{"gene":"PCK1","run_date":"2026-04-29T11:37:58","timeline":{"discoveries":[{"year":2020,"finding":"AKT phosphorylates PCK1 at Ser90, triggering its translocation from cytosol to the endoplasmic reticulum, where phosphorylated PCK1 acts as a GTP-dependent protein kinase to phosphorylate INSIG1 at Ser207 and INSIG2 at Ser151. This phosphorylation reduces sterol binding to INSIG1/2, disrupts INSIG–SCAP interaction, and releases the SCAP–SREBP complex to translocate to the Golgi, activating lipogenic gene transcription.","method":"In vitro kinase assays, site-directed mutagenesis, Co-IP, subcellular fractionation, ER translocation imaging, xenograft tumor models, patient HCC sample correlation","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution of kinase activity, mutagenesis of phosphosites, multiple orthogonal methods in single study, replicated in vivo","pmids":["32322062"],"is_preprint":false},{"year":2021,"finding":"PCK1 acetylates itself at its active site (self-acetylation) using acetyl-CoA as a substrate, independently of p300. Site-directed acetylation of K244 inside the active site renders the enzyme inactive, producing a ~3-fold decrease in kcat without changing Km, revealing acetyl-CoA as a regulatory ligand that controls PCK1 gluconeogenic activity.","method":"Protein crystallography, mass spectrometry, isothermal titration calorimetry, saturation-transfer difference NMR, molecular docking, in vitro acetylation assay, site-directed mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — reconstitution in vitro with structure, mutagenesis, and multiple biophysical methods in one study","pmids":["33334880"],"is_preprint":false},{"year":2024,"finding":"SR18292 increases PCK1 lysine acetylation (K91 mimicked by K91Q mutant), which reverses PCK1 catalytic direction to favor anaplerotic OAA synthesis from PEP, thereby supplying OAA to the TCA cycle, increasing lactate and glucose oxidation, and suppressing gluconeogenesis. Liver-specific expression of PCK1 K91Q ameliorates hyperglycemia in obese mice.","method":"In vitro enzyme activity assays with acetylation-mimetic mutant (K91Q), metabolic flux analysis, liver-specific viral expression in mice, pharmacological treatment with SR18292","journal":"Cell chemical biology","confidence":"High","confidence_rationale":"Tier 1 — mutagenesis plus in vivo rescue, mechanistic confirmation of reverse catalytic reaction","pmids":["39341205"],"is_preprint":false},{"year":2025,"finding":"Hypoxia induces JNK1/2-mediated phosphorylation of PCK1 at S151, which promotes physical interaction between PCK1 and cGAS. PCK1 associated with cGAS competitively consumes GTP (a shared substrate), thereby inhibiting GTP-dependent cGAS activation, suppressing STING-mediated immune cell recruitment, and promoting tumor immune evasion.","method":"Co-IP, in vitro GTP competition assays, phosphomimetic/phosphodeficient mutants, mouse tumor models with PCK1 blockade + anti-PD-1 combination, correlation in human breast cancer specimens","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 1/2 — biochemical reconstitution of competition, mutagenesis, and in vivo tumor models with multiple orthogonal approaches","pmids":["40048154"],"is_preprint":false},{"year":2021,"finding":"PCK1 deficiency causes oxaloacetate accumulation and increased de novo UTP synthesis, elevating UDP-GlcNAc and global O-GlcNAcylation. Simultaneously, PCK1 loss inactivates the AMPK–GFAT1 axis, further promoting UDP-GlcNAc synthesis. Elevated O-GlcNAcylation of CHK2 at Thr378 counteracts CHK2 stability and dimer formation, increasing CHK2-dependent Rb phosphorylation and HCC cell proliferation.","method":"PCK1 knockout cells, metabolic flux analysis, AMPK pathway knockdown/rescue, O-GlcNAc mass spectrometry, site-specific mutagenesis of CHK2 T378, liver-specific Pck1 knockout mouse HCC model","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (metabolomics, genetic rescue, site mutagenesis, in vivo model) in a single study","pmids":["33690219"],"is_preprint":false},{"year":2021,"finding":"PCK1 depletion increases O-GlcNAcylation of KAT5 (lysine acetyltransferase 5), which suppresses KAT5 ubiquitination and stabilizes it. Stabilized O-GlcNAc-KAT5 epigenetically activates TWIST1 via histone H4 acetylation and upregulates MMP9/MMP14 via c-Myc acetylation, promoting EMT and HCC metastasis.","method":"Gain/loss-of-function studies, Co-IP, ubiquitination assays, ChIP, O-GlcNAc site mapping by MS, in vivo lung metastasis in liver-specific Pck1-KO mice","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal assays (Co-IP, ChIP, ubiquitination, in vivo metastasis) demonstrating mechanistic chain","pmids":["34650217"],"is_preprint":false},{"year":2023,"finding":"PCK1 fuels the serine synthesis pathway to generate S-adenosylmethionine (SAM). SAM serves as a methyl donor for SUV39H1-catalyzed H3K9me3 modification at the S100A11 promoter, suppressing this oncogene. PCK1 deficiency reduces SAM, lowers H3K9me3 at S100A11, and induces oncogenic PI3K/AKT signaling via S100A11–AKT1 interaction.","method":"Metabolomic tracing of serine/SAM pathway, ChIP for H3K9me3, Co-IP of S100A11–AKT1, SAM supplementation rescue, S100A11 KO in vivo, PCK1 KO/OE hepatoma models","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 — multi-omics plus genetic rescue in vitro and in vivo, mechanistically linking PCK1 to epigenetic control via SAM","pmids":["37166978"],"is_preprint":false},{"year":2017,"finding":"CD8+ memory T cells upregulate PCK1 to drive gluconeogenesis-dependent glycogenesis. The resulting glycogen is channeled through glycogenolysis → glucose-6-phosphate → pentose phosphate pathway, generating NADPH that maintains reduced glutathione and limits ROS. Abrogation of the Pck1–glycogen–PPP axis impairs memory T-cell formation and maintenance.","method":"Pck1 KO and overexpression in T cells, metabolic flux analysis (glycogen, PPP intermediates, NADPH/GSH), mouse tumor immunotherapy models","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — genetic KO with defined metabolic readouts, rescue experiments, and in vivo immunotherapy validation","pmids":["29230018"],"is_preprint":false},{"year":2018,"finding":"Forced PCK1 expression in glucose-deprived liver cancer cells induces TCA cataplerosis, leading to energy crisis and oxidative stress that causes apoptosis. This pro-apoptotic effect requires PCK1 catalytic activity, as catalytic-dead mutants fail to recapitulate it. Replenishing α-ketoglutarate or inhibiting ROS blocks the PCK1-induced cell death.","method":"PCK1 overexpression with catalytic mutant controls, TCA metabolite profiling, ROS measurement, α-ketoglutarate rescue, xenograft tumor models","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1/2 — catalytic-dead mutagenesis plus metabolic rescue establishes enzymatic mechanism","pmids":["29335519"],"is_preprint":false},{"year":2019,"finding":"PCK1 upregulation in metastatic colorectal cancer cells drives pyrimidine nucleotide biosynthesis under hypoxia by supplying carbon via the gluconeogenic reaction, thereby supporting liver metastatic colonization and hypoxic growth.","method":"In vivo PDX selection for metastatic capacity, metabolomic profiling of pyrimidine intermediates, DHODH inhibition with leflunomide, PCK1 functional knockdown in metastasis models","journal":"eLife","confidence":"Medium","confidence_rationale":"Tier 2 — in vivo selection model with metabolomics and pharmacological inhibition, but detailed biochemical mechanism of carbon channeling not fully reconstituted","pmids":["31841108"],"is_preprint":false},{"year":2019,"finding":"PCK1 deficiency inactivates AMPK, suppresses p27Kip1 expression, and stimulates the CDK/Rb/E2F pathway, accelerating G1→S transition and hepatoma cell proliferation under glucose starvation. Conversely, PCK1 overexpression reduces ATP, enhances AMPK phosphorylation, and induces G1 arrest; AMPK knockdown reverses the PCK1-induced arrest.","method":"Gain/loss-of-function with AMPK inhibitor/activator, flow cytometry cell-cycle analysis, Western blotting of CDK/Rb/E2F/AMPK/p27, xenograft models","journal":"Journal of experimental & clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 — genetic rescue (AMPK KD reversal) and pharmacological confirmation with defined phenotypic readout","pmids":["30717766"],"is_preprint":false},{"year":2023,"finding":"SHP-1 phosphatase is recruited to the regulatory regions of the PCK1 gene, interacts with RNA polymerase II, and acts as a transcriptional coactivator for PCK1 expression. This recruitment is dependent on SHP-1's association with the transcription factor STAT5. Loss of SHP-1 or STAT5 reduces RNA Pol II at the PCK1 promoter, decreasing PCK1 mRNA and gluconeogenesis.","method":"Co-IP, ChIP-seq, luciferase reporter assay, gluconeogenesis assay, siRNA knockdown of SHP-1 and STAT5","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP-seq + Co-IP + functional gluconeogenesis rescue, single lab","pmids":["37595871"],"is_preprint":false},{"year":2015,"finding":"Insulin-induced Ser256 phosphorylation of FOXO1 triggers its translocation from nuclear speckles to the nuclear periphery, where FOXO1 forms a complex with EHMT2 histone methyltransferase. This complex induces repressive histone modifications at the PCK1 promoter and requires NUP98 for nuclear peripheral positioning, leading to transcriptional repression of PCK1.","method":"Live-cell imaging of endogenous phospho-FOXO1, Co-IP of FOXO1-EHMT2, ChIP of histone marks at PCK1 promoter, dominant-negative FOXO1 mutants, NUP98 interaction studies","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods establishing a new FOXO1 nuclear mechanism, single lab","pmids":["25736587"],"is_preprint":false},{"year":2016,"finding":"The oncoprotein HBXIP suppresses PCK1 expression by: (1) upregulating miR-135a, which targets the 3′UTR of FOXO1 mRNA, and (2) activating PI3K/Akt to phosphorylate FOXO1, causing its nuclear export. Both mechanisms reduce FOXO1-driven PCK1 transcription. Overexpression of PCK1 abolishes HBXIP-promoted hepatoma growth in vitro and in vivo.","method":"miR-135a reporter assay (3′UTR luciferase), FOXO1 phosphorylation/localization by Western blot and IF, PCK1 rescue overexpression, xenograft models","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 3 — mechanistic chain established with multiple assays but heavily reliant on overexpression/knockdown without site mutagenesis","pmids":["27609066"],"is_preprint":false},{"year":2023,"finding":"METTL3-mediated N6-methyladenosine (m6A) modification of PCK1 mRNA is induced during hepatic ischemia-reperfusion (I/R) injury, promoting PCK1 mRNA export and increased PCK1 expression. Hepatic-specific knockout of METTL3 reduces m6A on PCK1 transcript, decreases PCK1 expression, and worsens I/R injury, while PCK1 overexpression is protective.","method":"MeRIP-seq, hepatocyte-specific METTL3 KO mice, I/R mouse models, PCK1 inhibitor and overexpression, mRNA export assays","journal":"Hepatology","confidence":"Medium","confidence_rationale":"Tier 2 — MeRIP-seq mechanistically links METTL3 to PCK1 mRNA regulation in vivo, with genetic KO and functional rescue","pmids":["38085830"],"is_preprint":false},{"year":2023,"finding":"PCK1 loss in hepatoma cells increases global protein O-GlcNAcylation through two mechanisms: (1) oxaloacetate accumulation drives increased de novo UTP and UDP-GlcNAc synthesis, and (2) AMPK inactivation releases inhibitory GFAT1 phosphorylation. hnRNPA2B1 promotes HCC by binding PCK1 mRNA and reducing its m6A methylation, thereby decreasing PCK1 expression.","method":"RNA immunoprecipitation (RIP), MeRIP assay for m6A, CRISPR-Cas9 KO of hnRNPA2B1, RNA-seq, subcutaneous and orthotopic HCC mouse models","journal":"Journal of translational medicine","confidence":"Medium","confidence_rationale":"Tier 2 — RIP and MeRIP establish direct binding and m6A mechanism; in vivo rescue confirms functional dependence on PCK1","pmids":["38017546"],"is_preprint":false},{"year":2005,"finding":"Loss of PCK1 in myeloid cells (macrophage-specific Pck1 deletion) reduces 13C labeling of TCA intermediates (citrate, malate) from [U-13C]glucose, increases 13C-lactate labeling, and elevates ROS, shifting macrophages toward a pro-inflammatory M1 phenotype with increased TNFα, IL-1β, and IL-6 expression.","method":"Myeloid-specific Pck1 KO mice, stable isotopomer MS with [U-13C]glucose in BMDM, ROS measurement, cytokine