{"gene":"ZDHHC4","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2023,"finding":"CPT1A recruits the ER-localized ZDHHC4 to catalyze MAVS palmitoylation at Cys79, which promotes MAVS stabilization and activation by inhibiting K48-linked ubiquitination while facilitating K63-linked ubiquitination, thereby sustaining IFN-I response.","method":"Mechanistic cell-based assays, co-immunoprecipitation, palmitoylation assays, ubiquitination analysis, site-specific mutagenesis","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, site-specific mutagenesis (Cys79), multiple orthogonal readouts (palmitoylation, K48/K63 ubiquitination, IFN-I response), functionally validated in vivo","pmids":["38016475"],"is_preprint":false},{"year":2021,"finding":"ZDHHC4 physically associates with SARS-CoV-2 S protein and its overexpression promotes palmitoylation of the S protein cytoplasmic tail, which is required for S-mediated syncytia formation and pseudovirus entry but not plasma membrane targeting.","method":"Co-immunoprecipitation, palmitoylation assay, overexpression screen with multiple ZDHHC family members, pseudovirus entry assay, syncytia formation assay","journal":"Journal of medical virology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with functional readouts, single lab, multiple orthogonal assays but no site-specific mutagenesis for ZDHHC4 specifically","pmids":["34528721"],"is_preprint":false},{"year":2021,"finding":"ZDHHC4 palmitoylates KAI1 (CD82), which is required for KAI1 localization to the membrane surface; membrane-localized KAI1 then induces LIF via the Src/p53 pathway to suppress angiogenesis.","method":"KAI1 knockout mice, in vitro palmitoylation assay, in vivo angiogenesis models, mechanistic pathway analysis","journal":"Journal of hematology & oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional KO model with defined cellular phenotype, palmitoylation linked to localization and pathway activation, single lab","pmids":["34530889"],"is_preprint":false},{"year":2015,"finding":"ZDHHC4 physically interacts with the D2 dopamine receptor (D2R) and affects its palmitoylation status; palmitoylation at C443 is required for D2R plasma membrane expression and protein stability.","method":"Membrane yeast two-hybrid (MYTH) screen, co-immunoprecipitation, bioorthogonal click chemistry palmitoylation assay, site-directed mutagenesis","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MYTH screen confirmed by Co-IP, palmitoylation assay with mutagenesis, single lab","pmids":["26535572"],"is_preprint":false},{"year":2022,"finding":"ZDHHC4 palmitoylates GSK3β at Cys14, which increases GSK3β activity (decreasing p-Ser9, increasing p-Tyr216) and activates the EZH2-STAT3 signaling axis to promote GBM stem cell self-renewal and temozolomide resistance.","method":"Palmitoylation assay, site-directed mutagenesis (Cys14), knockdown/overexpression, phosphorylation analysis, STAT3/EZH2 pathway readouts in GBM stem cells","journal":"Oncogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — site-specific mutagenesis combined with functional pathway readouts, single lab, multiple orthogonal methods","pmids":["35606353"],"is_preprint":false},{"year":2022,"finding":"ZDHHC4 (together with ZDHHC8) catalyzes S-palmitoylation of NFATC4, which is required for NFATC4 trafficking from the cytoplasm to the nucleus; in Ppt1-/- mice reduced ZDHHC4/ZDHHC8 levels lower nuclear palmitoylated NFATC4, inhibiting IP3R1 expression and dysregulating lysosomal Ca2+ homeostasis.","method":"Identification of ZDHHC4 and ZDHHC8 as NFATC4 palmitoyl acyltransferases, subcellular fractionation, palmitoylation assay in Ppt1-/- mouse model, IP3R1 overexpression rescue experiment","journal":"Journal of inherited metabolic disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — palmitoylation assay linked to subcellular trafficking and downstream functional consequence, genetic model with rescue, single lab","pmids":["35150145"],"is_preprint":false},{"year":2025,"finding":"FoxO1 transcriptionally upregulates ZDHHC4 expression in the diabetic heart; ZDHHC4 S-acylates CD36, promoting its sarcolemmal localization; genetic silencing of ZDHHC4 decreases CD36 S-acylation and sarcolemmal content, reducing fatty acid oxidation and triglyceride storage and improving cardiac function.","method":"ChIP-seq, ChIP-qPCR, luciferase reporter assay, siRNA/shRNA silencing, cardiomyocyte-specific FoxO1 knockout mice, pharmacological FoxO1 inhibition, S-acylation assay across multiple species (rat, db/db mouse, pig, human iPSC-CMs)","journal":"Circulation research","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (ChIP-seq, luciferase, genetic KO, pharmacological inhibition), cross-species validation, direct mechanistic chain from FoxO1 to ZDHHC4 to CD36 palmitoylation to functional metabolic phenotype","pmids":["40357580"],"is_preprint":false},{"year":2024,"finding":"ZDHHC4 physically interacts with TRPV1 and