["9790596"],"is_preprint":false},{"year":2000,"finding":"PKCθ is required for TCR-mediated NF-κB activation in mature T lymphocytes but is dispensable for TCR-dependent thymocyte development; TNF-α- and IL-1-induced NF-κB activation is unaffected in PKCθ-/- cells; TCR-induced JNK activation is normal in PKCθ-/- T cells.","method":"PKCθ knockout mice, T cell activation assays, NF-κB reporter/EMSA, cytokine stimulation","journal":"Nature","confidence":"High","confidence_rationale":"Tier 2 — clean genetic KO with specific phenotypic readouts, >700 citations","pmids":["10746729"],"is_preprint":false},{"year":2000,"finding":"PKCθ is uniquely capable among PKC isoforms of activating NF-κB and the CD28RE/AP-1 element in T cells; CD28 costimulation enhances PKCθ membrane translocation and catalytic activation; PKCθ-mediated NF-κB activation involves IKKβ/IκB signaling cascade.","method":"Reporter gene assays, PKC isoform overexpression, pharmacological PKCθ inhibitor, IκB degradation inhibitor (MG132), T cell transfection","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — multiple isoform comparisons, pharmacological and genetic approaches","pmids":["10716728"],"is_preprint":false},{"year":2004,"finding":"PKCθ phosphorylates IRS-1 at Ser1101, blocking IRS-1 tyrosine phosphorylation and downstream Akt pathway activation; mutation of Ser1101 to alanine renders IRS-1 insensitive to PKCθ and restores insulin signaling.","method":"In vitro kinase assay, site-directed mutagenesis (S1101A), IRS-1 phosphorylation and Akt activation assays in cultured cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase assay with mutagenesis validation, mechanistically specific","pmids":["15364919"],"is_preprint":false},{"year":2004,"finding":"PD-1 engagement inhibits TCR-mediated phosphorylation of ZAP70 and its association with CD3ζ, and attenuates PKCθ activation-loop phosphorylation; the phosphorylated PD-1 ITSM motif acts as a docking site for SHP-2 and SHP-1 in vitro.","method":"T cell stimulation with cognate TCR signal + PD-1 ligation, immunoprecipitation, phosphopeptide pull-down assays","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 2 — direct biochemical measurement of PKCθ phosphorylation state with receptor-level mechanistic dissection","pmids":["15358536"],"is_preprint":false},{"year":2005,"finding":"PKCθ phosphorylates CARMA1 on serine residues within its linker region (S564, S649, S657 per one study; Ser552 per another), converting CARMA1 from an inactive to an active conformer that recruits downstream IKK signalosome components to the immunological synapse, thereby enabling TCR-induced NF-κB activation.","method":"Co-IP, in vitro kinase assay, site-directed mutagenesis of CARMA1 serine residues, reconstitution in CARMA1-deficient T cells, lipid raft fractionation","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro kinase assay plus genetic rescue in CARMA1-deficient cells, independently replicated by two groups in the same issue","pmids":["16356855","16356856"],"is_preprint":false},{"year":2010,"finding":"PKCθ is recruited to the immunological synapse of effector T cells but is sequestered away from the regulatory T cell (Treg) immunological synapse; PKCθ blockade enhances Treg suppressive function, and inhibition of PKCθ protects Tregs from TNF-α-mediated inactivation, including restoration of defective Tregs from rheumatoid arthritis patients.","method":"Immunofluorescence microscopy, pharmacological PKCθ inhibitor, Treg suppression assays in vitro and in vivo (colitis model)","journal":"Science (New York, N.Y.)","confidence":"High","confidence_rationale":"Tier 2 — direct localization experiment linked to functional consequence, multiple in vitro and in vivo readouts","pmids":["20339032"],"is_preprint":false},{"year":2011,"finding":"PRKCQ is a direct transcriptional target of the hematopoietic transcription factor RUNX1 in megakaryocytic cells; RUNX1 binds the PRKCQ promoter at a consensus site (ACCGCA at −1088 to −1069 bp) and positively regulates PKCθ expression; RUNX1 mutation/haplodeficiency reduces PKCθ protein, contributing to impaired platelet function.","method":"Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), RUNX1 overexpression and siRNA knockdown with promoter-reporter assays","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"High","confidence_rationale":"Tier 1–2 — ChIP + EMSA + promoter mutagenesis + loss-of-function with multiple orthogonal methods","pmids":["21252065"],"is_preprint":false},{"year":2014,"finding":"Acute lipid infusion in humans transiently increases cytosolic diacylglycerol (DAG) content, which is temporally associated with PKCθ activation, increased IRS-1 Ser1101 phosphorylation, and inhibition of insulin-stimulated IRS-1 tyrosine phosphorylation and AKT2 phosphorylation, supporting DAG-activated PKCθ as a key mechanism of lipid-induced muscle insulin resistance.","method":"Serial muscle biopsies during lipid infusion, DAG and PKCθ activation measurements, IRS-1 and AKT phosphorylation assays; compared in lean, obese, and type 2 diabetic subjects","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — direct human biochemical measurements with temporal association across multiple subject groups","pmids":["24979806"],"is_preprint":false},{"year":2016,"finding":"PRKCQ/PKCθ promotes oncogenic phenotypes in breast epithelial cells: kinase-active (but not kinase-inactive) PKCθ drives Erk/MAPK-dependent Rb phosphorylation and cell-cycle progression under growth-factor-deprived conditions, as well as anchorage-independent survival and migration. Downregulation of PRKCQ in TNBC cells enhances anoikis and impairs xenograft tumor growth.","method":"Gain- and loss-of-function (kinase-active/inactive cDNA, shRNA), kinase inhibitor (AEB071), Erk/MAPK and Rb phosphorylation assays, 3D culture, xenograft tumor models","journal":"Breast cancer research : BCR","confidence":"High","confidence_rationale":"Tier 2 — kinase-inactive mutant controls with multiple readouts in vitro and in vivo","pmids":["27663795"],"is_preprint":false},{"year":2020,"finding":"PRKCQ/PKCθ regulates chemotherapy sensitivity in TNBC cells via kinase-activity-dependent suppression of pro-apoptotic Bim; kinase-active (but not kinase-inactive) PKCθ overexpression suppresses Bim, and PRKCQ inhibition restores Bim expression and enhances apoptosis induced by paclitaxel or doxorubicin.","method":"shRNA/cDNA modulation, kinase-inactive mutant, small-molecule PKCθ inhibitor (17k), Bim/Bcl2 family western blot, apoptosis assays","journal":"Breast cancer research : BCR","confidence":"High","confidence_rationale":"Tier 2 — kinase-inactive mutant controls, pharmacological inhibitor phenocopying, Bim rescue experiment","pmids":["32600444"],"is_preprint":false},{"year":2021,"finding":"Prkcq in Schwann cells regulates their proliferation, migration, and apoptosis after sciatic nerve injury, acting through the β-catenin, c-fos, and p-c-jun/c-jun pathways; Prkcq expression decreases during nerve repair, and modulation of its levels alters Wallerian degeneration and regeneration.","method":"In vivo rat sciatic nerve injury model, siRNA/overexpression in Schwann cells, pathway analysis (β-catenin, c-fos, c-jun phosphorylation), proliferation and apoptosis assays","journal":"Experimental neurology","confidence":"Medium","confidence_rationale":"Tier 3 — single lab, pathway placements inferred from expression changes and cell phenotypes without direct kinase-substrate assay","pmids":["34418453"],"is_preprint":false},{"year":2023,"finding":"miR-128-1-5p directly targets the 3'-UTR of PRKCQ mRNA (validated by luciferase assay) to suppress PRKCQ protein, thereby inhibiting colorectal cancer cell proliferation and inducing apoptosis.","method":"Luciferase reporter assay, western blot, CCK-8, colony formation, TUNEL, subcutaneous tumor model","journal":"Cancer biology & therapy","confidence":"Medium","confidence_rationale":"Tier 3 — single lab, luciferase validation of miRNA-PRKCQ interaction with functional phenotype readout","pmids":["37358216"],"is_preprint":false},{"year":2025,"finding":"PRKCQ is dispensable for spermatogenesis and male fertility in mice; Prkcq-/- males generated by CRISPR/Cas9 show no defects in spermatogenic cells, sperm parameters, or sperm motility despite PRKCQ being highly expressed in testis.","method":"CRISPR/Cas9 knockout mice, histology, immunofluorescence, computer-assisted sperm analysis, qPCR","journal":"Cell biology international","confidence":"High","confidence_rationale":"Tier 2 — clean KO with comprehensive phenotypic analysis using multiple orthogonal methods","pmids":["40051302"],"is_preprint":false},{"year":2025,"finding":"Prkcq in ILC2 cells regulates expression of IL-4, IL-5, and IL-13; Prkcq knockdown in ILC2s suppresses these cytokines and reduces tissue-resident macrophage (TRM) abundance in chronic pancreatitis, ultimately alleviating pancreatic fibrosis in a mouse model.","method":"Single-cell sequencing data analysis, DBTC-induced CP mouse model, Prkcq knockdown in ILC2s, H&E/Masson/Sirius Red staining, ILC2 and TRM quantification","journal":"Archives of biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 3 — single lab, knockdown with functional readout but mechanistic link between PRKCQ and cytokine transcription not directly established","pmids":["40706949"],"is_preprint":false},{"year":2026,"finding":"Ganoderma lucidum triterpenoids (GLTs) inhibit adipogenesis via upregulation of PRKCQ in adipocytes; deletion of PRKCQ reverses the anti-adipogenic effect of GLT, identifying PRKCQ as a mediator of the anti-obesity activity of GLTs.","method":"High-fat diet mouse model, preadipocyte differentiation assays, Prkcq deletion, adipogenic gene expression (PPARγ, C/EBPα, FASN, SCD-1), network pharmacology","journal":"Foods (Basel, Switzerland)","confidence":"Medium","confidence_rationale":"Tier 3 — single lab, genetic deletion with functional readout but downstream mechanism of PRKCQ action in adipogenesis not resolved","pmids":["41596924"],"is_preprint":false}],"current_model":"PRKCQ (PKCθ) is a Ca2+-independent serine/threonine kinase that selectively translocates to the immunological synapse upon T cell activation, where it is uniquely required for TCR/CD28-induced NF-κB activation by phosphorylating CARMA1 (converting it to an active conformer that recruits the IKK signalosome); it also cooperates with calcineurin to activate JNK and IL-2 transcription, is cleaved and activated by Caspase-3 during apoptosis, and phosphorylates IRS-1 at Ser1101 to inhibit insulin signaling downstream of diacylglycerol—a mechanism central to lipid-induced muscle insulin resistance—while its synapse localization in Treg cells is suppressed, providing a negative feedback on regulatory T cell function."},"narrative":{"teleology":[{"year":1993,"claim":"Molecular cloning established PRKCQ as a novel, calcium-independent PKC isoform with predominant hematopoietic expression, positioning it as a candidate signaling kinase in T cells.","evidence":"cDNA cloning, RNase protection assay, and immunoprecipitation with isoform-specific antisera","pmids":["8444877"],"confidence":"High","gaps":["No substrate or signaling pathway yet assigned","Enzymatic regulation by DAG/lipid cofactors not characterized at this stage"]},{"year":1997,"claim":"Discovery that PKCθ selectively translocates to the T cell–APC contact site, while other PKC isoforms do not, established its unique spatial role at the immunological synapse and suggested a non-redundant function in antigen recognition.","evidence":"Digital immunofluorescence microscopy of T cell–APC conjugates with isoform-specific antibodies and in vitro kinase assays","pmids":["8985252"],"confidence":"High","gaps":["Mechanism of selective synapse recruitment unknown","Downstream substrates at the synapse not identified"]},{"year":1997,"claim":"Identification of caspase-3 cleavage at DEVD354 in the V3 domain revealed a mechanism by which apoptotic signaling converts PKCθ into a constitutively active pro-apoptotic fragment, linking PKCθ to programmed cell death.","evidence":"In vitro caspase-3 cleavage assay, overexpression of wild-type versus kinase-inactive truncation mutants, DNA content analysis","pmids":["9252332"],"confidence":"High","gaps":["Physiological relevance of caspase-cleaved fragment in primary T cells not tested","Whether cleavage acts as positive feedback during T cell apoptosis unknown"]},{"year":1998,"claim":"Demonstration that PKCθ cooperates with calcineurin to synergistically activate JNK and IL-2 transcription, converging on or upstream of Rac, identified the first signaling pathway through which PKCθ drives T cell effector gene expression.","evidence":"Isoform-selective downregulation, cyclosporin A, dominant-negative constructs, and reporter assays in Jurkat T cells","pmids":["9606192"],"confidence":"High","gaps":["Direct kinase substrate in the calcineurin–PKCθ–JNK axis not identified","In vivo relevance not addressed"]},{"year":2000,"claim":"PKCθ knockout mice demonstrated that PKCθ is essential and non-redundant for TCR-mediated NF-κB activation in mature T cells but dispensable for thymocyte development, establishing NF-κB as the critical PKCθ-dependent pathway; CD28 costimulation was shown to specifically enhance PKCθ membrane translocation and catalytic activation.","evidence":"Prkcq−/− mice with NF-κB EMSA/reporter, cytokine stimulation controls; PKC isoform overexpression and inhibitor studies in parallel","pmids":["10746729","10716728"],"confidence":"High","gaps":["Direct phosphorylation substrate linking PKCθ to IKK not yet identified","Whether PKCθ acts through a scaffold protein unknown"]},{"year":2004,"claim":"Identification of IRS-1 Ser1101 as a direct PKCθ phosphorylation site that blocks insulin-stimulated tyrosine phosphorylation and Akt activation provided the first mechanistic link between PKCθ and metabolic insulin resistance, extending PKCθ function beyond immunity.","evidence":"In vitro kinase assay with S1101A mutagenesis and IRS-1/Akt phosphorylation readouts in cultured cells","pmids":["15364919"],"confidence":"High","gaps":["In vivo relevance in muscle tissue not yet demonstrated at this point","Upstream DAG-PKCθ activation mechanism in metabolic context not resolved"]},{"year":2005,"claim":"The discovery that PKCθ directly phosphorylates CARMA1 on linker-region serines (S552/S564/S649/S657) to convert it to an active conformer recruiting the IKK signalosome closed the mechanistic gap between PKCθ synapse translocation and NF-κB activation.","evidence":"In vitro kinase assay, site-directed mutagenesis, reconstitution in CARMA1-deficient T cells, lipid raft fractionation; independently confirmed by two groups","pmids":["16356855","16356856"],"confidence":"High","gaps":["Structural basis for CARMA1 conformational change upon phosphorylation unknown","Whether additional PKCθ substrates at the synapse contribute to NF-κB remains open"]},{"year":2010,"claim":"The finding that PKCθ is excluded from the Treg immunological synapse and that PKCθ inhibition enhances Treg suppressive function—including rescue of dysfunctional Tregs from rheumatoid arthritis patients—revealed an unexpected dichotomy in PKCθ function between effector and regulatory T cells.","evidence":"Immunofluorescence, pharmacological PKCθ inhibitor, Treg suppression assays in vitro and in a colitis model in vivo","pmids":["20339032"],"confidence":"High","gaps":["Mechanism of PKCθ exclusion from Treg synapse not identified","Whether Treg-specific PKCθ inhibition is therapeutically feasible without compromising effector T cell immunity unclear"]},{"year":2014,"claim":"Human muscle biopsy studies during lipid infusion directly demonstrated temporal association between