ELISA","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — isotope tracing in genetic KO macrophages directly links PCK1 to macrophage metabolic reprogramming and inflammatory phenotype","pmids":["29317502"],"is_preprint":false},{"year":2016,"finding":"A single amino acid substitution Met139Leu (c.A2456C SNP) in pig PCK1 reduces kcat in the glyceroneogenic direction, enhances kcat in the anaplerotic direction, and reduces ability of the enzyme to be acetylated, increasing its susceptibility to ubiquitin-proteasome degradation. The substitution results in ~30% lower glucose and ~9% lower lipid production in cell cultures.","method":"Recombinant enzyme kinetic assays (kcat, Km in both directions), acetylation assay, cell culture metabolic flux measurement, pig phenotype association","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1 — in vitro enzymatic characterization with defined mutant, plus post-translational modification consequence, single lab","pmids":["26792594"],"is_preprint":false},{"year":2023,"finding":"Kidney-specific PCK1 deletion in mice causes hyperchloremic metabolic acidosis with reduced ammoniagenesis, glycosuria, lactaturia, and decreased ATP generation. PCK1 is required for proximal tubule energy production and acid-base control; its loss increases tubular injury during acidosis, while overexpression protects against proteinuric renal disease.","method":"Kidney-specific PCK1 KO and knockin (PAX8 promoter), metabolic phenotyping (creatinine clearance, albuminuria, glycosuria), ATP measurement, acid-base challenge experiments","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 — tissue-specific genetic models with defined functional readouts (acid-base, energy, injury), single lab","pmids":["37102687"],"is_preprint":false},{"year":2020,"finding":"PCK1 loss in HCC deficiency activates the RhoA/PI3K/AKT pathway by increasing intracellular GTP levels, stimulates paracrine secretion of PDGF-AA, and promotes hepatic stellate cell activation and liver fibrosis. Treatment with RhoA and AKT inhibitors or silencing of RhoA/AKT1 alleviates MAFLD progression in PCK1-deficient mice.","method":"Liver-specific Pck1 KO mice, intracellular GTP measurement, PDGF-AA ELISA, hepatic stellate cell co-culture, RhoA/AKT inhibitor treatment, AAV-mediated PCK1 rescue","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 — GTP measurement mechanistically links PCK1 substrate consumption to RhoA/PI3K pathway; genetic and pharmacological rescue in vivo","pmids":["36918564"],"is_preprint":false},{"year":2024,"finding":"PCK1 enhances PYGL phosphorylation in cervical cancer stem cells, promoting glycogen breakdown and channeling glucose-6-phosphate into the pentose phosphate pathway, increasing NADPH production and ROS clearance, thereby promoting chemoresistance.","method":"siRNA knockdown of PCK1, PYGL, GYS1 in HCC94/CaSki cells, glycogen measurement, LC-MS of PPP intermediates, NADPH/NADP+ ratio, NSG mouse tumor growth assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — direct biochemical measurement of PPP/NADPH with genetic perturbations and in vivo tumor model","pmids":["38871968"],"is_preprint":false},{"year":2024,"finding":"PRRSV infection activates AKT, which in turn activates PCK1. Activated PCK1 phosphorylates INSIG proteins (INSIGs), causing their degradation and releasing SCAP–SREBP complexes to the nucleus to activate lipid biosynthesis; ROS produced by PRRSV are required for AKT activation in this axis.","method":"Pharmacological inhibitors of AKT and PCK1, INSIG protein level monitoring by Western blot, SREBP nuclear translocation assay, ROS scavenger treatment, MARC-145 cell infection model","journal":"International journal of biological macromolecules","confidence":"Low","confidence_rationale":"Tier 3 — mechanistic evidence relies primarily on pharmacological inhibitors and Western blot in cell culture, limited genetic validation","pmids":["39433189"],"is_preprint":false},{"year":2019,"finding":"Rev-erbα directly binds a RevRE site at −325 to −320 bp in the PCK1 promoter to trans-repress PCK1 transcription, reducing hepatic Pck1 mRNA and protein levels and lowering fasting plasma glucose in diabetic mice.","method":"Luciferase reporter assay with promoter deletions, gel-shift (EMSA) with RevRE site probe, ChIP assay in hepatoma cells, SR9009 (Rev-erbα agonist) treatment in WT and STZ diabetic mice","journal":"Pharmacological research","confidence":"Medium","confidence_rationale":"Tier 2 — EMSA + ChIP + luciferase + in vivo pharmacology establish direct promoter binding and functional consequence","pmids":["30639375"],"is_preprint":false},{"year":2023,"finding":"PCK1 antagonizes CRC cell growth by inactivating UBAP2L phosphorylation at serine 454, thereby enhancing autophagy flux and suppressing tumor proliferation in vitro and in vivo.","method":"Overexpression and knockdown of PCK1, phosphoproteomic identification of UBAP2L pS454, autophagy flux assay (LC3, p62), xenograft models","journal":"Cancer cell international","confidence":"Low","confidence_rationale":"Tier 3 — phosphosite identified but kinase-substrate relationship not biochemically reconstituted; single lab","pmids":["37062825"],"is_preprint":false},{"year":2004,"finding":"The −232C>G promoter SNP of human PCK1 lies within a cis-acting element required for basal and cAMP-mediated transcription. The −232G allele in luciferase reporter assays shows increased basal expression and loss of insulin-mediated downregulation, establishing that this element is required for insulin suppression of PCK1 transcription.","method":"Luciferase reporter gene assay in three cell lines with −232C vs −232G constructs, insulin suppression experiments","journal":"The Journal of clinical endocrinology and metabolism","confidence":"Medium","confidence_rationale":"Tier 2 — functional reporter assay in multiple cell lines with specific promoter element directly tested","pmids":["14764811"],"is_preprint":false},{"year":2023,"finding":"PCK1 overexpression in proximal tubules suppresses HK2 upregulation (rate-limiting enzyme of glycolysis), thereby blocking excess glycolysis. PCK1 overexpression preserves mitoribosomal function and prevents tubular fibrosis in diabetic nephropathy; conversely, PCK1 CKO mice develop mitoribosomal defects and renal fibrosis.","method":"PT-specific Pck1 TG and CKO mice, STZ diabetic model, Western blot for mitoribosomes and HK2, type IV collagen deposition, albuminuria measurement","journal":"Journal of the American Society of Nephrology","confidence":"Medium","confidence_rationale":"Tier 2 — bidirectional genetic models (TG and CKO) in disease model with defined molecular and histological readouts","pmids":["37199399"],"is_preprint":false}],"current_model":"PCK1 (cytosolic PEPCK) is a multifunctional enzyme that catalyzes the GTP-dependent conversion of oxaloacetate to phosphoenolpyruvate for gluconeogenesis, but is also regulated post-translationally: AKT-mediated Ser90 phosphorylation redirects PCK1 to the ER where it acts as a GTP-dependent protein kinase phosphorylating INSIG1/2 to activate SREBP-driven lipogenesis; acetyl-CoA binds the active site and drives self-acetylation (notably at K244/K91) to inhibit gluconeogenic activity and favor anaplerotic OAA synthesis; JNK1/2-mediated S151 phosphorylation promotes PCK1–cGAS interaction that competitively consumes GTP to suppress innate immune signaling; and through its enzymatic activity PCK1 controls TCA cataplerosis, glycogenesis/PPP flux for NADPH and ROS homeostasis, SAM production for H3K9 methylation, and O-GlcNAcylation levels, collectively regulating metabolism, epigenetics, cell-cycle progression via AMPK/p27, and immune evasion in cancer and normal physiology."},"narrative":{"teleology":[{"year":2004,"claim":"Identification of a cis-regulatory element at −232 in the PCK1 promoter established that a single nucleotide variant governs basal transcription and insulin-mediated repression, providing the first functional dissection of how insulin silences PCK1 at the promoter level.","evidence":"Luciferase reporter assays with −232C vs −232G constructs in multiple cell lines with insulin treatment","pmids":["14764811"],"confidence":"Medium","gaps":["Chromatin context of the −232 element not examined","Trans-acting factor binding this element not identified","No in vivo confirmation of allele-specific expression"]},{"year":2015,"claim":"Discovery that insulin-phosphorylated FOXO1 relocates to the nuclear periphery and recruits the histone methyltransferase EHMT2, establishing repressive chromatin at the PCK1 promoter, revealed a spatial–epigenetic mechanism of PCK1 transcriptional silencing beyond simple FOXO1 nuclear exclusion.","evidence":"Live-cell imaging of endogenous phospho-FOXO1, Co-IP of FOXO1–EHMT2, ChIP of histone marks at PCK1 promoter, NUP98 interaction studies","pmids":["25736587"],"confidence":"Medium","gaps":["Not independently confirmed by a second lab","Contribution relative to canonical FOXO1 nuclear export not quantified","Whether this mechanism operates genome-wide or is PCK1-specific is unclear"]},{"year":2017,"claim":"Demonstration that CD8+ memory T cells upregulate PCK1 to fuel a gluconeogenesis→glycogenesis→PPP axis for NADPH and ROS control established a non-hepatic, immunological role for PCK1 and linked its catalytic activity to adaptive immune memory.","evidence":"T-cell-specific Pck1 KO and overexpression, 13C metabolic flux analysis, NADPH/GSH measurement, mouse tumor immunotherapy models","pmids":["29230018"],"confidence":"High","gaps":["Whether PCK1 is required in all memory T-cell subsets is unknown","Upstream signals inducing PCK1 in memory T cells not fully defined","Relative contribution of glycogen vs direct PPP flux not dissected"]},{"year":2018,"claim":"Showing that forced PCK1 expression in glucose-deprived hepatoma cells causes lethal TCA cataplerosis, energy crisis, and ROS-mediated apoptosis—dependent on catalytic activity—established PCK1 as a context-dependent tumor suppressor whose metabolic drain is cytotoxic.","evidence":"PCK1 overexpression with catalytic-dead mutant controls, TCA metabolite profiling, α-ketoglutarate rescue, xenograft models","pmids":["29335519"],"confidence":"High","gaps":["Threshold of cataplerosis required for apoptosis not quantified","Relevance to non-hepatic tumors not tested in this study","Downstream ROS sensors mediating apoptosis not identified"]},{"year":2019,"claim":"Linking PCK1 loss to AMPK inactivation, p27 suppression, and accelerated G1→S transition provided a direct mechanism by which metabolic sensing through PCK1 controls cell-cycle progression in hepatoma cells.","evidence":"PCK1 gain/loss-of-function with AMPK inhibitor/activator, flow cytometry, CDK/Rb/E2F pathway analysis, xenograft models","pmids":["30717766"],"confidence":"Medium","gaps":["Whether the ATP drop is the sole AMPK-activating signal is unresolved","Contribution of other AMPK substrates beyond p27 not tested","Single cancer type studied"]},{"year":2019,"claim":"Identification of PCK1 upregulation in metastatic colorectal cancer cells driving pyrimidine nucleotide biosynthesis under hypoxia revealed that the gluconeogenic reaction can supply carbon for nucleotide synthesis in non-hepatic tumors, expanding PCK1's oncogenic role.","evidence":"PDX in vivo selection, metabolomic profiling of pyrimidine intermediates, DHODH inhibition, PCK1 knockdown in metastasis models","pmids":["31841108"],"confidence":"Medium","gaps":["Exact carbon routing from PEP to pyrimidines not traced isotopically at single-step resolution","Whether this is specific to liver metastases or generalizable is unclear","No structural or enzymatic reconstitution"]},{"year":2020,"claim":"The landmark finding that AKT phosphorylates PCK1 at Ser90, causing ER translocation and a moonlighting protein kinase activity toward INSIG1/2, fundamentally redefined PCK1 as a signal-regulated enzyme with dual catalytic identities—metabolic and signaling.","evidence":"In vitro kinase assays, site-directed mutagenesis, Co-IP, subcellular fractionation, ER translocation imaging, xenograft and human HCC correlation","pmids":["32322062"],"confidence":"High","gaps":["Full substrate scope of PCK1 kinase activity beyond INSIGs not surveyed","Structural basis for GTP-dependent kinase mechanism not resolved","Whether kinase activity occurs in non-tumor tissues is unknown"]},{"year":2021,"claim":"Crystal-structure-guided discovery