catalyzes S-palmitoylation at C157, C362, C390, and C715, promoting TRPV1 degradation via the lysosome pathway; this modification is counterbalanced by depalmitoylase APT1, and the net palmitoylation state regulates inflammatory pain relief.","method":"Co-immunoprecipitation, site-directed mutagenesis of TRPV1 cysteines, palmitoylation assay, lysosomal degradation assay, electrophysiology, in vivo pain behavior","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — site-specific mutagenesis of four palmitoylation sites, electrophysiology, in vivo evidence, and identification of the opposing depalmitoylase, multiple orthogonal methods in one study","pmids":["39528731"],"is_preprint":false},{"year":2024,"finding":"ATF3 in macrophages increases ZDHHC4/5-mediated CD36 palmitoylation at C3, C7, C464, and C466, promoting CD36 plasma membrane localization; this contributes to regulation of fatty acid oxidation and protection against MASH.","method":"Overexpression/ablation of Atf3 in macrophages, palmitoylation assay with site identification, subcellular fractionation, in vivo MASH mouse model","journal":"Science advances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — site-specific palmitoylation with functional localization readout, but ZDHHC4 and ZDHHC5 not individually resolved; single lab","pmids":["39047111"],"is_preprint":false},{"year":2026,"finding":"ZDHHC4 palmitoylates ZEB-2 at C478 (identified by mass spectrometry); ZEB-2 interacts with ZDHHC4 through its N-terminal sequences; palmitoylation promotes ZEB-2 deubiquitination and protein stability, which is required for EMT in melanoma cells.","method":"Mass spectrometry substrate identification, Co-immunoprecipitation, site-directed mutagenesis (C478), knockdown functional assays (proliferation, migration, invasion), ubiquitination assay","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS substrate ID confirmed by Co-IP and mutagenesis, functional knockdown with defined phenotype, single lab","pmids":["41603995"],"is_preprint":false},{"year":2026,"finding":"LCP (lactucopicrin) directly binds to His72 of ZDHHC4, boosting its enzymatic activity; activated ZDHHC4 catalyzes palmitoylation of CCDC50, facilitating selective autophagic degradation of MAP2K4/MKK4, suppressing MAPK/JNK signaling and chondrocyte senescence in osteoarthritis.","method":"DARTS (drug affinity responsive target stability) structural binding assay, palmitoylation assay, autophagy flux assay, MAPK/JNK signaling readouts, in vivo OA mouse model","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding to ZDHHC4 His72 identified, palmitoylation linked to autophagy and signaling cascade, in vivo validation, single lab","pmids":["41566717"],"is_preprint":false},{"year":2025,"finding":"EVA1A deficiency transcriptionally enhances ZDHHC4/5 expression while repressing the depalmitoylase APT1, resulting in increased CD36 palmitoylation and plasma membrane localization; this promotes fatty acid uptake and impairs β-oxidation in hepatocytes.","method":"Hepatocyte-specific knockout of Eva1a, overexpression rescue, palmitoylation assay, subcellular fractionation, transcriptional analysis of ZDHHC4/5 and APT1","journal":"Research (Washington, D.C.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO with functional metabolic phenotype and palmitoylation readout, but ZDHHC4 and ZDHHC5 not individually resolved; single lab","pmids":["41306774"],"is_preprint":false}],"current_model":"ZDHHC4 is an ER/membrane-localized palmitoyl acyltransferase that catalyzes S-palmitoylation of diverse substrates—including MAVS (Cys79), TRPV1 (C157/C362/C390/C715), GSK3β (Cys14), ZEB-2 (C478), NFATC4, CD36, KAI1, D2R, and CCDC50—to regulate their stability, subcellular trafficking, and signaling activity in contexts ranging from innate immune activation and pain modulation to metabolic regulation, cancer stemness, and EMT, with its own expression controlled transcriptionally by FoxO1 in metabolic disease."},"narrative":{"mechanistic_narrative":"ZDHHC4 is an ER/membrane-localized palmitoyl acyltransferase that catalyzes S-palmitoylation of a structurally diverse set of substrates to control their stability, subcellular trafficking, and downstream signaling across innate immunity, pain, metabolism, and cancer [PMID:38016475, PMID:39528731, PMID:40357580]. A recurrent mechanistic theme is that ZDHHC4-mediated palmitoylation governs substrate fate through two routes: directing membrane or nuclear localization, and modulating ubiquitin-dependent stability. In innate immunity, ZDHHC4 is recruited by CPT1A to palmitoylate MAVS at Cys79, blocking K48-linked ubiquitination while favoring K63-linked ubiquitination to stabilize MAVS and sustain the type I interferon response [PMID:38016475]; an analogous stabilization mechanism operates on ZEB-2 (palmitoylation at C478 promoting deubiquitination) to drive EMT in melanoma [PMID:41603995]. ZDHHC4 also controls trafficking-dependent