DAG accumulation, PKCθ activation, IRS-1 Ser1101 phosphorylation, and impaired insulin signaling, validating the DAG–PKCθ–IRS-1 axis as a central mechanism of lipid-induced insulin resistance in humans.","evidence":"Serial muscle biopsies during lipid infusion in lean, obese, and T2D subjects with DAG, PKCθ, and IRS-1/AKT phosphorylation measurements","pmids":["24979806"],"confidence":"High","gaps":["Whether genetic variation at PRKCQ locus affects insulin sensitivity unknown","Whether PKCθ inhibition reverses established insulin resistance in humans not tested"]},{"year":2016,"claim":"Extending PKCθ biology to cancer, kinase-active PKCθ was shown to drive Erk/MAPK-dependent Rb phosphorylation, anchorage-independent survival, and xenograft tumor growth in triple-negative breast cancer, with subsequent work demonstrating kinase-dependent suppression of pro-apoptotic Bim as a chemoresistance mechanism.","evidence":"Gain/loss-of-function with kinase-active versus kinase-inactive mutants, shRNA, small-molecule inhibitors, xenograft models, Bim rescue experiments","pmids":["27663795","32600444"],"confidence":"High","gaps":["Direct PKCθ substrate responsible for Bim suppression not identified","Clinical relevance of PKCθ inhibition in TNBC not established"]},{"year":null,"claim":"Key unresolved questions include the structural basis for PKCθ's unique immunological synapse recruitment, the identity of additional synapse substrates beyond CARMA1, the precise mechanism by which PKCθ is excluded from the Treg synapse, and whether selective PKCθ inhibition can achieve therapeutic separation between Treg enhancement and effector T cell suppression.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal structure of full-length PKCθ in a signaling complex","No in vivo demonstration of therapeutic PKCθ inhibitor selectivity in autoimmune disease","Mechanism of synapse-specific PKCθ exclusion in Tregs unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,7,9,13,14]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,7,9]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[1,6,10]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[12]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,3,5,6,9,10]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,5,6,7,9,12]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[2,14]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[7,12]}],"complexes":[],"partners":["CARD11","IRS1","CASP3","PPP3CA","PDCD1"],"other_free_text":[]},"mechanistic_narrative":"PRKCQ (PKCθ) is a calcium-independent, diacylglycerol-activated serine/threonine kinase of the novel PKC subfamily that functions as a central signaling node in T cell activation and metabolic regulation. Upon antigen receptor engagement and CD28 costimulation, PKCθ selectively translocates to the immunological synapse—unique among PKC isoforms—where it phosphorylates CARMA1 on linker-region serines to convert it to an active conformer that recruits the IKK signalosome, thereby serving as the essential link between TCR signaling and NF-κB activation [PMID:8985252, PMID:10746729, PMID:16356855]. PKCθ cooperates with calcineurin to activate JNK and IL-2 transcription, is sequestered from the Treg immunological synapse such that its inhibition enhances regulatory T cell suppressive function, and is cleaved by caspase-3 during apoptosis to generate a pro-apoptotic kinase fragment [PMID:9606192, PMID:20339032, PMID:9252332]. Outside the immune system, PKCθ phosphorylates IRS-1 at Ser1101 downstream of diacylglycerol accumulation, directly inhibiting insulin receptor signaling and constituting a key mechanism of lipid-induced skeletal muscle insulin resistance in humans [PMID:15364919, PMID:24979806]."},"prefetch_data":{"uniprot":{"accession":"Q04759","full_name":"Protein kinase C theta type","aliases":["nPKC-theta"],"length_aa":706,"mass_kda":81.9,"function":"Calcium-independent, phospholipid- and diacylglycerol (DAG)-dependent serine/threonine-protein kinase that mediates non-redundant functions in T-cell receptor (TCR) signaling, including T-cells activation, proliferation, differentiation and survival, by mediating activation of multiple transcription factors such as NF-kappa-B, JUN, NFATC1 and NFATC2. In TCR-CD3/CD28-co-stimulated T-cells, is required for the activation of NF-kappa-B and JUN, which in turn are essential for IL2 production, and participates in the calcium-dependent NFATC1 and NFATC2 transactivation (PubMed:21964608). Mediates the activation of the canonical NF-kappa-B pathway (NFKB1) by direct phosphorylation of CARD11 on several serine residues, inducing CARD11 association with lipid rafts and recruitment of the BCL10-MALT1 complex, which then activates IKK complex, resulting in nuclear translocation and activation of NFKB1. May also play an indirect role in activation of the non-canonical NF-kappa-B (NFKB2) pathway. In the signaling pathway leading to JUN activation, acts by phosphorylating the mediator STK39/SPAK and may not act through MAP kinases signaling. Plays a critical role in TCR/CD28-induced NFATC1 and NFATC2 transactivation by participating in the regulation of reduced inositol 1,4,5-trisphosphate generation and intracellular calcium mobilization. After costimulation of T-cells through CD28 can phosphorylate CBLB and is required for the ubiquitination and subsequent degradation of CBLB, which is a prerequisite for the activation of TCR. During T-cells differentiation, plays an important role in the development of T-helper 2 (Th2) cells following immune and inflammatory responses, and, in the development of inflammatory autoimmune diseases, is necessary for the activation of IL17-producing Th17 cells. May play a minor role in Th1 response. Upon TCR stimulation, mediates T-cell protective survival signal by phosphorylating BAD, thus protecting T-cells from BAD-induced apoptosis, and by up-regulating BCL-X(L)/BCL2L1 levels through NF-kappa-B and JUN pathways. In platelets, regulates signal transduction downstream of the ITGA2B, CD36/GP4, F2R/PAR1 and F2RL3/PAR4 receptors, playing a positive role in 'outside-in' signaling and granule secretion signal transduction. May relay signals from the activated ITGA2B receptor by regulating the uncoupling of WASP and WIPF1, thereby permitting the regulation of actin filament nucleation and branching activity of the Arp2/3 complex. May mediate inhibitory effects of free fatty acids on insulin signaling by phosphorylating IRS1, which in turn blocks IRS1 tyrosine phosphorylation and downstream activation of the PI3K/AKT pathway. Phosphorylates MSN (moesin) in the presence of phosphatidylglycerol or phosphatidylinositol. Phosphorylates PDPK1 at 'Ser-504' and 'Ser-532' and negatively regulates its ability to phosphorylate PKB/AKT1. Phosphorylates CCDC88A/GIV and inhibits its guanine nucleotide exchange factor activity (PubMed:23509302). Phosphorylates and activates LRRK1, which phosphorylates RAB proteins involved in intracellular trafficking (PubMed:36040231)","subcellular_location":"Cytoplasm; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q04759/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/PRKCQ","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"FKBP5","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/PRKCQ","total_profiled":1310},"omim":[{"mim_id":"612754","title":"GLUTAREDOXIN 3; GLRX3","url":"https://www.omim.org/entry/612754"},{"mim_id":"610859","title":"CAPPING PROTEIN REGULATOR AND MYOSIN 1 LINKER 2; CARMIL2","url":"https://www.omim.org/entry/610859"},{"mim_id":"609022","title":"RAPAMYCIN-INSENSITIVE COMPANION OF MTOR; RICTOR","url":"https://www.omim.org/entry/609022"},{"mim_id":"607210","title":"CASPASE RECRUITMENT DOMAIN-CONTAINING PROTEIN 11; CARD11","url":"https://www.omim.org/entry/607210"},{"mim_id":"606883","title":"INTERLEUKIN 1 RECEPTOR-ASSOCIATED KINASE 4; IRAK4","url":"https://www.omim.org/entry/606883"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Centriolar satellite","reliability":"Enhanced"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":30.8},{"tissue":"skeletal muscle","ntpm":115.2},{"tissue":"tongue","ntpm":104.8}],"url":"https://www.proteinatlas.org/search/PRKCQ"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q04759","domains":[{"cath_id":"2.60.40.150","chopping":"5-123","consensus_level":"high","plddt":80.9074,"start":5,"end":123},{"cath_id":"3.30.60.20","chopping":"155-286","consensus_level":"high","plddt":79.9101,"start":155,"end":286},{"cath_id":"3.30.200.20","chopping":"379-463_671-701","consensus_level":"high","plddt":87.9837,"start":379,"end":701},{"cath_id":"1.10.510.10","chopping":"468-645","consensus_level":"high","plddt":94.4887,"start":468,"end":645}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q04759","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q04759-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q04759-F1-predicted_aligned_error_v6.png","plddt_mean":79.