that acetyl-CoA binds the PCK1 active site and drives self-acetylation at K244, inactivating the enzyme, established a direct feedback mechanism by which acetyl-CoA availability controls gluconeogenic flux independently of acetyltransferases.","evidence":"X-ray crystallography, mass spectrometry, ITC, STD-NMR, in vitro acetylation assay, site-directed mutagenesis","pmids":["33334880"],"confidence":"High","gaps":["Physiological acetyl-CoA concentrations at which self-acetylation becomes rate-limiting not defined","Whether deacetylases counteract this in real time in vivo is unclear","Impact on protein kinase activity not tested"]},{"year":2021,"claim":"Comprehensive metabolic tracing showed PCK1 loss causes OAA accumulation, increased UDP-GlcNAc, and global O-GlcNAcylation—including CHK2 at T378, which destabilizes CHK2 and drives Rb phosphorylation and proliferation—establishing a PCK1→O-GlcNAc→cell-cycle axis in HCC.","evidence":"PCK1 KO cells, metabolic flux analysis, AMPK–GFAT1 epistasis, O-GlcNAc MS, CHK2 T378 mutagenesis, liver-specific Pck1 KO HCC mice","pmids":["33690219","34650217"],"confidence":"High","gaps":["Complete O-GlcNAc proteome altered by PCK1 loss not catalogued","Whether O-GlcNAcylation changes are reversible upon PCK1 restoration in established tumors is unknown","Relative contribution of OAA-driven vs AMPK-driven UDP-GlcNAc increase not quantified"]},{"year":2023,"claim":"Discovery that PCK1 fuels serine/SAM biosynthesis and thereby sustains SUV39H1-catalyzed H3K9me3 at the S100A11 promoter linked PCK1 catalytic activity to epigenetic gene silencing and identified a metabolic–epigenetic tumor-suppressive circuit.","evidence":"Metabolomic tracing of serine/SAM, ChIP for H3K9me3, Co-IP of S100A11–AKT1, SAM rescue, S100A11 KO in vivo, PCK1 KO/OE hepatoma models","pmids":["37166978"],"confidence":"High","gaps":["Genome-wide H3K9me3 changes upon PCK1 loss not reported","Whether other SAM-consuming methyltransferases are similarly affected is unclear","Direct isotope tracing from PEP to SAM not shown at single-step resolution"]},{"year":2023,"claim":"Kidney-specific PCK1 deletion caused hyperchloremic metabolic acidosis, reduced ammoniagenesis, and impaired tubular ATP production, establishing PCK1 as essential for renal acid-base homeostasis and energy metabolism beyond its hepatic role.","evidence":"Kidney-specific PCK1 KO and knockin mice, metabolic phenotyping, ATP measurement, acid-base challenge","pmids":["37102687","37199399"],"confidence":"Medium","gaps":["Whether PCK1 compensates for PCK2 loss in kidney is not tested","Mechanism linking PCK1 to ammoniagenesis not fully delineated","Long-term renal outcomes of PCK1 overexpression not characterized"]},{"year":2024,"claim":"Showing that K91 acetylation reverses PCK1's catalytic direction to favor anaplerotic OAA synthesis from PEP, and that liver-specific K91Q expression ameliorates hyperglycemia in obese mice, revealed a post-translational switch that repurposes PCK1 from catabolic to anabolic function in vivo.","evidence":"In vitro enzyme activity with K91Q mimetic, metabolic flux analysis, liver-specific viral expression in obese mice, SR18292 pharmacology","pmids":["39341205"],"confidence":"High","gaps":["Whether K91 acetylation and K244 acetylation cooperate or are mutually exclusive is unresolved","Deacetylase(s) responsible for reversing K91 acetylation in vivo not identified","Effect of reverse reaction on non-hepatic tissues not explored"]},{"year":2025,"claim":"Identification of JNK1/2-mediated S151 phosphorylation driving PCK1–cGAS association and competitive GTP consumption revealed a non-enzymatic, immune-suppressive function of PCK1 that enables tumor immune evasion via cGAS–STING inhibition.","evidence":"Co-IP, in vitro GTP competition assays, phosphomimetic/phosphodeficient mutants, mouse tumor models with anti-PD-1, human breast cancer specimen correlation","pmids":["40048154"],"confidence":"High","gaps":["Whether PCK1–cGAS interaction occurs in non-tumor immune contexts is unknown","Structural basis of the PCK1–cGAS interface not determined","Relative contribution of GTP depletion vs direct cGAS inhibition not fully separated"]},{"year":null,"claim":"Major open questions include the full substrate scope of PCK1's protein kinase activity, the structural basis for its dual catalytic (metabolic and kinase) functions, how the multiple post-translational modifications (Ser90, S151, K91, K244 phosphorylation/acetylation) are hierarchically coordinated in vivo, and whether PCK1's immune-modulatory roles extend beyond the tumor microenvironment.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal structure of phospho-Ser90 ER-localized PCK1 in kinase conformation","Integrated PTM code governing catalytic direction not mapped","Non-cancer immune roles of PCK1 largely unexplored"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,3,21]},{"term_id":"GO:0016853","term_label":"isomerase activity","supporting_discovery_ids":[1,2]},{"term_id":"GO:0016829","term_label":"lyase activity","supporting_discovery_ids":[2,8]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,8]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0,21]}],"pathway":[{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[1,2,7,8,9,17,18]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4,10,19,21]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3,7,16]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[6]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4,10]}],"complexes":[],"partners":["INSIG1","INSIG2","AKT1","CGAS","CHK2","PYGL","UBAP2L","SCAP"],"other_free_text":[]},"mechanistic_narrative":"PCK1 (cytosolic phosphoenolpyruvate carboxykinase) is a GTP-dependent enzyme whose canonical activity—conversion of oxaloacetate to phosphoenolpyruvate—positions it as a master regulator of carbon flux through gluconeogenesis, TCA cataplerosis, glycogenesis, the pentose phosphate pathway, and one-carbon/SAM metabolism, with consequences for NADPH/ROS homeostasis, epigenetic H3K9 trimethylation, O-GlcNAcylation, and cell-cycle control via AMPK/p27 [PMID:29230018, PMID:33690219, PMID:37166978, PMID:30717766]. Beyond its metabolic activity, AKT-mediated Ser90 phosphorylation redirects PCK1 to the endoplasmic reticulum where it functions as a protein kinase, phosphorylating INSIG1/2 to release SCAP–SREBP and activate lipogenesis [PMID:32322062]. Acetyl-CoA binding at the active site drives PCK1 self-acetylation (K244, K91), which inactivates the gluconeogenic reaction and can reverse catalytic directionality to favor anaplerotic oxaloacetate synthesis [PMID:33334880, PMID:39341205]. JNK1/2-mediated S151 phosphorylation promotes PCK1–cGAS interaction and competitive GTP consumption, suppressing cGAS–STING innate immune signaling and enabling tumor immune evasion [PMID:40048154]."},"prefetch_data":{"uniprot":{"accession":"P35558","full_name":"Phosphoenolpyruvate carboxykinase, cytosolic [GTP]","aliases":["Serine-protein kinase PCK1"],"length_aa":622,"mass_kda":69.2,"function":"Cytosolic phosphoenolpyruvate carboxykinase that catalyzes the reversible decarboxylation and phosphorylation of oxaloacetate (OAA) and acts as the rate-limiting enzyme in gluconeogenesis (PubMed:24863970, PubMed:26971250, PubMed:28216384, PubMed:30193097). Regulates cataplerosis and anaplerosis, the processes that control the levels of metabolic intermediates in the citric acid cycle (PubMed:24863970, PubMed:26971250, PubMed:28216384, PubMed:30193097). At low glucose levels, it catalyzes the cataplerotic conversion of oxaloacetate to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle (PubMed:30193097). At high glucose levels, it catalyzes the anaplerotic conversion of phosphoenolpyruvate to oxaloacetate (PubMed:30193097). Acts as a regulator of formation and maintenance of memory CD8(+) T-cells: up-regulated in these cells, where it generates phosphoenolpyruvate, via gluconeogenesis (By similarity). The resultant phosphoenolpyruvate flows to glycogen and pentose phosphate pathway, which is essential for memory CD8(+) T-cells homeostasis (By similarity). In addition to the phosphoenolpyruvate carboxykinase activity, also acts as a protein kinase when phosphorylated at Ser-90: phosphorylation at Ser-90 by AKT1 reduces the binding affinity to oxaloacetate and promotes an atypical serine protein kinase activity using GTP as donor (PubMed:32322062). The protein kinase activity regulates lipogenesis: upon phosphorylation at Ser-90, translocates to the endoplasmic reticulum and catalyzes phosphorylation of INSIG proteins (INSIG1 and INSIG2), thereby disrupting the interaction between INSIG proteins and SCAP and promoting nuclear translocation of SREBP proteins (SREBF1/SREBP1 or SREBF2/SREBP2) and subsequent transcription of downstream lipogenesis-related genes (PubMed:32322062)","subcellular_location":"Cytoplasm, cytosol; Endoplasmic reticulum","url":"https://www.uniprot.org/uniprotkb/P35558/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/PCK1","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/PCK1","total_profiled":1310},"omim":[{"mim_id":"618964","title":"REQUIRED FOR MEIOTIC NUCLEAR DIVISION 5 HOMOLOG A; RMND5A","url":"https://www.omim.org/entry/618964"},{"mim_id":"617758","title":"ZINC FINGER PROTEIN 692; ZNF692","url":"https://www.omim.org/entry/617758"},{"mim_id":"617699","title":"GID COMPLEX, SUBUNIT 4; GID4","url":"https://www.omim.org/entry/617699"},{"mim_id":"614168","title":"PHOSPHOENOLPYRUVATE CARBOXYKINASE 1, SOLUBLE; PCK1","url":"https://www.omim.org/entry/614168"},{"mim_id":"614095","title":"PHOSPHOENOLPYRUVATE CARBOXYKINASE 2, MITOCHONDRIAL; PCK2","url":"https://www.omim.org/entry/614095"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"kidney","ntpm":403.6},{"tissue":"liver","ntpm":643.7}],"url":"https://www.proteinatlas.org/search/PCK1"},"hgnc":{"alias_symbol":["PEPCK-C"],"prev_symbol":[]},"alphafold":{"accession":"P35558","domains":[{"cath_id":"3.40.449.10","chopping":"12-65_92-247","consensus_level":"high","plddt":97.5506,"start":12,"end":247},{"cath_id":"3.90.228.20","chopping":"262-331_418-622","consensus_level":"high","plddt":97.0031,"start":262,"end":622},{"cath_id":"2.170.8.10","chopping":"334-413","consensus_level":"medium","plddt":96.8014,"start":334,"end":413}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P35558","model_url":"https://alphafold.ebi.ac.uk/files/AF-P35558-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P35558-F1-predicted_aligned_error_v6.png","plddt_mean":96.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=PCK1","jax_strain_url":"https://www.jax.org/strain/search?query=PCK1"},"sequence":{"accession":"P35558","fasta_url":"https://rest.uniprot.org/uniprotkb/P35558.