activity: it palmitoylates KAI1/CD82 and the D2 dopamine receptor to direct their plasma-membrane localization and stability [PMID:34530889, PMID:26535572], drives nuclear translocation of NFATC4 [PMID:35150145], and—in metabolic tissues—S-acylates CD36 to promote its sarcolemmal/plasma-membrane localization, fatty-acid uptake and oxidation, with ZDHHC4 expression itself transcriptionally upregulated by FoxO1 in the diabetic heart [PMID:40357580, PMID:39047111, PMID:41306774]. Conversely, ZDHHC4 palmitoylation of TRPV1 at C157/C362/C390/C715 targets the channel for lysosomal degradation, a modification opposed by the depalmitoylase APT1 and tuning inflammatory pain [PMID:39528731]. Through palmitoylation of GSK3β at Cys14 it activates the EZH2-STAT3 axis in glioblastoma stem cells [PMID:35606353], and palmitoylation of CCDC50 links it to selective autophagy of MAP2K4 and suppression of MAPK/JNK signaling [PMID:41566717]. Its enzymatic activity is regulated at the residue level, being boosted by the small molecule lactucopicrin binding His72 [PMID:41566717].","teleology":[{"year":2015,"claim":"Established ZDHHC4 as a bona fide acyltransferase acting on a membrane receptor, addressing whether it physically engages and modifies substrates to control their surface expression.","evidence":"Membrane yeast two-hybrid screen, Co-IP and click-chemistry palmitoylation assay with D2R cysteine mutagenesis","pmids":["26535572"],"confidence":"Medium","gaps":["Single lab without reciprocal in vitro reconstitution of ZDHHC4 enzymatic activity","Did not define ZDHHC4 catalytic residues","Trafficking mechanism downstream of palmitoylation not resolved"]},{"year":2021,"claim":"Extended ZDHHC4 substrate repertoire to a tetraspanin and a viral glycoprotein, showing palmitoylation controls membrane localization and downstream signaling versus viral fusion.","evidence":"KAI1 knockout mice with palmitoylation and angiogenesis assays; Co-IP and pseudovirus/syncytia assays for SARS-CoV-2 S protein","pmids":["34530889","34528721"],"confidence":"Medium","gaps":["No ZDHHC4-specific site mutagenesis for the S protein","Direct catalytic role not separated from family redundancy in the viral study"]},{"year":2022,"claim":"Showed ZDHHC4 palmitoylation can both activate a kinase and license a transcription factor's nuclear entry, broadening its role to oncogenic and Ca2+-homeostasis signaling.","evidence":"Site-directed mutagenesis (GSK3β Cys14) with EZH2-STAT3 readouts in GBM stem cells; NFATC4 palmitoylation and fractionation in Ppt1-/- mice with IP3R1 rescue","pmids":["35606353","35150145"],"confidence":"Medium","gaps":["NFATC4 modification shared with ZDHHC8 and not individually resolved","Direct enzyme-substrate kinetics not established"]},{"year":2023,"claim":"Defined the highest-confidence mechanism: CPT1A-recruited ZDHHC4 palmitoylates MAVS at Cys79 to redirect ubiquitin signals (suppressing K48, favoring K63) and sustain interferon responses.","evidence":"Reciprocal Co-IP, Cys79 mutagenesis, palmitoylation and K48/K63 ubiquitination assays, in vivo IFN-I readouts","pmids":["38016475"],"confidence":"High","gaps":["Structural basis of CPT1A-ZDHHC4 recruitment not solved","Whether MAVS palmitoylation is reversed by a specific depalmitoylase unaddressed"]},{"year":2024,"claim":"Connected ZDHHC4 to substrate degradation rather than stabilization, and to physiological metabolism, establishing context-dependent outcomes of palmitoylation.","evidence":"TRPV1 four-site mutagenesis with lysosomal degradation, electrophysiology and pain behavior plus APT1 opposition; ATF3-driven CD36 palmitoylation with fractionation in a MASH model","pmids":["39528731","39047111"],"confidence":"High","gaps":["CD36 study did not individually resolve ZDHHC4 from ZDHHC5","How substrate identity dictates lysosomal versus stabilizing fate is unexplained"]},{"year":2025,"claim":"Placed ZDHHC4 within transcriptional regulatory circuits controlling cardiac and hepatic lipid handling via CD36 palmitoylation.","evidence":"FoxO1 ChIP-seq/luciferase and cardiomyocyte FoxO1 KO with cross-species S-acylation assays; hepatocyte Eva1a KO with ZDHHC4/5 and APT1 transcriptional analysis","pmids":["40357580","41306774"],"confidence":"High","gaps":["EVA1A study did not resolve ZDHHC4 from ZDHHC5","Direct ZDHHC4 enzymatic specificity for CD36 cysteines not isolated from family members"]},{"year":2026,"claim":"Identified a stabilizing substrate (ZEB-2) and a pharmacological activation handle (His72), advancing ZDHHC4 as a druggable enzyme with defined regulatory residues.","evidence":"Mass spectrometry, Co-IP and C478 mutagenesis for ZEB-2; DARTS binding to His72 by lactucopicrin with CCDC50 palmitoylation and autophagy/MAPK readouts","pmids":["41603995","41566717"],"confidence":"Medium","gaps":["No crystal/cryo-EM structure confirming His72 as an activation site","ZEB-2 deubiquitinase linking palmitoylation to stability not