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=PRKCQ","jax_strain_url":"https://www.jax.org/strain/search?query=PRKCQ"},"sequence":{"accession":"Q04759","fasta_url":"https://rest.uniprot.org/uniprotkb/Q04759.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q04759/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q04759"}},"corpus_meta":[{"pmid":"27663795","id":"PMC_27663795","title":"PRKCQ promotes oncogenic growth and anoikis resistance of a subset of triple-negative breast cancer cells.","date":"2016","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/27663795","citation_count":39,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32619070","id":"PMC_32619070","title":"Long noncoding RNA PRKCQ-AS1 promotes CRC cell proliferation and migration via modulating miR-1287-5p/YBX1 axis.","date":"2020","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/32619070","citation_count":29,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"32600444","id":"PMC_32600444","title":"PRKCQ inhibition enhances chemosensitivity of triple-negative breast cancer by regulating Bim.","date":"2020","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/32600444","citation_count":28,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"35447530","id":"PMC_35447530","title":"Crocin attenuates NF-κB-mediated inflammation and proliferation in breast cancer cells by down-regulating PRKCQ.","date":"2022","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/35447530","citation_count":26,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"21252065","id":"PMC_21252065","title":"Platelet protein kinase C-theta deficiency with human RUNX1 mutation: PRKCQ is a transcriptional target of RUNX1.","date":"2011","source":"Arteriosclerosis, thrombosis, and vascular biology","url":"https://pubmed.ncbi.nlm.nih.gov/21252065","citation_count":23,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"36732923","id":"PMC_36732923","title":"LncRNA PRKCQ-AS1 regulates paclitaxel resistance in triple-negative breast cancer cells through miR-361-5p/PIK3C3 mediated autophagy.","date":"2023","source":"Clinical and experimental pharmacology & physiology","url":"https://pubmed.ncbi.nlm.nih.gov/36732923","citation_count":20,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"26784953","id":"PMC_26784953","title":"Genetic Variation in the REL Gene Increases Risk of Behcet's Disease in a Chinese Han Population but That of PRKCQ Does Not.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26784953","citation_count":11,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"37358216","id":"PMC_37358216","title":"MiR-128-1-5p inhibits cell proliferation and induces cell apoptosis via targeting PRKCQ in colorectal cancer.","date":"2023","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/37358216","citation_count":11,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38092752","id":"PMC_38092752","title":"Fasting regulates mitochondrial function through lncRNA PRKCQ-AS1-mediated IGF2BPs in papillary thyroid carcinoma.","date":"2023","source":"Cell death & 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RUNX1 overexpression enhanced PKCθ protein and promoter activity, while mutation of the RUNX1 site or siRNA knockdown of RUNX1 decreased PRKCQ promoter activity and PKCθ protein levels.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), EMSA, RUNX1 overexpression/siRNA knockdown with promoter-reporter assays\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods (ChIP, EMSA, promoter mutagenesis, siRNA) in a single study\",\n      \"pmids\": [\"21252065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PKCθ kinase activity promotes Rb phosphorylation and cell-cycle progression via stimulation of Erk/MAPK signaling; kinase-inactive PKCθ overexpression fails to activate Erk/MAPK or Rb phosphorylation or promote growth-factor-independent proliferation, establishing that the catalytic activity of PRKCQ is required for these downstream effects.\",\n      \"method\": \"Gain-of-function with kinase-active vs. kinase-inactive PKCθ constructs in MCF-10A cells; western blot for pRb and pErk; proliferation assays\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean loss/gain of function with defined cellular phenotype and pathway placement in single lab\",\n      \"pmids\": [\"27663795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PKCθ (PRKCQ) promotes anoikis resistance, anchorage-independent growth, and migration in breast epithelial cells; shRNA-mediated downregulation in TNBC cells enhances anoikis and impairs 3D growth and xenograft tumor growth in vivo.\",\n      \"method\": \"shRNA knockdown, cDNA overexpression, 3D Matrigel cultures, xenograft mouse model, small-molecule kinase inhibitor (AEB071)\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss and gain of function with in vitro and in vivo phenotypic readouts in single lab\",\n      \"pmids\": [\"27663795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"PRKCQ kinase activity suppresses pro-apoptotic Bim levels in TNBC cells; kinase-inactive PRKCQ fails to suppress Bim or chemotherapy-induced apoptosis, placing PRKCQ upstream of Bim in the apoptotic pathway and demonstrating that catalytic activity is required for this regulation.\",\n      \"method\": \"shRNA/cDNA modulation of PRKCQ, kinase-inactive mutant overexpression, Bcl-2 family member western blotting, small-molecule kinase inhibitor (compound 17k), apoptosis assays\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — kinase-dead mutant plus inhibitor plus KD with specific molecular readout (Bim) in single lab\",\n      \"pmids\": [\"32600444\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Prkcq in Schwann cells regulates their proliferation, migration, and apoptosis following sciatic nerve injury through activation of the β-catenin, c-fos, and p-c-jun/c-jun signaling pathways; upregulation and downregulation of Prkcq modulated these pathways and associated cellular behaviors in vitro and in vivo.\",\n      \"method\": \"In vivo rat sciatic nerve injury model; Prkcq overexpression/knockdown in Schwann cells; western blot for β-catenin, c-fos, p-c-jun; proliferation and apoptosis assays\",\n      \"journal\": \"Experimental neurology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain/loss of function with multiple pathway readouts in single lab\",\n      \"pmids\": [\"34418453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The human PRKCQ gene locus was mapped to chromosome 10p15 by FISH analysis; the gene spans ~62 kb and is composed of 15 coding exons and 14 introns, with high conservation of intron positions compared to the Drosophila dPRKC gene.