{"gene":"PI4KB","run_date":"2026-06-10T06:43:35","timeline":{"discoveries":[{"year":2011,"finding":"ACBD3 (a Golgi-resident protein) interacts with PI4KB and recruits it to Aichi virus RNA replication sites, forming a viral protein/ACBD3/PI4KB complex that synthesizes PI4P at replication sites essential for viral RNA replication.","method":"Co-immunoprecipitation, knockdown of ACBD3 or PI4KB (loss-of-function with viral replication readout), colocalization imaging","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction mapping, knockdown phenotype, colocalization; independently replicated across multiple labs in subsequent studies","pmids":["22124328"],"is_preprint":false},{"year":2016,"finding":"NMR structure of the PI4KB–ACBD3 complex was determined; ACBD3 recruits PI4KB to membranes both in vitro and in vivo and membrane recruitment increases PI4KB enzymatic (lipid kinase) activity; the ACBD3:PI4KB complex is essential for proper Golgi function.","method":"NMR structure determination, in vitro membrane recruitment assay, in vivo localization, enzymatic activity assay","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — NMR structure plus in vitro reconstitution with enzymatic activity measurement plus in vivo validation in a single study","pmids":["27009356"],"is_preprint":false},{"year":2016,"finding":"Crystal structure of the ACBD3 GOLD domain revealed a unique N-terminus mediating interaction with Aichi virus 3A protein; 3A directly activates PI4KIIIβ lipid kinase activity, and this activation is sensitized by ACBD3; complex-disrupting mutations in ACBD3 and PI4KIIIβ abrogated this sensitization. Interfaces were mapped by hydrogen-deuterium exchange mass spectrometry (HDX-MS).","method":"Biochemical reconstitution, crystal structure determination, HDX-MS interface mapping, mutagenesis, lipid kinase activity assay","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure, in vitro reconstitution, mutagenesis, and HDX-MS in a single rigorous study","pmids":["27989622"],"is_preprint":false},{"year":2016,"finding":"PI4KB inhibitors bind in the ATP-binding site of PI4KB; crystallographic analysis confirmed the binding mode and explains their inhibitory activity against PI4KB, with selectivity over other kinases.","method":"X-ray crystallography, enzyme inhibition assays, antiviral assays","journal":"Journal of medicinal chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structures plus in vitro enzymatic assays, single study but multiple orthogonal methods","pmids":["28004945"],"is_preprint":false},{"year":2017,"finding":"PI4KB forms a tight 2:2 complex with 14-3-3 proteins upon phosphorylation; 14-3-3 binding does not directly modulate PI4KB enzymatic activity but protects PI4KB from proteolytic degradation in vitro; structural analysis showed 14-3-3 binding does not interfere with ACBD3-mediated membrane recruitment.","method":"Biophysical characterization (SAXS, fluorescence methods), proteolysis protection assay, enzymatic activity assay, structural analysis","journal":"Journal of structural biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple orthogonal biophysical methods plus biochemical validation of structural predictions in a single study","pmids":["28864297"],"is_preprint":false},{"year":2019,"finding":"ACBD3 is an essential mediator of enterovirus 3A-dependent PI4KB recruitment to replication organelles; ACBD3 knockout prevented PI4KB recruitment and impaired 3A Golgi localization; a PI4KB mutant unable to bind ACBD3 failed to restore replication in PI4KB-KO cells; the ACB and CAR domains of ACBD3 are dispensable for PI4KB recruitment.","method":"ACBD3 and PI4KB knockout cells, reconstitution with wild-type and mutant proteins, fluorescence microscopy localization, virus replication assays","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple KO cell lines reconstituted with defined mutants, replicated across multiple enterovirus species, multiple orthogonal readouts","pmids":["30755512"],"is_preprint":false},{"year":2019,"finding":"c10orf76 forms a complex with PI4KB mediated by the kinase linker of PI4KB; PKA-dependent phosphorylation modulates complex formation; PI4KB is required for membrane recruitment of c10orf76 to the Golgi; the intact c10orf76–PI4KB complex increases Golgi PI4P levels and is required for replication of specific enteroviruses; c10orf76 contributes to Arf1 activation at the Golgi.","method":"HDX-MS complex characterization, complex-disrupting mutagenesis, PI4P level measurement, virus replication assay, localization studies","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — HDX-MS plus mutagenesis plus functional PI4P and viral replication readouts in a single rigorous study","pmids":["31829496"],"is_preprint":false},{"year":2019,"finding":"PI4KB forms highly flexible