identified"]},{"year":null,"claim":"How ZDHHC4 selects among its many substrates and whether outcomes (membrane targeting, nuclear import, stabilization, or lysosomal degradation) are dictated by adaptor recruitment, localization, or substrate context remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of ZDHHC4 substrate engagement","Catalytic and regulatory residues largely uncharacterized beyond His72","Redundancy with ZDHHC5/ZDHHC8 not systematically dissected"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,7,6,2,3,4,9]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,7,9]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,7,6,2,3,9]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[6,8,11]}],"complexes":[],"partners":["CPT1A","MAVS","TRPV1","CD36","KAI1","DRD2","GSK3B","ZEB2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NPG8","full_name":"Palmitoyltransferase ZDHHC4","aliases":["Zinc finger DHHC domain-containing protein 4","DHHC-4","Zinc finger protein 374"],"length_aa":344,"mass_kda":39.8,"function":"Palmitoyltransferase that catalyzes the addition of palmitate onto protein substrates including the D(2) dopamine receptor DRD2, GSK3B or MAVS. Mediates GSK3B palmitoylation to prevent its AKT1-mediated phosphorylation leading to activation of the STAT3 signaling pathway (PubMed:35606353). Also catalyzes MAVS palmitoylation which promotes its stabilization and activation by inhibiting 'Lys-48'- but facilitating 'Lys-63'-linked ubiquitination (PubMed:38016475)","subcellular_location":"Endoplasmic reticulum membrane; Golgi apparatus membrane; Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q9NPG8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ZDHHC4","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ZDHHC4","total_profiled":1310},"omim":[{"mim_id":"621547","title":"ZDHHC PALMITOYLTRANSFERASE 4; ZDHHC4","url":"https://www.omim.org/entry/621547"},{"mim_id":"617150","title":"ZDHHC PALMITOYLTRANSFERASE 3; ZDHHC3","url":"https://www.omim.org/entry/617150"},{"mim_id":"608784","title":"ZDHHC PALMITOYLTRANSFERASE 8; ZDHHC8","url":"https://www.omim.org/entry/608784"},{"mim_id":"605599","title":"LYSOPHOSPHOLIPASE I; LYPLA1","url":"https://www.omim.org/entry/605599"},{"mim_id":"605004","title":"GLYCOGEN SYNTHASE KINASE 3-BETA; GSK3B","url":"https://www.omim.org/entry/605004"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ZDHHC4"},"hgnc":{"alias_symbol":["FLJ10479","ZNF374"],"prev_symbol":[]},"alphafold":{"accession":"Q9NPG8","domains":[{"cath_id":"-","chopping":"72-122_191-296_319-332","consensus_level":"medium","plddt":89.0337,"start":72,"end":332},{"cath_id":"-","chopping":"125-173","consensus_level":"medium","plddt":94.4065,"start":125,"end":173}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPG8","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPG8-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NPG8-F1-predicted_aligned_error_v6.png","plddt_mean":84.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ZDHHC4","jax_strain_url":"https://www.jax.org/strain/search?query=ZDHHC4"},"sequence":{"accession":"Q9NPG8","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NPG8.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NPG8/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NPG8"}},"corpus_meta":[{"pmid":"38016475","id":"PMC_38016475","title":"CPT1A induction following epigenetic perturbation promotes MAVS palmitoylation and activation to potentiate antitumor immunity.","date":"2023","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/38016475","citation_count":53,"is_preprint":false},{"pmid":"34528721","id":"PMC_34528721","title":"Palmitoylation of SARS-CoV-2 S protein is critical for S-mediated syncytia formation and virus entry.","date":"2021","source":"Journal of medical virology","url":"https://pubmed.ncbi.nlm.nih.gov/34528721","citation_count":50,"is_preprint":false},{"pmid":"34530889","id":"PMC_34530889","title":"KAI1(CD82) is a key molecule to control angiogenesis and switch angiogenic milieu to quiescent state.","date":"2021","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34530889","citation_count":43,"is_preprint":false},{"pmid":"26535572","id":"PMC_26535572","title":"Effect of C-Terminal S-Palmitoylation on D2 Dopamine Receptor Trafficking and Stability.","date":"2015","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26535572","citation_count":42,"is_preprint":false},{"pmid":"39047111","id":"PMC_39047111","title":"Atf3-mediated metabolic reprogramming in hepatic macrophage orchestrates metabolic dysfunction-associated steatohepatitis.","date":"2024","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/39047111","citation_count":32,"is_preprint":false},{"pmid":"35606353","id":"PMC_35606353","title":"GSK3β palmitoylation mediated by ZDHHC4 promotes tumorigenicity of glioblastoma stem cells in temozolomide-resistant glioblastoma through the EZH2-STAT3 