\",\n      \"method\": \"Genomic cloning from P1 libraries, FISH analysis, long-range PCR, DNA sequencing\",\n      \"journal\": \"Molecular & general genetics : MGG\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct genomic characterization with FISH mapping and sequence analysis\",\n      \"pmids\": [\"9790596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PRKCQ knockdown in ILC2 cells suppresses expression of IL-4, IL-5, and IL-13, and reduces tissue-resident macrophage (TRM) abundance in a mouse model of chronic pancreatitis, thereby alleviating pancreatic fibrosis; this places Prkcq upstream of cytokine production in ILC2s and the resulting macrophage-driven fibrosis.\",\n      \"method\": \"Prkcq knockdown in ILC2 cells (mouse model), single-cell sequencing, histological staining (H&E, Masson, Sirius Red), cytokine expression analysis\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single knockdown approach with phenotypic readout but limited mechanistic detail\",\n      \"pmids\": [\"40706949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"PRKCQ is required for the anti-adipogenic effect of Ganoderma lucidum triterpenoids (GLT); GLT upregulates PRKCQ expression in adipocytes, and PRKCQ deletion reverses the anti-adipogenic effect of GLT, indicating PRKCQ functions downstream of GLT to suppress preadipocyte differentiation and lipid accumulation.\",\n      \"method\": \"PRKCQ knockout/knockdown in adipocyte cell model, HFD mouse model, network pharmacology, adipogenic gene expression (PPARγ, C/EBPα, FASN, SCD-1)\",\n      \"journal\": \"Foods (Basel, Switzerland)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, deletion phenotype rescue approach with limited mechanistic pathway resolution\",\n      \"pmids\": [\"41596924\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PRKCQ (PKCθ) is a serine/threonine kinase whose expression in T lymphocytes, megakaryocytes, and breast epithelia is transcriptionally controlled by RUNX1; its catalytic activity drives Erk/MAPK signaling to promote Rb phosphorylation and cell-cycle progression, suppresses pro-apoptotic Bim to confer anoikis resistance, and in Schwann cells activates β-catenin/c-fos/c-jun pathways to regulate proliferation and migration during nerve repair.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"PKCθ (PRKCQ) was molecularly cloned and characterized as a novel serine/threonine kinase member of the PKC family, expressed predominantly in hematopoietic cells (T cells, thymocytes). It encodes an ~82 kDa protein, lacks the Ca2+-binding C2 domain (Ca2+-independent), and shows highest homology to PKCδ.\",\n      \"method\": \"cDNA cloning, PCR, RNase protection assay, immunoprecipitation with isoform-specific antiserum\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — original cloning paper with multiple orthogonal methods, foundational discovery\",\n      \"pmids\": [\"8444877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"PKCθ selectively translocates to the T cell–APC contact site (immunological synapse) upon antigen-specific T cell activation, where its kinase activity is selectively increased; other PKC isoforms (α, βI, βII, δ, ε) do not translocate to this site.\",\n      \"method\": \"Digital immunofluorescence microscopy of T cell–APC conjugates; in vitro kinase activity assay of PKC immunoprecipitates\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — direct localization with functional kinase assay, landmark paper with >400 citations\",\n      \"pmids\": [\"8985252\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"PKCθ is cleaved by Caspase-3 in the V3 domain at DEVD354/K during apoptosis; overexpression of the cleaved kinase-active fragment (but not full-length or kinase-inactive PKCθ) induces apoptosis (sub-G1 DNA, nuclear fragmentation, lethality).\",\n      \"method\": \"In vitro Caspase-3 cleavage assay, overexpression of wild-type and mutant PKCθ constructs, DNA content analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted in vitro cleavage with mutagenesis and functional readout\",\n      \"pmids\": [\"9252332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"PKCθ (but not PKCα or PKCε) cooperates with calcineurin to synergistically activate JNK and drive c-jun and IL-2 promoter transcription in T lymphocytes; this cooperation converges on or upstream of Rac.\",\n      \"method\": \"Isoform-selective PKC downregulation, pharmacological inhibition (cyclosporin A), reporter gene assays, dominant-negative constructs in Jurkat T cells\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — epistasis via genetic and pharmacological tools, replicated with multiple approaches\",\n      \"pmids\": [\"9606192\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The human PRKCQ gene locus maps to chromosome 10p15, spans ~62 kb, and is composed of 15 coding exons and 14 introns; 7 of 14 intron positions are conserved with the Drosophila dPRKC gene.\",\n      \"method\": \"FISH, P1 genomic clone isolation, long-range PCR, DNA sequencing\",\n      \"journal\": \"Molecular & general genetics : MGG\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct genomic characterization by multiple complementary methods\",\n      \"pmids\": [\"9790596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"PKCθ is required for TCR-mediated NF-κB activation in mature T lymphocytes but is dispensable for TCR-dependent thymocyte development; TNF-α- and IL-1-induced NF-κB activation is unaffected in PKCθ-/- cells; TCR-induced JNK activation is normal in PKCθ-/- T cells.\",\n      \"method\": \"PKCθ knockout mice, T cell activation assays, NF-κB reporter/EMSA, cytokine stimulation\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic KO with specific phenotypic readouts, >700 citations\",\n      \"pmids\": [\"10746729\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"PKCθ is uniquely capable among PKC isoforms of activating NF-κB and the CD28RE/AP-1 element in T cells; CD28 costimulation enhances PKCθ membrane translocation and catalytic activation; PKCθ-mediated NF-κB activation involves IKKβ/IκB signaling cascade.\",\n      \"method\": \"Reporter gene assays, PKC isoform overexpression, pharmacological PKCθ inhibitor, IκB degradation inhibitor (MG132), T cell transfection\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple isoform comparisons, pharmacological and genetic approaches\",\n      \"pmids\": [\"10716728\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"PKCθ phosphorylates IRS-1 at Ser1101, blocking IRS-1 tyrosine phosphorylation and downstream Akt pathway activation; mutation of Ser1101 to alanine renders IRS-1 insensitive to PKCθ and restores insulin signaling.\",\n      \"method\": \"In vitro kinase assay, site-directed mutagenesis (S1101A), IRS-1 phosphorylation and Akt activation assays in cultured cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase assay with mutagenesis validation, mechanistically specific\",\n      \"pmids\": [\"15364919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"PD-1 engagement inhibits TCR-mediated phosphorylation of ZAP70 and its association with CD3ζ, and attenuates PKCθ activation-loop phosphorylation; the phosphorylated PD-1 ITSM motif acts as a docking site for SHP-2 and SHP-1 in vitro.\",\n      \"method\": \"T cell stimulation with cognate TCR signal + PD-1 ligation, immunoprecipitation, phosphopeptide pull-down assays\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct biochemical measurement of PKCθ phosphorylation state with receptor-level mechanistic dissection\",\n      \"pmids\": [\"15358536\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"PKCθ phosphorylates CARMA1 on serine residues within its linker region (S564, S649, S657 per one study; Ser552 per another), converting CARMA1 from an inactive to an active conformer that recruits downstream IKK signalosome components to the immunological synapse, thereby enabling TCR-induced NF-κB activation.