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P35558/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P35558"}},"corpus_meta":[{"pmid":"32322062","id":"PMC_32322062","title":"The gluconeogenic enzyme PCK1 phosphorylates INSIG1/2 for lipogenesis.","date":"2020","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/32322062","citation_count":290,"is_preprint":false},{"pmid":"29230018","id":"PMC_29230018","title":"A Pck1-directed glycogen metabolic program regulates formation and maintenance of memory CD8+ T cells.","date":"2017","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29230018","citation_count":174,"is_preprint":false},{"pmid":"29335519","id":"PMC_29335519","title":"Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma.","date":"2018","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/29335519","citation_count":163,"is_preprint":false},{"pmid":"17709878","id":"PMC_17709878","title":"PCK1 and PCK2 as candidate diabetes and obesity genes.","date":"2007","source":"Cell biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/17709878","citation_count":162,"is_preprint":false},{"pmid":"36918564","id":"PMC_36918564","title":"Deficiency of gluconeogenic enzyme PCK1 promotes metabolic-associated fatty liver disease through PI3K/AKT/PDGF axis activation in male mice.","date":"2023","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/36918564","citation_count":126,"is_preprint":false},{"pmid":"18443203","id":"PMC_18443203","title":"Pck1 gene silencing in the liver improves glycemia control, insulin sensitivity, and dyslipidemia in db/db mice.","date":"2008","source":"Diabetes","url":"https://pubmed.ncbi.nlm.nih.gov/18443203","citation_count":111,"is_preprint":false},{"pmid":"33690219","id":"PMC_33690219","title":"Gluconeogenic enzyme PCK1 deficiency promotes CHK2 O-GlcNAcylation and hepatocellular carcinoma growth upon glucose deprivation.","date":"2021","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/33690219","citation_count":101,"is_preprint":false},{"pmid":"15046742","id":"PMC_15046742","title":"Disregulated glyceroneogenesis: PCK1 as a candidate diabetes and obesity gene.","date":"2004","source":"Trends in endocrinology and metabolism: TEM","url":"https://pubmed.ncbi.nlm.nih.gov/15046742","citation_count":99,"is_preprint":false},{"pmid":"29608893","id":"PMC_29608893","title":"Circular RNA circC3P1 suppresses hepatocellular carcinoma growth and metastasis through miR-4641/PCK1 pathway.","date":"2018","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/29608893","citation_count":91,"is_preprint":false},{"pmid":"23466304","id":"PMC_23466304","title":"PEPCK-M expression in mouse liver potentiates, not replaces, PEPCK-C mediated gluconeogenesis.","date":"2013","source":"Journal of hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/23466304","citation_count":87,"is_preprint":false},{"pmid":"20124556","id":"PMC_20124556","title":"Phosphoenolpyruvate carboxykinase (Pck1) helps regulate the triglyceride/fatty acid cycle and development of insulin resistance in mice.","date":"2010","source":"Journal of lipid research","url":"https://pubmed.ncbi.nlm.nih.gov/20124556","citation_count":81,"is_preprint":false},{"pmid":"31841108","id":"PMC_31841108","title":"PCK1 and DHODH drive colorectal cancer liver metastatic colonization and hypoxic growth by promoting nucleotide synthesis.","date":"2019","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/31841108","citation_count":80,"is_preprint":false},{"pmid":"9224895","id":"PMC_9224895","title":"Sequence and promoter regulation of the PCK1 gene encoding phosphoenolpyruvate carboxykinase of the fungal pathogen Candida albicans.","date":"1997","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/9224895","citation_count":80,"is_preprint":false},{"pmid":"38718791","id":"PMC_38718791","title":"A 5:2 intermittent fasting regimen ameliorates NASH and fibrosis and blunts HCC development via hepatic PPARα and PCK1.","date":"2024","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/38718791","citation_count":72,"is_preprint":false},{"pmid":"30717766","id":"PMC_30717766","title":"PCK1 negatively regulates cell cycle progression and hepatoma cell proliferation via the AMPK/p27Kip1 axis.","date":"2019","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/30717766","citation_count":69,"is_preprint":false},{"pmid":"29768256","id":"PMC_29768256","title":"Overexpression of PCK1 Gene Antagonizes Hepatocellular Carcinoma Through the Activation of Gluconeogenesis and Suppression of Glycolysis Pathways.","date":"2018","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29768256","citation_count":65,"is_preprint":false},{"pmid":"24863970","id":"PMC_24863970","title":"Three rare diseases in one Sib pair: RAI1, PCK1, GRIN2B mutations associated with Smith-Magenis Syndrome, cytosolic PEPCK deficiency and NMDA receptor glutamate insensitivity.","date":"2014","source":"Molecular genetics and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/24863970","citation_count":56,"is_preprint":false},{"pmid":"1327878","id":"PMC_1327878","title":"Regulatory regions in the yeast FBP1 and PCK1 genes.","date":"1992","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/1327878","citation_count":52,"is_preprint":false},{"pmid":"17440948","id":"PMC_17440948","title":"Candidate gene association study of insulin signaling genes and Alzheimer's disease: evidence for SOS2, PCK1, and PPARgamma as susceptibility loci.","date":"2007","source":"American journal of medical genetics. Part B, Neuropsychiatric genetics : the official publication of the International Society of Psychiatric Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/17440948","citation_count":52,"is_preprint":false},{"pmid":"34650217","id":"PMC_34650217","title":"O-GlcNAc modified-TIP60/KAT5 is required for PCK1 deficiency-induced HCC metastasis.","date":"2021","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/34650217","citation_count":49,"is_preprint":false},{"pmid":"37166978","id":"PMC_37166978","title":"Gluconeogenic enzyme PCK1 supports S-adenosylmethionine biosynthesis and promotes H3K9me3 modification to suppress hepatocellular carcinoma progression.","date":"2023","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/37166978","citation_count":48,"is_preprint":false},{"pmid":"27609066","id":"PMC_27609066","title":"The oncoprotein HBXIP suppresses gluconeogenesis through modulating PCK1 to enhance the growth of hepatoma cells.","date":"2016","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/27609066","citation_count":47,"is_preprint":false},{"pmid":"14764811","id":"PMC_14764811","title":"Promoter polymorphism in PCK1 (phosphoenolpyruvate carboxykinase gene) associated with type 2 diabetes mellitus.","date":"2004","source":"The Journal of clinical endocrinology and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/14764811","citation_count":45,"is_preprint":false},{"pmid":"30619751","id":"PMC_30619751","title":"PCK1 Downregulation Promotes TXNRD1 Expression and Hepatoma Cell Growth via the Nrf2/Keap1 Pathway.","date":"2018","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/30619751","citation_count":45,"is_preprint":false},{"pmid":"37102687","id":"PMC_37102687","title":"PCK1 is a key regulator of metabolic and mitochondrial functions in renal tubular cells.","date":"2023","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/37102687","citation_count":42,"is_preprint":false},{"pmid":"7854322","id":"PMC_7854322","title":"Identification and characterization of regulatory elements in the phosphoenolpyruvate carboxykinase gene PCK1 of Saccharomyces cerevisiae.","date":"1995","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/7854322","citation_count":41,"is_preprint":false},{"pmid":"29317502","id":"PMC_29317502","title":"Macrophages with a deletion of the phosphoenolpyruvate carboxykinase 1 (Pck1) gene have a more proinflammatory phenotype.","date":"2018","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29317502","citation_count":38,"is_preprint":false},{"pmid":"30913382","id":"PMC_30913382","title":"Induction of Inflammatory Responses in Human Bronchial Epithelial Cells by Pb2+-Containing Model PM2.5 Particles via Downregulation of a Novel Long Noncoding RNA lnc-PCK1-2:1.","date":"2019","source":"Environmental science & technology","url":"https://pubmed.ncbi.nlm.nih.gov/30913382","citation_count":38,"is_preprint":false},{"pmid":"37199399","id":"PMC_37199399","title":"PCK1 Protects against Mitoribosomal Defects in Diabetic Nephropathy in Mouse Models.","date":"2023","source":"Journal of the American Society of Nephrology : JASN","url":"https://pubmed.ncbi.nlm.nih.gov/37199399","citation_count":37,"is_preprint":false},{"pmid":"21731709","id":"PMC_21731709","title":"Insulin-regulated Srebp-1c and Pck1 mRNA expression in primary hepatocytes from zucker fatty but not lean rats is affected by feeding conditions.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21731709","citation_count":35,"is_preprint":false},{"pmid":"32809272","id":"PMC_32809272","title":"A newly discovered role of metabolic enzyme PCK1 as a protein kinase to promote cancer lipogenesis.","date":"2020","source":"Cancer communications (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/32809272","citation_count":34,"is_preprint":false},{"pmid":"24498240","id":"PMC_24498240","title":"Rho1 GTPase and PKC ortholog Pck1 are upstream activators of the cell integrity MAPK pathway in fission yeast.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24498240","citation_count":33,"is_preprint":false},{"pmid":"32280238","id":"PMC_32280238","title":"PCK1 Regulates Glycolysis and Tumor Progression in Clear Cell Renal Cell Carcinoma Through LDHA.","date":"2020","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/32280238","citation_count":32,"is_preprint":false},{"pmid":"28216384","id":"PMC_28216384","title":"Novel homozygous PCK1 mutation causing cytosolic phosphoenolpyruvate carboxykinase deficiency presenting as childhood hypoglycemia, an abnormal pattern of urine metabolites and liver dysfunction.","date":"2017","source":"Molecular genetics and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/28216384","citation_count":28,"is_preprint":false},{"pmid":"23544081","id":"PMC_23544081","title":"Androgen inhibits abdominal fat accumulation and negatively regulates the PCK1 gene in male chickens.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23544081","citation_count":28,"is_preprint":false},{"pmid":"33045988","id":"PMC_33045988","title":"Comparative transcriptome analysis reveals that PCK1 is a potential gene affecting IMF deposition in buffalo.","date":"2020","source":"BMC genomics","url":"https://pubmed.ncbi.nlm.nih.gov/33045988","citation_count":27,"is_preprint":false},{"pmid":"29614212","id":"PMC_29614212","title":"The key gluconeogenic gene PCK1 is crucial for virulence of Botrytis cinerea via initiating its conidial germination and host penetration.","date":"2018","source":"Environmental microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/29614212","citation_count":27,"is_preprint":false},{"pmid":"11016849","id":"PMC_11016849","title":"Differences in regulation of yeast gluconeogenesis revealed by Cat8p-independent activation of PCK1 and FBP1 genes in Kluyveromyces lactis.","date":"2000","source":"Molecular & general genetics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/11016849","citation_count":27,"is_preprint":false},{"pmid":"28816409","id":"PMC_28816409","title":"Genistein represses PEPCK-C expression in an insulin-independent manner in HepG2 cells and in alloxan-induced diabetic mice.","date":"2017","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28816409","citation_count":27,"is_preprint":false},{"pmid":"21647250","id":"PMC_21647250","title":"Berberine regulated Gck, G6pc, Pck1 and Srebp-1c expression and activated AMP-activated protein kinase in primary rat hepatocytes.","date":"2011","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/21647250","citation_count":27,"is_preprint":false},{"pmid":"30639375","id":"PMC_30639375","title":"Rev-erbα activation down-regulates hepatic Pck1 enzyme to lower plasma glucose in mice.","date":"2019","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/30639375","citation_count":26,"is_preprint":false},{"pmid":"19725958","id":"PMC_19725958","title":"The promoter polymorphism -232C/G of the PCK1 gene is associated with type 2 diabetes in a UK-resident South Asian population.","date":"2009","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19725958","citation_count":25,"is_preprint":false},{"pmid":"8490617","id":"PMC_8490617","title":"cDNA sequence and localization of polymorphic human cytosolic phosphoenolpyruvate carboxykinase gene (PCK1) to chromosome 20, band q13.31: PCK1 is not tightly linked to maturity-onset diabetes of the young.","date":"1993","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/8490617","citation_count":25,"is_preprint":false},{"pmid":"21519922","id":"PMC_21519922","title":"Retinoids induced Pck1 expression and attenuated insulin-mediated suppression of its expression via activation of retinoic acid receptor in primary rat hepatocytes.","date":"2011","source":"Molecular and cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21519922","citation_count":24,"is_preprint":false},{"pmid":"8325643","id":"PMC_8325643","title":"Phosphoenolpyruvate carboxykinase (GTP): characterization of the human PCK1 gene and localization distal to MODY on chromosome 20.","date":"1993","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8325643","citation_count":24,"is_preprint":false},{"pmid":"17678617","id":"PMC_17678617","title":"Liver lipid molecules induce PEPCK-C gene transcription and attenuate insulin action.