axis.","date":"2022","source":"Oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/35606353","citation_count":28,"is_preprint":false},{"pmid":"27956064","id":"PMC_27956064","title":"Novel genes in brain tissues of EAE-induced normal and obese mice: Upregulation of metal ion-binding protein genes in obese-EAE mice.","date":"2016","source":"Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/27956064","citation_count":20,"is_preprint":false},{"pmid":"35150145","id":"PMC_35150145","title":"Ppt1-deficiency dysregulates lysosomal Ca++ homeostasis contributing to pathogenesis in a mouse model of CLN1 disease.","date":"2022","source":"Journal of inherited metabolic disease","url":"https://pubmed.ncbi.nlm.nih.gov/35150145","citation_count":17,"is_preprint":false},{"pmid":"40357580","id":"PMC_40357580","title":"FoxO1-zDHHC4-CD36 S-Acylation Axis Drives Metabolic Dysfunction in Diabetes.","date":"2025","source":"Circulation research","url":"https://pubmed.ncbi.nlm.nih.gov/40357580","citation_count":11,"is_preprint":false},{"pmid":"39563863","id":"PMC_39563863","title":"Research progress on S-palmitoylation modification mediated by the ZDHHC family in glioblastoma.","date":"2024","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/39563863","citation_count":9,"is_preprint":false},{"pmid":"39528731","id":"PMC_39528731","title":"Palmitoylation by ZDHHC4 inhibits TRPV1-mediated nociception.","date":"2024","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/39528731","citation_count":4,"is_preprint":false},{"pmid":"41306774","id":"PMC_41306774","title":"EVA1A Regulates Hepatic Lipid Homeostasis by Modulating CD36 Expression and Its Palmitoylation.","date":"2025","source":"Research (Washington, D.C.)","url":"https://pubmed.ncbi.nlm.nih.gov/41306774","citation_count":2,"is_preprint":false},{"pmid":"40753321","id":"PMC_40753321","title":"ZDHHC4 Influences Obsessive-Compulsive Disorder Risk Through Imaging-Derived Phenotypes: A Mendelian Randomization Study.","date":"2025","source":"Journal of molecular neuroscience : MN","url":"https://pubmed.ncbi.nlm.nih.gov/40753321","citation_count":0,"is_preprint":false},{"pmid":"41603995","id":"PMC_41603995","title":"ZDHHC4-mediated ZEB-2 S-palmitoylation promotes melanoma epithelial-mesenchymal transition via deubiquitinating and stabilizing ZEB-2.","date":"2026","source":"Cellular and molecular life sciences : CMLS","url":"https://pubmed.ncbi.nlm.nih.gov/41603995","citation_count":0,"is_preprint":false},{"pmid":"41566717","id":"PMC_41566717","title":"Lactucopicrin promotes the autophagic degradation of MAP2K4/MKK4 by mediating CCDC50 palmitoylation to alleviate osteoarthritis progression.","date":"2026","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/41566717","citation_count":0,"is_preprint":false},{"pmid":"41049004","id":"PMC_41049004","title":"Inflammatory Bowel Disease Mediates the Causal Relationship Between Gut Microbiota and Colorectal Cancer: Identification of Therapeutic Targets and Predictive Modeling.","date":"2025","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/41049004","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10943,"output_tokens":3218,"usd":0.04055,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10611,"output_tokens":3262,"usd":0.067303,"stage2_stop_reason":"end_turn"},"total_usd":0.107853,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2023,\n      \"finding\": \"CPT1A recruits the ER-localized ZDHHC4 to catalyze MAVS palmitoylation at Cys79, which promotes MAVS stabilization and activation by inhibiting K48-linked ubiquitination while facilitating K63-linked ubiquitination, thereby sustaining IFN-I response.\",\n      \"method\": \"Mechanistic cell-based assays, co-immunoprecipitation, palmitoylation assays, ubiquitination analysis, site-specific mutagenesis\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, site-specific mutagenesis (Cys79), multiple orthogonal readouts (palmitoylation, K48/K63 ubiquitination, IFN-I response), functionally validated in vivo\",\n      \"pmids\": [\"38016475\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZDHHC4 physically associates with SARS-CoV-2 S protein and its overexpression promotes palmitoylation of the S protein cytoplasmic tail, which is required for S-mediated syncytia formation and pseudovirus entry but not plasma membrane targeting.\",\n      \"method\": \"Co-immunoprecipitation, palmitoylation assay, overexpression screen with multiple ZDHHC family members, pseudovirus entry assay, syncytia formation assay\",\n      \"journal\": \"Journal of medical virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with functional readouts, single lab, multiple orthogonal assays but no site-specific mutagenesis for ZDHHC4 specifically\",\n      \"pmids\": [\"34528721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZDHHC4 palmitoylates KAI1 (CD82), which is required for KAI1 localization to the membrane surface; membrane-localized KAI1 then induces LIF via the Src/p53 pathway to suppress angiogenesis.