\",\n      \"method\": \"Co-IP, in vitro kinase assay, site-directed mutagenesis of CARMA1 serine residues, reconstitution in CARMA1-deficient T cells, lipid raft fractionation\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro kinase assay plus genetic rescue in CARMA1-deficient cells, independently replicated by two groups in the same issue\",\n      \"pmids\": [\"16356855\", \"16356856\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"PKCθ is recruited to the immunological synapse of effector T cells but is sequestered away from the regulatory T cell (Treg) immunological synapse; PKCθ blockade enhances Treg suppressive function, and inhibition of PKCθ protects Tregs from TNF-α-mediated inactivation, including restoration of defective Tregs from rheumatoid arthritis patients.\",\n      \"method\": \"Immunofluorescence microscopy, pharmacological PKCθ inhibitor, Treg suppression assays in vitro and in vivo (colitis model)\",\n      \"journal\": \"Science (New York, N.Y.)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment linked to functional consequence, multiple in vitro and in vivo readouts\",\n      \"pmids\": [\"20339032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"PRKCQ is a direct transcriptional target of the hematopoietic transcription factor RUNX1 in megakaryocytic cells; RUNX1 binds the PRKCQ promoter at a consensus site (ACCGCA at −1088 to −1069 bp) and positively regulates PKCθ expression; RUNX1 mutation/haplodeficiency reduces PKCθ protein, contributing to impaired platelet function.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), RUNX1 overexpression and siRNA knockdown with promoter-reporter assays\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP + EMSA + promoter mutagenesis + loss-of-function with multiple orthogonal methods\",\n      \"pmids\": [\"21252065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Acute lipid infusion in humans transiently increases cytosolic diacylglycerol (DAG) content, which is temporally associated with PKCθ activation, increased IRS-1 Ser1101 phosphorylation, and inhibition of insulin-stimulated IRS-1 tyrosine phosphorylation and AKT2 phosphorylation, supporting DAG-activated PKCθ as a key mechanism of lipid-induced muscle insulin resistance.\",\n      \"method\": \"Serial muscle biopsies during lipid infusion, DAG and PKCθ activation measurements, IRS-1 and AKT phosphorylation assays; compared in lean, obese, and type 2 diabetic subjects\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct human biochemical measurements with temporal association across multiple subject groups\",\n      \"pmids\": [\"24979806\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PRKCQ/PKCθ promotes oncogenic phenotypes in breast epithelial cells: kinase-active (but not kinase-inactive) PKCθ drives Erk/MAPK-dependent Rb phosphorylation and cell-cycle progression under growth-factor-deprived conditions, as well as anchorage-independent survival and migration. Downregulation of PRKCQ in TNBC cells enhances anoikis and impairs xenograft tumor growth.\",\n      \"method\": \"Gain- and loss-of-function (kinase-active/inactive cDNA, shRNA), kinase inhibitor (AEB071), Erk/MAPK and Rb phosphorylation assays, 3D culture, xenograft tumor models\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — kinase-inactive mutant controls with multiple readouts in vitro and in vivo\",\n      \"pmids\": [\"27663795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"PRKCQ/PKCθ regulates chemotherapy sensitivity in TNBC cells via kinase-activity-dependent suppression of pro-apoptotic Bim; kinase-active (but not kinase-inactive) PKCθ overexpression suppresses Bim, and PRKCQ inhibition restores Bim expression and enhances apoptosis induced by paclitaxel or doxorubicin.\",\n      \"method\": \"shRNA/cDNA modulation, kinase-inactive mutant, small-molecule PKCθ inhibitor (17k), Bim/Bcl2 family western blot, apoptosis assays\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — kinase-inactive mutant controls, pharmacological inhibitor phenocopying, Bim rescue experiment\",\n      \"pmids\": [\"32600444\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Prkcq in Schwann cells regulates their proliferation, migration, and apoptosis after sciatic nerve injury, acting through the β-catenin, c-fos, and p-c-jun/c-jun pathways; Prkcq expression decreases during nerve repair, and modulation of its levels alters Wallerian degeneration and regeneration.\",\n      \"method\": \"In vivo rat sciatic nerve injury model, siRNA/overexpression in Schwann cells, pathway analysis (β-catenin, c-fos, c-jun phosphorylation), proliferation and apoptosis assays\",\n      \"journal\": \"Experimental neurology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, pathway placements inferred from expression changes and cell phenotypes without direct kinase-substrate assay\",\n      \"pmids\": [\"34418453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"miR-128-1-5p directly targets the 3'-UTR of PRKCQ mRNA (validated by luciferase assay) to suppress PRKCQ protein, thereby inhibiting colorectal cancer cell proliferation and inducing apoptosis.\",\n      \"method\": \"Luciferase reporter assay, western blot, CCK-8, colony formation, TUNEL, subcutaneous tumor model\",\n      \"journal\": \"Cancer biology & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, luciferase validation of miRNA-PRKCQ interaction with functional phenotype readout\",\n      \"pmids\": [\"37358216\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PRKCQ is dispensable for spermatogenesis and male fertility in mice; Prkcq-/- males generated by CRISPR/Cas9 show no defects in spermatogenic cells, sperm parameters, or sperm motility despite PRKCQ being highly expressed in testis.\",\n      \"method\": \"CRISPR/Cas9 knockout mice, histology, immunofluorescence, computer-assisted sperm analysis, qPCR\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with comprehensive phenotypic analysis using multiple orthogonal methods\",\n      \"pmids\": [\"40051302\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Prkcq in ILC2 cells regulates expression of IL-4, IL-5, and IL-13; Prkcq knockdown in ILC2s suppresses these cytokines and reduces tissue-resident macrophage (TRM) abundance in chronic pancreatitis, ultimately alleviating pancreatic fibrosis in a mouse model.\",\n      \"method\": \"Single-cell sequencing data analysis, DBTC-induced CP mouse model, Prkcq knockdown in ILC2s, H&E/Masson/Sirius Red staining, ILC2 and TRM quantification\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, knockdown with functional readout but mechanistic link between PRKCQ and cytokine transcription not directly established\",\n      \"pmids\": [\"40706949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Ganoderma lucidum triterpenoids (GLTs) inhibit adipogenesis via upregulation of PRKCQ in adipocytes; deletion of PRKCQ reverses the anti-adipogenic effect of GLT, identifying PRKCQ as a mediator of the anti-obesity activity of GLTs.\",\n      \"method\": \"High-fat diet mouse model, preadipocyte differentiation assays, Prkcq deletion, adipogenic gene expression (PPARγ, C/EBPα, FASN, SCD-1), network pharmacology\",\n      \"journal\": \"Foods (Basel, Switzerland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, genetic deletion with functional readout but downstream mechanism of PRKCQ action in adipogenesis not resolved\",\n      \"pmids\": [\"41596924\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PRKCQ (PKCθ) is a Ca2+-independent serine/threonine kinase that selectively translocates to the immunological synapse upon T cell activation, where it is uniquely required for TCR/CD28-induced NF-κB activation by phosphorylating CARMA1 (converting it to an active conformer that recruits the IKK signalosome); it also cooperates with calcineurin to activate JNK and IL-2 transcription, is cleaved and activated by Caspase-3 during apoptosis, and phosphorylates IRS-1 at Ser1101 to inhibit insulin signaling downstream of diacylglycerol—a mechanism central to lipid-induced muscle insulin resistance—while its synapse localization in Treg cells is suppressed, providing a negative feedback on regulatory T cell function.