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17678617","citation_count":24,"is_preprint":false},{"pmid":"37013052","id":"PMC_37013052","title":"PCK1 dysregulation in cancer: Metabolic reprogramming, oncogenic activation, and therapeutic opportunities.","date":"2022","source":"Genes & diseases","url":"https://pubmed.ncbi.nlm.nih.gov/37013052","citation_count":23,"is_preprint":false},{"pmid":"25736587","id":"PMC_25736587","title":"Translocation of forkhead box O1 to the nuclear periphery induces histone modifications that regulate transcriptional repression of PCK1 in HepG2 cells.","date":"2015","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/25736587","citation_count":21,"is_preprint":false},{"pmid":"38017546","id":"PMC_38017546","title":"hnRNPA2B1 promotes the occurrence and progression of hepatocellular carcinoma by downregulating PCK1 mRNA via a m6A RNA methylation manner.","date":"2023","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38017546","citation_count":20,"is_preprint":false},{"pmid":"24586865","id":"PMC_24586865","title":"Involvement of KLF11 in hepatic glucose metabolism in mice via suppressing of PEPCK-C expression.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24586865","citation_count":20,"is_preprint":false},{"pmid":"32522621","id":"PMC_32522621","title":"Hepatocellular carcinoma cell-derived extracellular vesicles encapsulated microRNA-584-5p facilitates angiogenesis through PCK1-mediated nuclear factor E2-related factor 2 signaling pathway.","date":"2020","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/32522621","citation_count":20,"is_preprint":false},{"pmid":"8985654","id":"PMC_8985654","title":"Indication for genetic linkage of the phosphoenolpyruvate carboxykinase (PCK1) gene region on chromosome 20q to non-insulin-dependent diabetes mellitus.","date":"1996","source":"Diabetes & metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/8985654","citation_count":20,"is_preprint":false},{"pmid":"30805369","id":"PMC_30805369","title":"Relationships of SLC2A4, RBP4, PCK1, and PI3K Gene Polymorphisms with Gestational Diabetes Mellitus in a Chinese Population.","date":"2019","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/30805369","citation_count":19,"is_preprint":false},{"pmid":"26792594","id":"PMC_26792594","title":"c.A2456C-substitution in Pck1 changes the enzyme kinetic and functional properties modifying fat distribution in pigs.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26792594","citation_count":19,"is_preprint":false},{"pmid":"27248905","id":"PMC_27248905","title":"Gentiopicroside and sweroside from Veratrilla baillonii Franch. induce phosphorylation of Akt and suppress Pck1 expression in hepatoma cells.","date":"2016","source":"Biochemistry and cell biology = Biochimie et biologie cellulaire","url":"https://pubmed.ncbi.nlm.nih.gov/27248905","citation_count":19,"is_preprint":false},{"pmid":"27785616","id":"PMC_27785616","title":"PEPCK-C reexpression in the liver counters neonatal hypoglycemia in Pck1 del/del mice, unmasking role in non-gluconeogenic tissues.","date":"2016","source":"Journal of physiology and biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27785616","citation_count":18,"is_preprint":false},{"pmid":"27170658","id":"PMC_27170658","title":"Chronic anemic hypoxemia increases plasma glucagon and hepatic PCK1 mRNA in late-gestation fetal sheep.","date":"2016","source":"American journal of physiology. Regulatory, integrative and comparative physiology","url":"https://pubmed.ncbi.nlm.nih.gov/27170658","citation_count":18,"is_preprint":false},{"pmid":"38085830","id":"PMC_38085830","title":"m 6 A-mediated gluconeogenic enzyme PCK1 upregulation protects against hepatic ischemia-reperfusion injury.","date":"2023","source":"Hepatology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/38085830","citation_count":17,"is_preprint":false},{"pmid":"16282543","id":"PMC_16282543","title":"Disparate associations of a functional promoter polymorphism in PCK1 with carotid wall ultrasound traits.","date":"2005","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/16282543","citation_count":17,"is_preprint":false},{"pmid":"37719384","id":"PMC_37719384","title":"Hepatic retinaldehyde deficiency is involved in diabetes deterioration by enhancing PCK1- and G6PC-mediated gluconeogenesis.","date":"2023","source":"Acta pharmaceutica Sinica. B","url":"https://pubmed.ncbi.nlm.nih.gov/37719384","citation_count":16,"is_preprint":false},{"pmid":"8432541","id":"PMC_8432541","title":"Human PCK1 encoding phosphoenolpyruvate carboxykinase is located on chromosome 20q13.2.","date":"1993","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8432541","citation_count":16,"is_preprint":false},{"pmid":"21152065","id":"PMC_21152065","title":"A putative Alzheimer's disease risk allele in PCK1 influences brain atrophy in multiple sclerosis.","date":"2010","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21152065","citation_count":16,"is_preprint":false},{"pmid":"35910380","id":"PMC_35910380","title":"Nintedanib Alleviates Experimental Colitis by Inhibiting CEBPB/PCK1 and CEBPB/EFNA1 Pathways.","date":"2022","source":"Frontiers in pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/35910380","citation_count":15,"is_preprint":false},{"pmid":"7781998","id":"PMC_7781998","title":"Thirteen genes (Cebpb, E2f1, Tcf4, Cyp24, Pck1, Acra4, Edn3, Kcnb1, Mc3r, Ntsr, Cd40, Plcg1 and Rcad) that probably lie in the distal imprinting region of mouse chromosome 2 are not monoallelically expressed.","date":"1995","source":"Genetical research","url":"https://pubmed.ncbi.nlm.nih.gov/7781998","citation_count":13,"is_preprint":false},{"pmid":"39433189","id":"PMC_39433189","title":"Porcine reproductive and respiratory syndrome virus activates lipid synthesis through a ROS-dependent AKT/PCK1/INSIG/SREBPs axis.","date":"2024","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/39433189","citation_count":13,"is_preprint":false},{"pmid":"32908218","id":"PMC_32908218","title":"Novel missense variants in PCK1 gene cause cytosolic PEPCK deficiency with growth failure from inadequate caloric intake.","date":"2020","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/32908218","citation_count":12,"is_preprint":false},{"pmid":"1492743","id":"PMC_1492743","title":"The genes coding for phosphoenolpyruvate carboxykinase-1 (PCK1) and neuronal nicotinic acetylcholine receptor alpha 4 subunit (CHRNA4) map to human chromosome 20, extending the known region of homology with mouse chromosome 2.","date":"1992","source":"Annals of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/1492743","citation_count":12,"is_preprint":false},{"pmid":"38651689","id":"PMC_38651689","title":"Inhibition of Pck1 in intestinal epithelial cells alleviates acute pancreatitis via modulating intestinal homeostasis.","date":"2024","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/38651689","citation_count":11,"is_preprint":false},{"pmid":"36840800","id":"PMC_36840800","title":"Role of phosphoenolpyruvate carboxykinase 1 (pck1) in mediating nutrient metabolism in zebrafish.","date":"2023","source":"Functional & integrative genomics","url":"https://pubmed.ncbi.nlm.nih.gov/36840800","citation_count":11,"is_preprint":false},{"pmid":"26725319","id":"PMC_26725319","title":"PCK1 is negatively regulated by bta-miR-26a, and a single-nucleotide polymorphism in the 3' untranslated region is involved in semen quality and longevity of Holstein bulls.","date":"2016","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/26725319","citation_count":11,"is_preprint":false},{"pmid":"16620271","id":"PMC_16620271","title":"Association of the promoter polymorphism -232C/G of the phosphoenolpyruvate carboxykinase gene (PCK1) with Type 2 diabetes mellitus.","date":"2006","source":"Diabetic medicine : a journal of the British Diabetic Association","url":"https://pubmed.ncbi.nlm.nih.gov/16620271","citation_count":11,"is_preprint":false},{"pmid":"38871968","id":"PMC_38871968","title":"PCK1-mediated glycogenolysis facilitates ROS clearance and chemotherapy resistance in cervical cancer stem cells.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/38871968","citation_count":10,"is_preprint":false},{"pmid":"34815524","id":"PMC_34815524","title":"PCK1 regulates neuroendocrine differentiation in a positive feedback loop of LIF/ZBTB46 signalling in castration-resistant prostate cancer.","date":"2021","source":"British journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/34815524","citation_count":10,"is_preprint":false},{"pmid":"18405640","id":"PMC_18405640","title":"[Glyceroneogenesis and PEPCK-C: pharmacological targets in type 2 diabetes].","date":"2008","source":"Medecine sciences : M/S","url":"https://pubmed.ncbi.nlm.nih.gov/18405640","citation_count":9,"is_preprint":false},{"pmid":"37062825","id":"PMC_37062825","title":"PCK1 activates oncogenic autophagy via down-regulation Serine phosphorylation of UBAP2L and antagonizes colorectal cancer growth.","date":"2023","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/37062825","citation_count":9,"is_preprint":false},{"pmid":"32181286","id":"PMC_32181286","title":"Transcriptomic changes associated with PCK1 overexpression in hepatocellular carcinoma cells detected by RNA-seq.","date":"2019","source":"Genes & diseases","url":"https://pubmed.ncbi.nlm.nih.gov/32181286","citation_count":9,"is_preprint":false},{"pmid":"36092708","id":"PMC_36092708","title":"NFYA promotes the anti-tumor effects of gluconeogenesis in hepatocellular carcinoma through the regulation of PCK1 expression.","date":"2022","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/36092708","citation_count":9,"is_preprint":false},{"pmid":"36455788","id":"PMC_36455788","title":"T3 and glucose increase expression of phosphoenolpyruvate carboxykinase (PCK1) leading to increased β-cell proliferation.","date":"2022","source":"Molecular metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/36455788","citation_count":9,"is_preprint":false},{"pmid":"39907589","id":"PMC_39907589","title":"Positively Charged Nanoplastics Destruct the Structure of the PCK1 Enzyme, Promote the Aerobic Gycolysis Pathway, and Induce Hepatic Tumor Risks.","date":"2025","source":"Environmental science & technology","url":"https://pubmed.ncbi.nlm.nih.gov/39907589","citation_count":9,"is_preprint":false},{"pmid":"20134411","id":"PMC_20134411","title":"Lack of association between PCK1 polymorphisms and obesity, physical activity, and fitness in European Youth Heart Study (EYHS).","date":"2010","source":"Obesity (Silver Spring, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/20134411","citation_count":8,"is_preprint":false},{"pmid":"16978381","id":"PMC_16978381","title":"Large-scale study of the -232C > G polymorphism of PCK1 in Type 2 diabetes.","date":"2006","source":"Diabetic medicine : a journal of the British Diabetic Association","url":"https://pubmed.ncbi.nlm.nih.gov/16978381","citation_count":8,"is_preprint":false},{"pmid":"32104690","id":"PMC_32104690","title":"PCK1 Deficiency Shortens the Replicative Lifespan of Saccharomyces cerevisiae through Upregulation of PFK1.","date":"2020","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/32104690","citation_count":8,"is_preprint":false},{"pmid":"19329064","id":"PMC_19329064","title":"Identification of conserved regulatory elements in mammalian promoter regions: a case study using the PCK1 promoter.","date":"2008","source":"Genomics, proteomics & bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/19329064","citation_count":8,"is_preprint":false},{"pmid":"37595871","id":"PMC_37595871","title":"SHP-1 phosphatase acts as a coactivator of PCK1 transcription to control gluconeogenesis.","date":"2023","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37595871","citation_count":7,"is_preprint":false},{"pmid":"33334880","id":"PMC_33334880","title":"Self-acetylation at the active site of phosphoenolpyruvate carboxykinase (PCK1) controls enzyme activity.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/33334880","citation_count":7,"is_preprint":false},{"pmid":"39463224","id":"PMC_39463224","title":"SIRT2 Alleviates Inflammatory Response, Apoptosis, and ECM Degradation in Osteoarthritic Chondrocytes by Stabilizing PCK1.","date":"2024","source":"Discovery medicine","url":"https://pubmed.ncbi.nlm.nih.gov/39463224","citation_count":7,"is_preprint":false},{"pmid":"25807515","id":"PMC_25807515","title":"Phosphoenolpyruvate carboxykinase 1 gene (Pck1) displays parallel evolution between Old World and New World fruit bats.