\",\n      \"method\": \"KAI1 knockout mice, in vitro palmitoylation assay, in vivo angiogenesis models, mechanistic pathway analysis\",\n      \"journal\": \"Journal of hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional KO model with defined cellular phenotype, palmitoylation linked to localization and pathway activation, single lab\",\n      \"pmids\": [\"34530889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"ZDHHC4 physically interacts with the D2 dopamine receptor (D2R) and affects its palmitoylation status; palmitoylation at C443 is required for D2R plasma membrane expression and protein stability.\",\n      \"method\": \"Membrane yeast two-hybrid (MYTH) screen, co-immunoprecipitation, bioorthogonal click chemistry palmitoylation assay, site-directed mutagenesis\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MYTH screen confirmed by Co-IP, palmitoylation assay with mutagenesis, single lab\",\n      \"pmids\": [\"26535572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZDHHC4 palmitoylates GSK3β at Cys14, which increases GSK3β activity (decreasing p-Ser9, increasing p-Tyr216) and activates the EZH2-STAT3 signaling axis to promote GBM stem cell self-renewal and temozolomide resistance.\",\n      \"method\": \"Palmitoylation assay, site-directed mutagenesis (Cys14), knockdown/overexpression, phosphorylation analysis, STAT3/EZH2 pathway readouts in GBM stem cells\",\n      \"journal\": \"Oncogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-specific mutagenesis combined with functional pathway readouts, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"35606353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZDHHC4 (together with ZDHHC8) catalyzes S-palmitoylation of NFATC4, which is required for NFATC4 trafficking from the cytoplasm to the nucleus; in Ppt1-/- mice reduced ZDHHC4/ZDHHC8 levels lower nuclear palmitoylated NFATC4, inhibiting IP3R1 expression and dysregulating lysosomal Ca2+ homeostasis.\",\n      \"method\": \"Identification of ZDHHC4 and ZDHHC8 as NFATC4 palmitoyl acyltransferases, subcellular fractionation, palmitoylation assay in Ppt1-/- mouse model, IP3R1 overexpression rescue experiment\",\n      \"journal\": \"Journal of inherited metabolic disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — palmitoylation assay linked to subcellular trafficking and downstream functional consequence, genetic model with rescue, single lab\",\n      \"pmids\": [\"35150145\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FoxO1 transcriptionally upregulates ZDHHC4 expression in the diabetic heart; ZDHHC4 S-acylates CD36, promoting its sarcolemmal localization; genetic silencing of ZDHHC4 decreases CD36 S-acylation and sarcolemmal content, reducing fatty acid oxidation and triglyceride storage and improving cardiac function.\",\n      \"method\": \"ChIP-seq, ChIP-qPCR, luciferase reporter assay, siRNA/shRNA silencing, cardiomyocyte-specific FoxO1 knockout mice, pharmacological FoxO1 inhibition, S-acylation assay across multiple species (rat, db/db mouse, pig, human iPSC-CMs)\",\n      \"journal\": \"Circulation research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (ChIP-seq, luciferase, genetic KO, pharmacological inhibition), cross-species validation, direct mechanistic chain from FoxO1 to ZDHHC4 to CD36 palmitoylation to functional metabolic phenotype\",\n      \"pmids\": [\"40357580\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ZDHHC4 physically interacts with TRPV1 and catalyzes S-palmitoylation at C157, C362, C390, and C715, promoting TRPV1 degradation via the lysosome pathway; this modification is counterbalanced by depalmitoylase APT1, and the net palmitoylation state regulates inflammatory pain relief.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis of TRPV1 cysteines, palmitoylation assay, lysosomal degradation assay, electrophysiology, in vivo pain behavior\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — site-specific mutagenesis of four palmitoylation sites, electrophysiology, in vivo evidence, and identification of the opposing depalmitoylase, multiple orthogonal methods in one study\",\n      \"pmids\": [\"39528731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ATF3 in macrophages increases ZDHHC4/5-mediated CD36 palmitoylation at C3, C7, C464, and C466, promoting CD36 plasma membrane localization; this contributes to regulation of fatty acid oxidation and protection against MASH.\",\n      \"method\": \"Overexpression/ablation of Atf3 in macrophages, palmitoylation assay with site identification, subcellular fractionation, in vivo MASH mouse model\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — site-specific palmitoylation with functional localization readout, but ZDHHC4 and ZDHHC5 not individually resolved; single lab\",\n      \"pmids\": [\"39047111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"ZDHHC4 palmitoylates ZEB-2 at C478 (identified by mass spectrometry); ZEB-2 interacts with ZDHHC4 through its N-terminal sequences; palmitoylation promotes ZEB-2 deubiquitination and protein stability, which is required for EMT in melanoma cells.