heterocomplexes including ACBD3, 14-3-3, and Rab11 proteins; the 14-3-3:PI4KB:Rab11 complex has 2:1:1 stoichiometry; the ACBD3:PI4KB complex exists in very compact and extended conformations; membrane is necessary for formation of the ACBD3:PI4KB:Rab11 complex at physiological concentrations.","method":"Small angle X-ray scattering (SAXS), in vitro reconstitution","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — SAXS structural characterization and in vitro reconstitution, single lab, single study","pmids":["30679637"],"is_preprint":false},{"year":2006,"finding":"PI4K92 (PI4KB) phosphorylated at Ser-294 localizes exclusively to the Golgi and this phosphorylation increases lipid kinase activity; PI4K92 phosphorylated at Ser-496 and Thr-504 localizes to nuclear speckles dynamically and is required for cell viability (microinjection of anti-pSer-496 antibody caused apoptosis).","method":"Anti-phosphopeptide antibodies, indirect immunofluorescence, microinjection, lipid kinase activity assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct subcellular localization with site-specific antibodies plus functional readout (kinase activity, microinjection phenotype), single lab","pmids":["16606619"],"is_preprint":false},{"year":2001,"finding":"Recombinant PI4K92 (PI4KB) produces PtdIns4P as product; is characterized as a type III PI4K (high Km for ATP and PtdIns in millimolar range, IC50 ~300 nM for Wortmannin); multiple phosphorylation sites mapped by MALDI-MS and LC-MS/MS at S258, T263, S266, S277, S294, T423, S496, T504 within a designated phosphorylation domain.","method":"Recombinant protein expression, in vitro lipid kinase assay, MALDI-MS, LC-MS/MS phosphorylation site mapping","journal":"European journal of biochemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro enzymatic characterization plus mass spectrometry phosphorylation site mapping, directly on recombinant PI4KB","pmids":["11277933"],"is_preprint":false},{"year":2022,"finding":"ARL5A and ARL5B GTPases interact with PI4KB at the trans-Golgi and recruit it, thereby promoting PI4KB's function in PI4P synthesis and protein secretion.","method":"Proximity biotinylation (miniTurboID), quantitative mass spectrometry, protein interaction assays","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proximity labeling plus functional PI4P and secretion readouts, but PI4KB is one of many interactors identified in a broader Arf-family screen","pmids":["35844135"],"is_preprint":false},{"year":2024,"finding":"ARMH3 (C10orf76) is an effector of active ARL5 at the TGN; ARMH3 activates PI4KB to account for the main pool of PI4P at the TGN; this contributes to GOLPH3 recruitment and glycan modifications; the SYS1-ARFRP1-ARL5-ARMH3 axis regulates PI4KB-dependent PI4P generation at the TGN.","method":"Proximity biotinylation, protein interaction assays (active vs. inactive ARL5 mutants), PI4P level measurement, knockdown/knockout with functional readouts (GOLPH3 localization, glycan modification)","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (proximity labeling, direct binding assays with GTPase mutants, functional PI4P and downstream readouts), rigorous study","pmids":["39580461"],"is_preprint":false},{"year":2023,"finding":"ARMH3 interacts with STING at the Golgi upon cGAMP stimulation and recruits PI4KB to synthesize PI4P, which directs STING Golgi-to-endosome trafficking via PI4P-binding proteins AP-1 and GGA2; disrupting PI4P-dependent transport impaired STING activation; elevated cellular PI4P was sufficient for cGAS-independent STING activation.","method":"Genome-wide CRISPR-Cas9 screen, co-immunoprecipitation, PI4P measurement, RNAi of PI4P-binding proteins, in vivo mouse model (Armh3fl/fl LyzCre/Cre)","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide unbiased screen plus mechanistic validation with co-IP, multiple RNAi knockdowns, PI4P manipulation, and in vivo mouse model","pmids":["36921576"],"is_preprint":false},{"year":2023,"finding":"The C10orf76–PI4KB axis generates a specific pool of PI4P at distal Golgi subregions; CERT preferentially utilizes PI4P generated by PI4KB recruited via C10orf76 (not ACBD3) for ER-to-Golgi ceramide trafficking; C10orf76 localizes predominantly at distal Golgi where sphingomyelin synthesis occurs, while ACBD3 is at more proximal regions.","method":"Genome-wide screening, PI4P measurement, super-resolution microscopy, ceramide transport assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide unbiased screen plus super-resolution microscopy plus lipid transfer functional assay, multiple orthogonal methods","pmids":["37195633"],"is_preprint":false},{"year":2018,"finding":"RAB30 interacts with PI4KB and recruits it to the Golgi apparatus and GAS-containing autophagosome-like vacuoles (GcAVs); PI4KB knockout suppressed autophagy