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25807515","citation_count":7,"is_preprint":false},{"pmid":"39578429","id":"PMC_39578429","title":"PCK1 as a target for cancer therapy: from metabolic reprogramming to immune microenvironment remodeling.","date":"2024","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/39578429","citation_count":7,"is_preprint":false},{"pmid":"28198082","id":"PMC_28198082","title":"PCK1 expression is correlated with the plasma glucose level in the duck.","date":"2017","source":"Animal genetics","url":"https://pubmed.ncbi.nlm.nih.gov/28198082","citation_count":7,"is_preprint":false},{"pmid":"30805021","id":"PMC_30805021","title":"Role of PCK1 gene on oil tea-induced glucose homeostasis and type 2 diabetes: an animal experiment and a case-control study.","date":"2019","source":"Nutrition & metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/30805021","citation_count":7,"is_preprint":false},{"pmid":"40048154","id":"PMC_40048154","title":"PCK1 inhibits cGAS-STING activation by consumption of GTP to promote tumor immune evasion.","date":"2025","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40048154","citation_count":6,"is_preprint":false},{"pmid":"19070910","id":"PMC_19070910","title":"A Leu184Val polymorphism in PCK1 gene is associated with type 2 diabetes in Eastern Chinese population with BMI<23 kg/m2.","date":"2008","source":"Diabetes research and clinical practice","url":"https://pubmed.ncbi.nlm.nih.gov/19070910","citation_count":6,"is_preprint":false},{"pmid":"37637941","id":"PMC_37637941","title":"Emerging roles of cytosolic phosphoenolpyruvate kinase 1 (PCK1) in cancer.","date":"2023","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/37637941","citation_count":5,"is_preprint":false},{"pmid":"39341205","id":"PMC_39341205","title":"Small molecules targeting selective PCK1 and PGC-1α lysine acetylation cause anti-diabetic action through increased lactate oxidation.","date":"2024","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/39341205","citation_count":5,"is_preprint":false},{"pmid":"26280835","id":"PMC_26280835","title":"Association of PCK1 with Body Mass Index and Other Metabolic Features in Patients With Psychotropic Treatments.","date":"2015","source":"Journal of clinical psychopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/26280835","citation_count":5,"is_preprint":false},{"pmid":"31883179","id":"PMC_31883179","title":"Recognition of the gluconeogenic enzyme, Pck1, via the Gid4 E3 ligase: An in silico perspective.","date":"2019","source":"Journal of molecular recognition : JMR","url":"https://pubmed.ncbi.nlm.nih.gov/31883179","citation_count":5,"is_preprint":false},{"pmid":"24089092","id":"PMC_24089092","title":"Novel splice variants of the bovine PCK1 gene.","date":"2013","source":"Genetics and molecular research : GMR","url":"https://pubmed.ncbi.nlm.nih.gov/24089092","citation_count":5,"is_preprint":false},{"pmid":"33741163","id":"PMC_33741163","title":"Identification of promoter response elements that mediate propionate induction of bovine cytosolic phosphoenolpyruvate carboxykinase (PCK1) gene transcription.","date":"2021","source":"Journal of dairy science","url":"https://pubmed.ncbi.nlm.nih.gov/33741163","citation_count":5,"is_preprint":false},{"pmid":"34658252","id":"PMC_34658252","title":"Chronic AT1 blockade improves hyperglycemia by decreasing adipocyte inflammation and decreasing hepatic PCK1 and G6PC1 expression in obese rats.","date":"2021","source":"American journal of physiology. Endocrinology and metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/34658252","citation_count":5,"is_preprint":false},{"pmid":"31406102","id":"PMC_31406102","title":"Knockdown of the Sonic Hedgehog (SHH) Gene Inhibits Proliferation of Hep3B and SMMC-7721 Hepatocellular Carcinoma Cells via the PI3K/Akt/PCK1 Signaling Pathway.","date":"2019","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/31406102","citation_count":5,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":50867,"output_tokens":6951,"usd":0.128433},"stage2":{"model":"claude-opus-4-6","input_tokens":10631,"output_tokens":3922,"usd":0.226807},"total_usd":0.35524,"stage1_batch_id":"msgbatch_012ZiiCUBfB62NCTQd4Bk1Jo","stage2_batch_id":"msgbatch_01Xuqq9aUHoQouS7UVGZNStZ","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2020,\n      \"finding\": \"AKT phosphorylates PCK1 at Ser90, triggering its translocation from cytosol to the endoplasmic reticulum, where phosphorylated PCK1 acts as a GTP-dependent protein kinase to phosphorylate INSIG1 at Ser207 and INSIG2 at Ser151. This phosphorylation reduces sterol binding to INSIG1/2, disrupts INSIG–SCAP interaction, and releases the SCAP–SREBP complex to translocate to the Golgi, activating lipogenic gene transcription.\",\n      \"method\": \"In vitro kinase assays, site-directed mutagenesis, Co-IP, subcellular fractionation, ER translocation imaging, xenograft tumor models, patient HCC sample correlation\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution of kinase activity, mutagenesis of phosphosites, multiple orthogonal methods in single study, replicated in vivo\",\n      \"pmids\": [\"32322062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PCK1 acetylates itself at its active site (self-acetylation) using acetyl-CoA as a substrate, independently of p300. Site-directed acetylation of K244 inside the active site renders the enzyme inactive, producing a ~3-fold decrease in kcat without changing Km, revealing acetyl-CoA as a regulatory ligand that controls PCK1 gluconeogenic activity.\",\n      \"method\": \"Protein crystallography, mass spectrometry, isothermal titration calorimetry, saturation-transfer difference NMR, molecular docking, in vitro acetylation assay, site-directed mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstitution in vitro with structure, mutagenesis, and multiple biophysical methods in one study\",\n      \"pmids\": [\"33334880\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"SR18292 increases PCK1 lysine acetylation (K91 mimicked by K91Q mutant), which reverses PCK1 catalytic direction to favor anaplerotic OAA synthesis from PEP, thereby supplying OAA to the TCA cycle, increasing lactate and glucose oxidation, and suppressing gluconeogenesis. Liver-specific expression of PCK1 K91Q ameliorates hyperglycemia in obese mice.\",\n      \"method\": \"In vitro enzyme activity assays with acetylation-mimetic mutant (K91Q), metabolic flux analysis, liver-specific viral expression in mice, pharmacological treatment with SR18292\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — mutagenesis plus in vivo rescue, mechanistic confirmation of reverse catalytic reaction\",\n      \"pmids\": [\"39341205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Hypoxia induces JNK1/2-mediated phosphorylation of PCK1 at S151, which promotes physical interaction between PCK1 and cGAS. PCK1 associated with cGAS competitively consumes GTP (a shared substrate), thereby inhibiting GTP-dependent cGAS activation, suppressing STING-mediated immune cell recruitment, and promoting tumor immune evasion.\",\n      \"method\": \"Co-IP, in vitro GTP competition assays, phosphomimetic/phosphodeficient mutants, mouse tumor models with PCK1 blockade + anti-PD-1 combination, correlation in human breast cancer specimens\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — biochemical reconstitution of competition, mutagenesis, and in vivo tumor models with multiple orthogonal approaches\",\n      \"pmids\": [\"40048154\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PCK1 deficiency causes oxaloacetate accumulation and increased de novo UTP synthesis, elevating UDP-GlcNAc and global O-GlcNAcylation. Simultaneously, PCK1 loss inactivates the AMPK–GFAT1 axis, further promoting UDP-GlcNAc synthesis. Elevated O-GlcNAcylation of CHK2 at Thr378 counteracts CHK2 stability and dimer formation, increasing CHK2-dependent Rb phosphorylation and HCC cell proliferation.\",\n      \"method\": \"PCK1 knockout cells, metabolic flux analysis, AMPK pathway knockdown/rescue, O-GlcNAc mass spectrometry, site-specific mutagenesis of CHK2 T378, liver-specific Pck1 knockout mouse HCC model\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (metabolomics, genetic rescue, site mutagenesis, in vivo model) in a single study\",\n      \"pmids\": [\"33690219\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PCK1 depletion increases O-GlcNAcylation of KAT5 (lysine acetyltransferase 5), which suppresses KAT5 ubiquitination and stabilizes it. Stabilized O-GlcNAc-KAT5 epigenetically activates TWIST1 via histone H4 acetylation and upregulates MMP9/MMP14 via c-Myc acetylation, promoting EMT and HCC metastasis.\",\n      \"method\": \"Gain/loss-of-function studies, Co-IP, ubiquitination assays, ChIP, O-GlcNAc site mapping by MS, in vivo lung metastasis in liver-specific Pck1-KO mice\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal assays (Co-IP, ChIP, ubiquitination, in vivo metastasis) demonstrating mechanistic chain\",\n      \"pmids\": [\"34650217\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PCK1 fuels the serine synthesis pathway to generate S-adenosylmethionine (SAM). SAM serves as a methyl donor for SUV39H1-catalyzed H3K9me3 modification at the S100A11 promoter, suppressing this oncogene. PCK1 deficiency reduces SAM, lowers H3K9me3 at S100A11, and induces oncogenic PI3K/AKT signaling via S100A11–AKT1 interaction.\",\n      \"method\": \"Metabolomic tracing of serine/SAM pathway, ChIP for H3K9me3, Co-IP of S100A11–AKT1, SAM supplementation rescue, S100A11 KO in vivo, PCK1 KO/OE hepatoma models\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multi-omics plus genetic rescue in vitro and in vivo, mechanistically linking PCK1 to epigenetic control via SAM\",\n      \"pmids\": [\"37166978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CD8+ memory T cells upregulate PCK1 to drive gluconeogenesis-dependent glycogenesis. The resulting glycogen is channeled through glycogenolysis → glucose-6-phosphate → pentose phosphate pathway, generating NADPH that maintains reduced glutathione and limits ROS. Abrogation of the Pck1–glycogen–PPP axis impairs memory T-cell formation and maintenance.\",\n      \"method\": \"Pck1 KO and overexpression in T cells, metabolic flux analysis (glycogen, PPP intermediates, NADPH/GSH), mouse tumor immunotherapy models\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic KO with defined metabolic readouts, rescue experiments, and in vivo immunotherapy validation\",\n      \"pmids\": [\"29230018\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Forced PCK1 expression in glucose-deprived liver cancer cells induces TCA cataplerosis, leading to energy crisis and oxidative stress that causes apoptosis. This pro-apoptotic effect requires PCK1 catalytic activity, as catalytic-dead mutants fail to recapitulate it. Replenishing α-ketoglutarate or inhibiting ROS blocks the PCK1-induced cell death.\",\n      \"method\": \"PCK1 overexpression with catalytic mutant controls, TCA metabolite profiling, ROS measurement, α-ketoglutarate rescue, xenograft tumor models\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1/2 — catalytic-dead mutagenesis plus metabolic rescue establishes enzymatic mechanism\",\n      \"pmids\": [\"29335519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PCK1 upregulation in metastatic colorectal cancer cells drives pyrimidine nucleotide biosynthesis under hypoxia by supplying carbon via the gluconeogenic reaction, thereby supporting liver metastatic colonization and hypoxic growth.\",\n      \"method\": \"In vivo PDX selection for metastatic capacity, metabolomic profiling of pyrimidine intermediates, DHODH inhibition with leflunomide, PCK1 functional knockdown in metastasis models\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vivo selection model with metabolomics and pharmacological inhibition, but detailed biochemical mechanism of carbon channeling not fully reconstituted\",\n      \"pmids\": [\"31841108\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PCK1 deficiency inactivates AMPK, suppresses p27Kip1 expression, and stimulates the CDK/Rb/E2F pathway, accelerating G1→S transition and hepatoma cell proliferation under glucose starvation. Conversely, PCK1 overexpression reduces ATP, enhances AMPK phosphorylation, and induces G1 arrest; AMPK knockdown reverses the PCK1-induced arrest.