\",\n      \"method\": \"Mass spectrometry substrate identification, Co-immunoprecipitation, site-directed mutagenesis (C478), knockdown functional assays (proliferation, migration, invasion), ubiquitination assay\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS substrate ID confirmed by Co-IP and mutagenesis, functional knockdown with defined phenotype, single lab\",\n      \"pmids\": [\"41603995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"LCP (lactucopicrin) directly binds to His72 of ZDHHC4, boosting its enzymatic activity; activated ZDHHC4 catalyzes palmitoylation of CCDC50, facilitating selective autophagic degradation of MAP2K4/MKK4, suppressing MAPK/JNK signaling and chondrocyte senescence in osteoarthritis.\",\n      \"method\": \"DARTS (drug affinity responsive target stability) structural binding assay, palmitoylation assay, autophagy flux assay, MAPK/JNK signaling readouts, in vivo OA mouse model\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding to ZDHHC4 His72 identified, palmitoylation linked to autophagy and signaling cascade, in vivo validation, single lab\",\n      \"pmids\": [\"41566717\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVA1A deficiency transcriptionally enhances ZDHHC4/5 expression while repressing the depalmitoylase APT1, resulting in increased CD36 palmitoylation and plasma membrane localization; this promotes fatty acid uptake and impairs β-oxidation in hepatocytes.\",\n      \"method\": \"Hepatocyte-specific knockout of Eva1a, overexpression rescue, palmitoylation assay, subcellular fractionation, transcriptional analysis of ZDHHC4/5 and APT1\",\n      \"journal\": \"Research (Washington, D.C.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with functional metabolic phenotype and palmitoylation readout, but ZDHHC4 and ZDHHC5 not individually resolved; single lab\",\n      \"pmids\": [\"41306774\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ZDHHC4 is an ER/membrane-localized palmitoyl acyltransferase that catalyzes S-palmitoylation of diverse substrates—including MAVS (Cys79), TRPV1 (C157/C362/C390/C715), GSK3β (Cys14), ZEB-2 (C478), NFATC4, CD36, KAI1, D2R, and CCDC50—to regulate their stability, subcellular trafficking, and signaling activity in contexts ranging from innate immune activation and pain modulation to metabolic regulation, cancer stemness, and EMT, with its own expression controlled transcriptionally by FoxO1 in metabolic disease.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ZDHHC4 is an ER/membrane-localized palmitoyl acyltransferase that catalyzes S-palmitoylation of a structurally diverse set of substrates to control their stability, subcellular trafficking, and downstream signaling across innate immunity, pain, metabolism, and cancer [#0, #7, #6]. A recurrent mechanistic theme is that ZDHHC4-mediated palmitoylation governs substrate fate through two routes: directing membrane or nuclear localization, and modulating ubiquitin-dependent stability. In innate immunity, ZDHHC4 is recruited by CPT1A to palmitoylate MAVS at Cys79, blocking K48-linked ubiquitination while favoring K63-linked ubiquitination to stabilize MAVS and sustain the type I interferon response [#0]; an analogous stabilization mechanism operates on ZEB-2 (palmitoylation at C478 promoting deubiquitination) to drive EMT in melanoma [#9]. ZDHHC4 also controls trafficking-dependent activity: it palmitoylates KAI1/CD82 and the D2 dopamine receptor to direct their plasma-membrane localization and stability [#2, #3], drives nuclear translocation of NFATC4 [#5], and—in metabolic tissues—S-acylates CD36 to promote its sarcolemmal/plasma-membrane localization, fatty-acid uptake and oxidation, with ZDHHC4 expression itself transcriptionally upregulated by FoxO1 in the diabetic heart [#6, #8, #11]. Conversely, ZDHHC4 palmitoylation of TRPV1 at C157/C362/C390/C715 targets the channel for lysosomal degradation, a modification opposed by the depalmitoylase APT1 and tuning inflammatory pain [#7]. Through palmitoylation of GSK3\\u03b2 at Cys14 it activates the EZH2-STAT3 axis in glioblastoma stem cells [#4], and palmitoylation of CCDC50 links it to selective autophagy of MAP2K4 and suppression of MAPK/JNK signaling [#10]. Its enzymatic activity is regulated at the residue level, being boosted by the small molecule lactucopicrin binding His72 [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2015,\n      \"claim\": \"Established ZDHHC4 as a bona fide acyltransferase acting on a membrane receptor, addressing whether it physically engages and modifies substrates to control their surface expression.