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"PRKCQ (PKCθ) is a serine/threonine kinase that functions as a signaling hub linking extracellular cues to proliferation, survival, and immune effector programs in T lymphocytes, epithelial cells, and Schwann cells. Its catalytic activity drives Erk/MAPK signaling to phosphorylate Rb and promote cell-cycle progression, and suppresses the pro-apoptotic BH3-only protein Bim to confer anoikis resistance and chemotherapy survival in triple-negative breast cancer cells [PMID:27663795, PMID:32600444]. In Schwann cells following nerve injury, PRKCQ activates β-catenin, c-fos, and phospho-c-jun pathways to regulate proliferation, migration, and apoptosis during nerve repair [PMID:34418453]. Transcription of PRKCQ is directly controlled by RUNX1 through a consensus binding site in the PRKCQ promoter, as demonstrated in megakaryocytic cells [PMID:21252065].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Defining the genomic architecture of PRKCQ established its chromosomal location and exon–intron organization, providing the structural framework for all subsequent functional studies.\",\n      \"evidence\": \"Genomic cloning from P1 libraries, FISH mapping to chromosome 10p15, and DNA sequencing in human cells\",\n      \"pmids\": [\"9790596\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No functional or enzymatic characterization was provided\",\n        \"Tissue-specific expression patterns not systematically analyzed\"\n      ]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identifying RUNX1 as a direct transcriptional regulator of PRKCQ answered how lineage-specific expression is achieved in megakaryocytes, connecting a key hematopoietic transcription factor to PKCθ levels.\",\n      \"evidence\": \"ChIP, EMSA, promoter mutagenesis, and RUNX1 overexpression/siRNA knockdown with reporter assays in megakaryocytic cells\",\n      \"pmids\": [\"21252065\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether RUNX1 similarly controls PRKCQ transcription in T lymphocytes or other cell types was not tested\",\n        \"Other transcription factors contributing to tissue-specific PRKCQ expression remain unidentified\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Demonstrating that PRKCQ catalytic activity is required for Erk/MAPK activation, Rb phosphorylation, anoikis resistance, and anchorage-independent growth established its kinase-dependent oncogenic signaling axis in breast epithelial cells.\",\n      \"evidence\": \"Kinase-active vs. kinase-inactive PRKCQ constructs, shRNA knockdown, 3D Matrigel cultures, xenograft mouse model, and small-molecule inhibitor (AEB071) in MCF-10A and TNBC cells\",\n      \"pmids\": [\"27663795\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct PRKCQ substrates linking it to Erk activation were not identified\",\n        \"Whether these pathways operate identically in primary human tumors was not confirmed\",\n        \"Mechanism by which PRKCQ suppresses anoikis beyond Erk remains unresolved\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Placing PRKCQ upstream of Bim suppression resolved how its kinase activity protects TNBC cells from chemotherapy-induced apoptosis, identifying a specific pro-apoptotic target of the pathway.\",\n      \"evidence\": \"shRNA/cDNA modulation, kinase-inactive mutant, small-molecule inhibitor (compound 17k), Bcl-2 family western blotting, and apoptosis assays in TNBC cells\",\n      \"pmids\": [\"32600444\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether PRKCQ phosphorylates Bim directly or acts through intermediate kinases was not determined\",\n        \"The mechanism of Bim downregulation (transcriptional vs. post-translational) was not resolved\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showing that Prkcq controls Schwann cell proliferation, migration, and apoptosis via β-catenin/c-fos/c-jun pathways extended its functional repertoire beyond immune and epithelial cells to peripheral nerve repair.\",\n      \"evidence\": \"Prkcq overexpression/knockdown in Schwann cells; rat sciatic nerve injury model; western blot for β-catenin, c-fos, p-c-jun\",\n      \"pmids\": [\"34418453\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct phosphorylation targets of PKCθ in Schwann cells were not identified\",\n        \"How PKCθ activates β-catenin signaling mechanistically remains unknown\",\n        \"Whether these findings translate to human nerve injury is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Linking PRKCQ to ILC2 cytokine production and downstream macrophage-driven pancreatic fibrosis broadened its immune role beyond T cells to innate lymphoid cell function in chronic inflammation.\",\n      \"evidence\": \"Prkcq knockdown in ILC2 cells, single-cell sequencing, histological staining in mouse chronic pancreatitis model\",\n      \"pmids\": [\"40706949\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Single knockdown approach without genetic rescue or pharmacologic confirmation\",\n        \"Signaling pathway connecting PKCθ to IL-4/IL-5/IL-13 transcription in ILC2s was not delineated\",\n        \"Not independently confirmed by other laboratories\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct substrates of PKCθ that mediate Erk activation, Bim suppression, and β-catenin signaling remain unidentified, and whether its diverse cell-type-specific roles converge on shared or distinct downstream phosphorylation events is unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No unbiased phosphoproteomics identifying direct PKCθ substrates in any cell type\",\n        \"Structural basis for PKCθ substrate selectivity is not established\",\n        \"Whether PRKCQ kinase activity versus scaffolding contributes to all reported phenotypes has not been systematically dissected\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 3, 4]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 3, 4]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"RUNX1\",\n      \"BCL2L11\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"PRKCQ (PKCθ) is a calcium-independent, diacylglycerol-activated serine/threonine kinase of the novel PKC subfamily that functions as a central signaling node in T cell activation and metabolic regulation. Upon antigen receptor engagement and CD28 costimulation, PKCθ selectively translocates to the immunological synapse—unique among PKC isoforms—where it phosphorylates CARMA1 on linker-region serines to convert it to an active conformer that recruits the IKK signalosome, thereby serving as the essential link between TCR signaling and NF-κB activation [PMID:8985252, PMID:10746729, PMID:16356855]. PKCθ cooperates with calcineurin to activate JNK and IL-2 transcription, is sequestered from the Treg immunological synapse such that its inhibition enhances regulatory T cell suppressive function, and is cleaved by caspase-3 during apoptosis to generate a pro-apoptotic kinase fragment [PMID:9606192, PMID:20339032, PMID:9252332]. Outside the immune system, PKCθ phosphorylates IRS-1 at Ser1101 downstream of diacylglycerol accumulation, directly inhibiting insulin receptor signaling and constituting a key mechanism of lipid-induced skeletal muscle insulin resistance in humans [PMID:15364919, PMID:24979806].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Molecular cloning established PRKCQ as a novel, calcium-independent PKC isoform with predominant hematopoietic expression, positioning it as a candidate signaling kinase in T cells.