\",\n      \"method\": \"Gain/loss-of-function with AMPK inhibitor/activator, flow cytometry cell-cycle analysis, Western blotting of CDK/Rb/E2F/AMPK/p27, xenograft models\",\n      \"journal\": \"Journal of experimental & clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic rescue (AMPK KD reversal) and pharmacological confirmation with defined phenotypic readout\",\n      \"pmids\": [\"30717766\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"SHP-1 phosphatase is recruited to the regulatory regions of the PCK1 gene, interacts with RNA polymerase II, and acts as a transcriptional coactivator for PCK1 expression. This recruitment is dependent on SHP-1's association with the transcription factor STAT5. Loss of SHP-1 or STAT5 reduces RNA Pol II at the PCK1 promoter, decreasing PCK1 mRNA and gluconeogenesis.\",\n      \"method\": \"Co-IP, ChIP-seq, luciferase reporter assay, gluconeogenesis assay, siRNA knockdown of SHP-1 and STAT5\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP-seq + Co-IP + functional gluconeogenesis rescue, single lab\",\n      \"pmids\": [\"37595871\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Insulin-induced Ser256 phosphorylation of FOXO1 triggers its translocation from nuclear speckles to the nuclear periphery, where FOXO1 forms a complex with EHMT2 histone methyltransferase. This complex induces repressive histone modifications at the PCK1 promoter and requires NUP98 for nuclear peripheral positioning, leading to transcriptional repression of PCK1.\",\n      \"method\": \"Live-cell imaging of endogenous phospho-FOXO1, Co-IP of FOXO1-EHMT2, ChIP of histone marks at PCK1 promoter, dominant-negative FOXO1 mutants, NUP98 interaction studies\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods establishing a new FOXO1 nuclear mechanism, single lab\",\n      \"pmids\": [\"25736587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The oncoprotein HBXIP suppresses PCK1 expression by: (1) upregulating miR-135a, which targets the 3′UTR of FOXO1 mRNA, and (2) activating PI3K/Akt to phosphorylate FOXO1, causing its nuclear export. Both mechanisms reduce FOXO1-driven PCK1 transcription. Overexpression of PCK1 abolishes HBXIP-promoted hepatoma growth in vitro and in vivo.\",\n      \"method\": \"miR-135a reporter assay (3′UTR luciferase), FOXO1 phosphorylation/localization by Western blot and IF, PCK1 rescue overexpression, xenograft models\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — mechanistic chain established with multiple assays but heavily reliant on overexpression/knockdown without site mutagenesis\",\n      \"pmids\": [\"27609066\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"METTL3-mediated N6-methyladenosine (m6A) modification of PCK1 mRNA is induced during hepatic ischemia-reperfusion (I/R) injury, promoting PCK1 mRNA export and increased PCK1 expression. Hepatic-specific knockout of METTL3 reduces m6A on PCK1 transcript, decreases PCK1 expression, and worsens I/R injury, while PCK1 overexpression is protective.\",\n      \"method\": \"MeRIP-seq, hepatocyte-specific METTL3 KO mice, I/R mouse models, PCK1 inhibitor and overexpression, mRNA export assays\",\n      \"journal\": \"Hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — MeRIP-seq mechanistically links METTL3 to PCK1 mRNA regulation in vivo, with genetic KO and functional rescue\",\n      \"pmids\": [\"38085830\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PCK1 loss in hepatoma cells increases global protein O-GlcNAcylation through two mechanisms: (1) oxaloacetate accumulation drives increased de novo UTP and UDP-GlcNAc synthesis, and (2) AMPK inactivation releases inhibitory GFAT1 phosphorylation. hnRNPA2B1 promotes HCC by binding PCK1 mRNA and reducing its m6A methylation, thereby decreasing PCK1 expression.\",\n      \"method\": \"RNA immunoprecipitation (RIP), MeRIP assay for m6A, CRISPR-Cas9 KO of hnRNPA2B1, RNA-seq, subcutaneous and orthotopic HCC mouse models\",\n      \"journal\": \"Journal of translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — RIP and MeRIP establish direct binding and m6A mechanism; in vivo rescue confirms functional dependence on PCK1\",\n      \"pmids\": [\"38017546\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Loss of PCK1 in myeloid cells (macrophage-specific Pck1 deletion) reduces 13C labeling of TCA intermediates (citrate, malate) from [U-13C]glucose, increases 13C-lactate labeling, and elevates ROS, shifting macrophages toward a pro-inflammatory M1 phenotype with increased TNFα, IL-1β, and IL-6 expression.\",\n      \"method\": \"Myeloid-specific Pck1 KO mice, stable isotopomer MS with [U-13C]glucose in BMDM, ROS measurement, cytokine ELISA\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — isotope tracing in genetic KO macrophages directly links PCK1 to macrophage metabolic reprogramming and inflammatory phenotype\",\n      \"pmids\": [\"29317502\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"A single amino acid substitution Met139Leu (c.A2456C SNP) in pig PCK1 reduces kcat in the glyceroneogenic direction, enhances kcat in the anaplerotic direction, and reduces ability of the enzyme to be acetylated, increasing its susceptibility to ubiquitin-proteasome degradation. The substitution results in ~30% lower glucose and ~9% lower lipid production in cell cultures.\",\n      \"method\": \"Recombinant enzyme kinetic assays (kcat, Km in both directions), acetylation assay, cell culture metabolic flux measurement, pig phenotype association\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzymatic characterization with defined mutant, plus post-translational modification consequence, single lab\",\n      \"pmids\": [\"26792594\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Kidney-specific PCK1 deletion in mice causes hyperchloremic metabolic acidosis with reduced ammoniagenesis, glycosuria, lactaturia, and decreased ATP generation. PCK1 is required for proximal tubule energy production and acid-base control; its loss increases tubular injury during acidosis, while overexpression protects against proteinuric renal disease.\",\n      \"method\": \"Kidney-specific PCK1 KO and knockin (PAX8 promoter), metabolic phenotyping (creatinine clearance, albuminuria, glycosuria), ATP measurement, acid-base challenge experiments\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — tissue-specific genetic models with defined functional readouts (acid-base, energy, injury), single lab\",\n      \"pmids\": [\"37102687\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"PCK1 loss in HCC deficiency activates the RhoA/PI3K/AKT pathway by increasing intracellular GTP levels, stimulates paracrine secretion of PDGF-AA, and promotes hepatic stellate cell activation and liver fibrosis. Treatment with RhoA and AKT inhibitors or silencing of RhoA/AKT1 alleviates MAFLD progression in PCK1-deficient mice.\",\n      \"method\": \"Liver-specific Pck1 KO mice, intracellular GTP measurement, PDGF-AA ELISA, hepatic stellate cell co-culture, RhoA/AKT inhibitor treatment, AAV-mediated PCK1 rescue\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — GTP measurement mechanistically links PCK1 substrate consumption to RhoA/PI3K pathway; genetic and pharmacological rescue in vivo\",\n      \"pmids\": [\"36918564\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PCK1 enhances PYGL phosphorylation in cervical cancer stem cells, promoting glycogen breakdown and channeling glucose-6-phosphate into the pentose phosphate pathway, increasing NADPH production and ROS clearance, thereby promoting chemoresistance.\",\n      \"method\": \"siRNA knockdown of PCK1, PYGL, GYS1 in HCC94/CaSki cells, glycogen measurement, LC-MS of PPP intermediates, NADPH/NADP+ ratio, NSG mouse tumor growth assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct biochemical measurement of PPP/NADPH with genetic perturbations and in vivo tumor model\",\n      \"pmids\": [\"38871968\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"PRRSV infection activates AKT, which in turn activates PCK1. Activated PCK1 phosphorylates INSIG proteins (INSIGs), causing their degradation and releasing SCAP–SREBP complexes to the nucleus to activate lipid biosynthesis; ROS produced by PRRSV are required for AKT activation in this axis.\",\n      \"method\": \"Pharmacological inhibitors of AKT and PCK1, INSIG protein level monitoring by Western blot, SREBP nuclear translocation assay, ROS scavenger treatment, MARC-145 cell infection model\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — mechanistic evidence relies primarily on pharmacological inhibitors and Western blot in cell culture, limited genetic validation\",\n      \"pmids\": [\"39433189\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Rev-erbα directly binds a RevRE site at −325 to −320 bp in the PCK1 promoter to trans-repress PCK1 transcription, reducing hepatic Pck1 mRNA and protein levels and lowering fasting plasma glucose in diabetic mice.\",\n      \"method\": \"Luciferase reporter assay with promoter deletions, gel-shift (EMSA) with RevRE site probe, ChIP assay in hepatoma cells, SR9009 (Rev-erbα agonist) treatment in WT and STZ diabetic mice\",\n      \"journal\": \"Pharmacological research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — EMSA + ChIP + luciferase + in vivo pharmacology establish direct promoter binding and functional consequence\",\n      \"pmids\": [\"30639375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PCK1 antagonizes CRC cell growth by inactivating UBAP2L phosphorylation at serine 454, thereby enhancing autophagy flux and suppressing tumor proliferation in vitro and in vivo.\",\n      \"method\": \"Overexpression and knockdown of PCK1, phosphoproteomic identification of UBAP2L pS454, autophagy flux assay (LC3, p62), xenograft models\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — phosphosite identified but kinase-substrate relationship not biochemically reconstituted; single lab\",\n      \"pmids\": [\"37062825\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The −232C>G promoter SNP of human PCK1 lies within a cis-acting element required for basal and cAMP-mediated transcription. The −232G allele in luciferase reporter assays shows increased basal expression and loss of insulin-mediated downregulation, establishing that this element is required for insulin suppression of PCK1 transcription.\",\n      \"method\": \"Luciferase reporter gene assay in three cell lines with −232C vs −232G constructs, insulin suppression experiments\",\n      \"journal\": \"The Journal of clinical endocrinology and metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional reporter assay in multiple cell lines with specific promoter element directly tested\",\n      \"pmids\": [\"14764811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"PCK1 overexpression in proximal tubules suppresses HK2 upregulation (rate-limiting enzyme of glycolysis), thereby blocking excess glycolysis. PCK1 overexpression preserves mitoribosomal function and prevents tubular fibrosis in diabetic nephropathy; conversely, PCK1 CKO mice develop mitoribosomal defects and renal fibrosis.\",\n      \"method\": \"PT-specific Pck1 TG and CKO mice, STZ diabetic model, Western blot for mitoribosomes and HK2, type IV collagen deposition, albuminuria measurement\",\n      \"journal\": \"Journal of the American Society of Nephrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — bidirectional genetic models (TG and CKO) in disease model with defined molecular and histological readouts\",\n      \"pmids\": [\"37199399\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PCK1 (cytosolic PEPCK) is a multifunctional enzyme that catalyzes the GTP-dependent conversion of oxaloacetate to phosphoenolpyruvate for gluconeogenesis, but is also regulated post-translationally: AKT-mediated Ser90 phosphorylation redirects PCK1 to the ER where it acts as a GTP-dependent protein kinase phosphorylating INSIG1/2 to activate SREBP-driven lipogenesis; acetyl-CoA binds the active site and drives self-acetylation (notably at K244/K91) to inhibit gluconeogenic activity and favor anaplerotic OAA synthesis; JNK1/2-mediated S151 phosphorylation promotes PCK1–cGAS interaction that competitively consumes GTP to suppress innate immune signaling; and through its enzymatic activity PCK1 controls TCA cataplerosis, glycogenesis/PPP flux for NADPH and ROS homeostasis, SAM production for H3K9 methylation, and O-GlcNAcylation levels, collectively regulating metabolism, epigenetics, cell-cycle progression via AMPK/p27, and immune evasion in cancer and normal physiology.