\",\n      \"evidence\": \"Membrane yeast two-hybrid screen, Co-IP and click-chemistry palmitoylation assay with D2R cysteine mutagenesis\",\n      \"pmids\": [\"26535572\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab without reciprocal in vitro reconstitution of ZDHHC4 enzymatic activity\", \"Did not define ZDHHC4 catalytic residues\", \"Trafficking mechanism downstream of palmitoylation not resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended ZDHHC4 substrate repertoire to a tetraspanin and a viral glycoprotein, showing palmitoylation controls membrane localization and downstream signaling versus viral fusion.\",\n      \"evidence\": \"KAI1 knockout mice with palmitoylation and angiogenesis assays; Co-IP and pseudovirus/syncytia assays for SARS-CoV-2 S protein\",\n      \"pmids\": [\"34530889\", \"34528721\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No ZDHHC4-specific site mutagenesis for the S protein\", \"Direct catalytic role not separated from family redundancy in the viral study\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Showed ZDHHC4 palmitoylation can both activate a kinase and license a transcription factor's nuclear entry, broadening its role to oncogenic and Ca2+-homeostasis signaling.\",\n      \"evidence\": \"Site-directed mutagenesis (GSK3\\u03b2 Cys14) with EZH2-STAT3 readouts in GBM stem cells; NFATC4 palmitoylation and fractionation in Ppt1-/- mice with IP3R1 rescue\",\n      \"pmids\": [\"35606353\", \"35150145\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"NFATC4 modification shared with ZDHHC8 and not individually resolved\", \"Direct enzyme-substrate kinetics not established\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined the highest-confidence mechanism: CPT1A-recruited ZDHHC4 palmitoylates MAVS at Cys79 to redirect ubiquitin signals (suppressing K48, favoring K63) and sustain interferon responses.\",\n      \"evidence\": \"Reciprocal Co-IP, Cys79 mutagenesis, palmitoylation and K48/K63 ubiquitination assays, in vivo IFN-I readouts\",\n      \"pmids\": [\"38016475\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of CPT1A-ZDHHC4 recruitment not solved\", \"Whether MAVS palmitoylation is reversed by a specific depalmitoylase unaddressed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected ZDHHC4 to substrate degradation rather than stabilization, and to physiological metabolism, establishing context-dependent outcomes of palmitoylation.\",\n      \"evidence\": \"TRPV1 four-site mutagenesis with lysosomal degradation, electrophysiology and pain behavior plus APT1 opposition; ATF3-driven CD36 palmitoylation with fractionation in a MASH model\",\n      \"pmids\": [\"39528731\", \"39047111\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"CD36 study did not individually resolve ZDHHC4 from ZDHHC5\", \"How substrate identity dictates lysosomal versus stabilizing fate is unexplained\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Placed ZDHHC4 within transcriptional regulatory circuits controlling cardiac and hepatic lipid handling via CD36 palmitoylation.\",\n      \"evidence\": \"FoxO1 ChIP-seq/luciferase and cardiomyocyte FoxO1 KO with cross-species S-acylation assays; hepatocyte Eva1a KO with ZDHHC4/5 and APT1 transcriptional analysis\",\n      \"pmids\": [\"40357580\", \"41306774\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"EVA1A study did not resolve ZDHHC4 from ZDHHC5\", \"Direct ZDHHC4 enzymatic specificity for CD36 cysteines not isolated from family members\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified a stabilizing substrate (ZEB-2) and a pharmacological activation handle (His72), advancing ZDHHC4 as a druggable enzyme with defined regulatory residues.\",\n      \"evidence\": \"Mass spectrometry, Co-IP and C478 mutagenesis for ZEB-2; DARTS binding to His72 by lactucopicrin with CCDC50 palmitoylation and autophagy/MAPK readouts\",\n      \"pmids\": [\"41603995\", \"41566717\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No crystal/cryo-EM structure confirming His72 as an activation site\", \"ZEB-2 deubiquitinase linking palmitoylation to stability not identified\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How ZDHHC4 selects among its many substrates and whether outcomes (membrane targeting, nuclear import, stabilization, or lysosomal degradation) are dictated by adaptor recruitment, localization, or substrate context remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of ZDHHC4 substrate engagement\", \"Catalytic and regulatory residues largely uncharacterized beyond His72\", \"Redundancy with ZDHHC5/ZDHHC8 not systematically dissected\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 7, 6, 2, 3, 4, 9]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 7, 9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 7, 6, 2, 3, 9]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [6, 8, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"CPT1A\", \"MAVS\", \"TRPV1\", \"CD36\", \"KAI1\", \"DRD2\", \"GSK3B\", \"ZEB2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}