\",\n      \"evidence\": \"cDNA cloning, RNase protection assay, and immunoprecipitation with isoform-specific antisera\",\n      \"pmids\": [\"8444877\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No substrate or signaling pathway yet assigned\", \"Enzymatic regulation by DAG/lipid cofactors not characterized at this stage\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Discovery that PKCθ selectively translocates to the T cell–APC contact site, while other PKC isoforms do not, established its unique spatial role at the immunological synapse and suggested a non-redundant function in antigen recognition.\",\n      \"evidence\": \"Digital immunofluorescence microscopy of T cell–APC conjugates with isoform-specific antibodies and in vitro kinase assays\",\n      \"pmids\": [\"8985252\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of selective synapse recruitment unknown\", \"Downstream substrates at the synapse not identified\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Identification of caspase-3 cleavage at DEVD354 in the V3 domain revealed a mechanism by which apoptotic signaling converts PKCθ into a constitutively active pro-apoptotic fragment, linking PKCθ to programmed cell death.\",\n      \"evidence\": \"In vitro caspase-3 cleavage assay, overexpression of wild-type versus kinase-inactive truncation mutants, DNA content analysis\",\n      \"pmids\": [\"9252332\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological relevance of caspase-cleaved fragment in primary T cells not tested\", \"Whether cleavage acts as positive feedback during T cell apoptosis unknown\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Demonstration that PKCθ cooperates with calcineurin to synergistically activate JNK and IL-2 transcription, converging on or upstream of Rac, identified the first signaling pathway through which PKCθ drives T cell effector gene expression.\",\n      \"evidence\": \"Isoform-selective downregulation, cyclosporin A, dominant-negative constructs, and reporter assays in Jurkat T cells\",\n      \"pmids\": [\"9606192\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct kinase substrate in the calcineurin–PKCθ–JNK axis not identified\", \"In vivo relevance not addressed\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"PKCθ knockout mice demonstrated that PKCθ is essential and non-redundant for TCR-mediated NF-κB activation in mature T cells but dispensable for thymocyte development, establishing NF-κB as the critical PKCθ-dependent pathway; CD28 costimulation was shown to specifically enhance PKCθ membrane translocation and catalytic activation.\",\n      \"evidence\": \"Prkcq−/− mice with NF-κB EMSA/reporter, cytokine stimulation controls; PKC isoform overexpression and inhibitor studies in parallel\",\n      \"pmids\": [\"10746729\", \"10716728\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct phosphorylation substrate linking PKCθ to IKK not yet identified\", \"Whether PKCθ acts through a scaffold protein unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identification of IRS-1 Ser1101 as a direct PKCθ phosphorylation site that blocks insulin-stimulated tyrosine phosphorylation and Akt activation provided the first mechanistic link between PKCθ and metabolic insulin resistance, extending PKCθ function beyond immunity.\",\n      \"evidence\": \"In vitro kinase assay with S1101A mutagenesis and IRS-1/Akt phosphorylation readouts in cultured cells\",\n      \"pmids\": [\"15364919\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance in muscle tissue not yet demonstrated at this point\", \"Upstream DAG-PKCθ activation mechanism in metabolic context not resolved\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"The discovery that PKCθ directly phosphorylates CARMA1 on linker-region serines (S552/S564/S649/S657) to convert it to an active conformer recruiting the IKK signalosome closed the mechanistic gap between PKCθ synapse translocation and NF-κB activation.\",\n      \"evidence\": \"In vitro kinase assay, site-directed mutagenesis, reconstitution in CARMA1-deficient T cells, lipid raft fractionation; independently confirmed by two groups\",\n      \"pmids\": [\"16356855\", \"16356856\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis for CARMA1 conformational change upon phosphorylation unknown\", \"Whether additional PKCθ substrates at the synapse contribute to NF-κB remains open\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"The finding that PKCθ is excluded from the Treg immunological synapse and that PKCθ inhibition enhances Treg suppressive function—including rescue of dysfunctional Tregs from rheumatoid arthritis patients—revealed an unexpected dichotomy in PKCθ function between effector and regulatory T cells.\",\n      \"evidence\": \"Immunofluorescence, pharmacological PKCθ inhibitor, Treg suppression assays in vitro and in a colitis model in vivo\",\n      \"pmids\": [\"20339032\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of PKCθ exclusion from Treg synapse not identified\", \"Whether Treg-specific PKCθ inhibition is therapeutically feasible without compromising effector T cell immunity unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Human muscle biopsy studies during lipid infusion directly demonstrated temporal association between DAG accumulation, PKCθ activation, IRS-1 Ser1101 phosphorylation, and impaired insulin signaling, validating the DAG–PKCθ–IRS-1 axis as a central mechanism of lipid-induced insulin resistance in humans.\",\n      \"evidence\": \"Serial muscle biopsies during lipid infusion in lean, obese, and T2D subjects with DAG, PKCθ, and IRS-1/AKT phosphorylation measurements\",\n      \"pmids\": [\"24979806\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether genetic variation at PRKCQ locus affects insulin sensitivity unknown\", \"Whether PKCθ inhibition reverses established insulin resistance in humans not tested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extending PKCθ biology to cancer, kinase-active PKCθ was shown to drive Erk/MAPK-dependent Rb phosphorylation, anchorage-independent survival, and xenograft tumor growth in triple-negative breast cancer, with subsequent work demonstrating kinase-dependent suppression of pro-apoptotic Bim as a chemoresistance mechanism.\",\n      \"evidence\": \"Gain/loss-of-function with kinase-active versus kinase-inactive mutants, shRNA, small-molecule inhibitors, xenograft models, Bim rescue experiments\",\n      \"pmids\": [\"27663795\", \"32600444\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct PKCθ substrate responsible for Bim suppression not identified\", \"Clinical relevance of PKCθ inhibition in TNBC not established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis for PKCθ's unique immunological synapse recruitment, the identity of additional synapse substrates beyond CARMA1, the precise mechanism by which PKCθ is excluded from the Treg synapse, and whether selective PKCθ inhibition can achieve therapeutic separation between Treg enhancement and effector T cell suppression.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal structure of full-length PKCθ in a signaling complex\", \"No in vivo demonstration of therapeutic PKCθ inhibitor selectivity in autoimmune disease\", \"Mechanism of synapse-specific PKCθ exclusion in Tregs unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 7, 9, 13, 14]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 7, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [1, 6, 10]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 3, 5, 6, 9, 10]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 5, 6, 7, 9, 12]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 14]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [7, 12]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"CARD11\",\n      \"IRS1\",\n      \"CASP3\",\n      \"PPP3CA\",\n      \"PDCD1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}