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"PCK1 (cytosolic phosphoenolpyruvate carboxykinase) is a GTP-dependent enzyme whose canonical activity—conversion of oxaloacetate to phosphoenolpyruvate—positions it as a master regulator of carbon flux through gluconeogenesis, TCA cataplerosis, glycogenesis, the pentose phosphate pathway, and one-carbon/SAM metabolism, with consequences for NADPH/ROS homeostasis, epigenetic H3K9 trimethylation, O-GlcNAcylation, and cell-cycle control via AMPK/p27 [PMID:29230018, PMID:33690219, PMID:37166978, PMID:30717766]. Beyond its metabolic activity, AKT-mediated Ser90 phosphorylation redirects PCK1 to the endoplasmic reticulum where it functions as a protein kinase, phosphorylating INSIG1/2 to release SCAP–SREBP and activate lipogenesis [PMID:32322062]. Acetyl-CoA binding at the active site drives PCK1 self-acetylation (K244, K91), which inactivates the gluconeogenic reaction and can reverse catalytic directionality to favor anaplerotic oxaloacetate synthesis [PMID:33334880, PMID:39341205]. JNK1/2-mediated S151 phosphorylation promotes PCK1–cGAS interaction and competitive GTP consumption, suppressing cGAS–STING innate immune signaling and enabling tumor immune evasion [PMID:40048154].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Identification of a cis-regulatory element at −232 in the PCK1 promoter established that a single nucleotide variant governs basal transcription and insulin-mediated repression, providing the first functional dissection of how insulin silences PCK1 at the promoter level.\",\n      \"evidence\": \"Luciferase reporter assays with −232C vs −232G constructs in multiple cell lines with insulin treatment\",\n      \"pmids\": [\"14764811\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Chromatin context of the −232 element not examined\", \"Trans-acting factor binding this element not identified\", \"No in vivo confirmation of allele-specific expression\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Discovery that insulin-phosphorylated FOXO1 relocates to the nuclear periphery and recruits the histone methyltransferase EHMT2, establishing repressive chromatin at the PCK1 promoter, revealed a spatial–epigenetic mechanism of PCK1 transcriptional silencing beyond simple FOXO1 nuclear exclusion.\",\n      \"evidence\": \"Live-cell imaging of endogenous phospho-FOXO1, Co-IP of FOXO1–EHMT2, ChIP of histone marks at PCK1 promoter, NUP98 interaction studies\",\n      \"pmids\": [\"25736587\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Not independently confirmed by a second lab\", \"Contribution relative to canonical FOXO1 nuclear export not quantified\", \"Whether this mechanism operates genome-wide or is PCK1-specific is unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstration that CD8+ memory T cells upregulate PCK1 to fuel a gluconeogenesis→glycogenesis→PPP axis for NADPH and ROS control established a non-hepatic, immunological role for PCK1 and linked its catalytic activity to adaptive immune memory.\",\n      \"evidence\": \"T-cell-specific Pck1 KO and overexpression, 13C metabolic flux analysis, NADPH/GSH measurement, mouse tumor immunotherapy models\",\n      \"pmids\": [\"29230018\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PCK1 is required in all memory T-cell subsets is unknown\", \"Upstream signals inducing PCK1 in memory T cells not fully defined\", \"Relative contribution of glycogen vs direct PPP flux not dissected\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showing that forced PCK1 expression in glucose-deprived hepatoma cells causes lethal TCA cataplerosis, energy crisis, and ROS-mediated apoptosis—dependent on catalytic activity—established PCK1 as a context-dependent tumor suppressor whose metabolic drain is cytotoxic.\",\n      \"evidence\": \"PCK1 overexpression with catalytic-dead mutant controls, TCA metabolite profiling, α-ketoglutarate rescue, xenograft models\",\n      \"pmids\": [\"29335519\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Threshold of cataplerosis required for apoptosis not quantified\", \"Relevance to non-hepatic tumors not tested in this study\", \"Downstream ROS sensors mediating apoptosis not identified\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linking PCK1 loss to AMPK inactivation, p27 suppression, and accelerated G1→S transition provided a direct mechanism by which metabolic sensing through PCK1 controls cell-cycle progression in hepatoma cells.\",\n      \"evidence\": \"PCK1 gain/loss-of-function with AMPK inhibitor/activator, flow cytometry, CDK/Rb/E2F pathway analysis, xenograft models\",\n      \"pmids\": [\"30717766\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the ATP drop is the sole AMPK-activating signal is unresolved\", \"Contribution of other AMPK substrates beyond p27 not tested\", \"Single cancer type studied\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identification of PCK1 upregulation in metastatic colorectal cancer cells driving pyrimidine nucleotide biosynthesis under hypoxia revealed that the gluconeogenic reaction can supply carbon for nucleotide synthesis in non-hepatic tumors, expanding PCK1's oncogenic role.\",\n      \"evidence\": \"PDX in vivo selection, metabolomic profiling of pyrimidine intermediates, DHODH inhibition, PCK1 knockdown in metastasis models\",\n      \"pmids\": [\"31841108\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Exact carbon routing from PEP to pyrimidines not traced isotopically at single-step resolution\", \"Whether this is specific to liver metastases or generalizable is unclear\", \"No structural or enzymatic reconstitution\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"The landmark finding that AKT phosphorylates PCK1 at Ser90, causing ER translocation and a moonlighting protein kinase activity toward INSIG1/2, fundamentally redefined PCK1 as a signal-regulated enzyme with dual catalytic identities—metabolic and signaling.\",\n      \"evidence\": \"In vitro kinase assays, site-directed mutagenesis, Co-IP, subcellular fractionation, ER translocation imaging, xenograft and human HCC correlation\",\n      \"pmids\": [\"32322062\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full substrate scope of PCK1 kinase activity beyond INSIGs not surveyed\", \"Structural basis for GTP-dependent kinase mechanism not resolved\", \"Whether kinase activity occurs in non-tumor tissues is unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Crystal-structure-guided discovery that acetyl-CoA binds the PCK1 active site and drives self-acetylation at K244, inactivating the enzyme, established a direct feedback mechanism by which acetyl-CoA availability controls gluconeogenic flux independently of acetyltransferases.\",\n      \"evidence\": \"X-ray crystallography, mass spectrometry, ITC, STD-NMR, in vitro acetylation assay, site-directed mutagenesis\",\n      \"pmids\": [\"33334880\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological acetyl-CoA concentrations at which self-acetylation becomes rate-limiting not defined\", \"Whether deacetylases counteract this in real time in vivo is unclear\", \"Impact on protein kinase activity not tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Comprehensive metabolic tracing showed PCK1 loss causes OAA accumulation, increased UDP-GlcNAc, and global O-GlcNAcylation—including CHK2 at T378, which destabilizes CHK2 and drives Rb phosphorylation and proliferation—establishing a PCK1→O-GlcNAc→cell-cycle axis in HCC.\",\n      \"evidence\": \"PCK1 KO cells, metabolic flux analysis, AMPK–GFAT1 epistasis, O-GlcNAc MS, CHK2 T378 mutagenesis, liver-specific Pck1 KO HCC mice\",\n      \"pmids\": [\"33690219\", \"34650217\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Complete O-GlcNAc proteome altered by PCK1 loss not catalogued\", \"Whether O-GlcNAcylation changes are reversible upon PCK1 restoration in established tumors is unknown\", \"Relative contribution of OAA-driven vs AMPK-driven UDP-GlcNAc increase not quantified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Discovery that PCK1 fuels serine/SAM biosynthesis and thereby sustains SUV39H1-catalyzed H3K9me3 at the S100A11 promoter linked PCK1 catalytic activity to epigenetic gene silencing and identified a metabolic–epigenetic tumor-suppressive circuit.\",\n      \"evidence\": \"Metabolomic tracing of serine/SAM, ChIP for H3K9me3, Co-IP of S100A11–AKT1, SAM rescue, S100A11 KO in vivo, PCK1 KO/OE hepatoma models\",\n      \"pmids\": [\"37166978\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genome-wide H3K9me3 changes upon PCK1 loss not reported\", \"Whether other SAM-consuming methyltransferases are similarly affected is unclear\", \"Direct isotope tracing from PEP to SAM not shown at single-step resolution\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Kidney-specific PCK1 deletion caused hyperchloremic metabolic acidosis, reduced ammoniagenesis, and impaired tubular ATP production, establishing PCK1 as essential for renal acid-base homeostasis and energy metabolism beyond its hepatic role.\",\n      \"evidence\": \"Kidney-specific PCK1 KO and knockin mice, metabolic phenotyping, ATP measurement, acid-base challenge\",\n      \"pmids\": [\"37102687\", \"37199399\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether PCK1 compensates for PCK2 loss in kidney is not tested\", \"Mechanism linking PCK1 to ammoniagenesis not fully delineated\", \"Long-term renal outcomes of PCK1 overexpression not characterized\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showing that K91 acetylation reverses PCK1's catalytic direction to favor anaplerotic OAA synthesis from PEP, and that liver-specific K91Q expression ameliorates hyperglycemia in obese mice, revealed a post-translational switch that repurposes PCK1 from catabolic to anabolic function in vivo.\",\n      \"evidence\": \"In vitro enzyme activity with K91Q mimetic, metabolic flux analysis, liver-specific viral expression in obese mice, SR18292 pharmacology\",\n      \"pmids\": [\"39341205\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether K91 acetylation and K244 acetylation cooperate or are mutually exclusive is unresolved\", \"Deacetylase(s) responsible for reversing K91 acetylation in vivo not identified\", \"Effect of reverse reaction on non-hepatic tissues not explored\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identification of JNK1/2-mediated S151 phosphorylation driving PCK1–cGAS association and competitive GTP consumption revealed a non-enzymatic, immune-suppressive function of PCK1 that enables tumor immune evasion via cGAS–STING inhibition.\",\n      \"evidence\": \"Co-IP, in vitro GTP competition assays, phosphomimetic/phosphodeficient mutants, mouse tumor models with anti-PD-1, human breast cancer specimen correlation\",\n      \"pmids\": [\"40048154\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PCK1–cGAS interaction occurs in non-tumor immune contexts is unknown\", \"Structural basis of the PCK1–cGAS interface not determined\", \"Relative contribution of GTP depletion vs direct cGAS inhibition not fully separated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Major open questions include the full substrate scope of PCK1's protein kinase activity, the structural basis for its dual catalytic (metabolic and kinase) functions, how the multiple post-translational modifications (Ser90, S151, K91, K244 phosphorylation/acetylation) are hierarchically coordinated in vivo, and whether PCK1's immune-modulatory roles extend beyond the tumor microenvironment.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal structure of phospho-Ser90 ER-localized PCK1 in kinase conformation\", \"Integrated PTM code governing catalytic direction not mapped\", \"Non-cancer immune roles of PCK1 largely unexplored\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 3, 21]},\n      {\"term_id\": \"GO:0016853\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0016829\", \"supporting_discovery_ids\": [2, 8]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 8]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0, 21]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 2, 7, 8, 9, 17, 18]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4, 10, 19, 21]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3, 7, 16]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [6]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4, 10]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"INSIG1\",\n      \"INSIG2\",\n      \"AKT1\",\n      \"cGAS\",\n      \"CHK2\",\n      \"PYGL\",\n      \"UBAP2L\",\n      \"SCAP\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}