by inhibiting GcAV formation, resulting in increased GAS survival; PtdIns4P is crucial for GcAV formation.","method":"Co-immunoprecipitation (RAB30–PI4KB interaction), PI4KB knockout cells, autophagy/GcAV formation assay, PI4P depletion/repletion experiments","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP interaction plus KO phenotype plus PI4P functional rescue, single lab","pmids":["30290718"],"is_preprint":false},{"year":2019,"finding":"PI4KB is recruited to inclusion bodies formed by ER membrane remodeling during HPIV3 infection; the HPIV3 phosphoprotein (P) recruits PI4KB to IBs to generate PI4P, creating a PI4P-enriched microenvironment that promotes viral replication; HRSV nucleoprotein similarly recruits PI4KB to IBs.","method":"Co-immunoprecipitation, PI4P measurement, colocalization imaging, knockdown/loss-of-function viral replication assays","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, PI4P measurement, and functional replication readouts, single lab","pmids":["31747597"],"is_preprint":false},{"year":2017,"finding":"Under PI4KB inhibition, PI4KB activity has distinct functions in both proteolytic processing of the enteroviral polyprotein and in replication organelle (RO) biogenesis; a PI4KB-inhibitor-resistant escape mutation corrects a proteolytic processing defect, revealing an unexpected role of PI4KB in viral polyprotein processing.","method":"PI4KB inhibitor-resistant mutant virus selection, electron tomography of ROs, viral polyprotein processing analysis","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — resistance mutation mapping plus structural (EM) and biochemical readouts, single lab","pmids":["29045829"],"is_preprint":false},{"year":2020,"finding":"SC-specific genetic inactivation of PI4KB in mice disrupts Golgi morphology in Schwann cells, causes disappearance of OSBP at cis-Golgi and loss of GOLPH3 from the entire Golgi, reduces cholesterol and sphingomyelin in sciatic nerves, reduces caveolae, and results in thinner myelin, abnormal nodes of Ranvier, and impaired engulfment of nerve fibers; demonstrating PI4KB Golgi function is required for myelination through lipid metabolism and protein glycosylation control.","method":"Conditional PI4KB knockout mice (SC-specific), electron microscopy, immunofluorescence, lipid analysis, nerve conduction velocity measurement","journal":"PNAS","confidence":"High","confidence_rationale":"Tier 2 / Strong — conditional KO mouse model with multiple orthogonal cellular, biochemical, and physiological readouts","pmids":["33106410"],"is_preprint":false},{"year":2025,"finding":"Oncogenic KRAS induces a non-canonical autophagy pathway (RINCAA) involving a P38-ULK1-PI4KB-WIPI2 signaling cascade; ULK1 phosphorylates PI4KB at S256 and T263, triggering PI4P production, ATG8ylation, and non-canonical autophagosome formation; inhibiting these phosphorylation sites reduces RINCAA activity and tumor growth in xenograft and KPC pancreatic cancer models.","method":"Genetic knockouts, phosphorylation site mutagenesis, PI4P measurement, autophagy flux assays, in vivo xenograft and KPC mouse models","journal":"Cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — mutagenesis of specific phosphorylation sites plus multiple in vivo cancer models plus multiple orthogonal autophagy readouts","pmids":["40055523"],"is_preprint":false},{"year":2021,"finding":"ANXA2 interacts with both EV71 3D polymerase and PI4KB; the annexin domain of ANXA2 mediates 3D binding; ANXA2 localizes to replication organelles; ANXA2 overexpression stimulates PI4P production and promotes the PI4KB–3D interaction, facilitating formation of a higher-order ANXA2–PI4KB–3D protein complex at the viral replication site.","method":"Co-immunoprecipitation, ANXA2 knockout, PI4P level measurement, colocalization imaging","journal":"Virologica Sinica","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, KO with PI4P measurement, colocalization, single lab","pmids":["34196914"],"is_preprint":false},{"year":2025,"finding":"SCAMP5 is a binding partner of PI4KB/PI4KIIIβ at the TGN; SCAMP5 controls PI4KB recruitment to the TGN and subsequent PI4P production; PI4P is essential for AP-4 recruitment; SCAMP5 depletion disrupts AP-4-mediated ATG9A trafficking to presynaptic sites, impairing presynaptic autophagy.","method":"Co-immunoprecipitation (SCAMP5–PI4KB interaction), SCAMP5 knockdown with PI4P measurement, AP-4 localization, ATG9A trafficking assay, autophagosome formation assay","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus functional PI4P and trafficking readouts, single lab","pmids":["40958389"],"is_preprint":false},{"year":2014,"finding":"HCV NS5A competes with PI4KB for binding to ACBD3 in a genotype-dependent manner; ACBD3 binds GT1b NS5A with higher affinity than GT2a NS5A, correlating with higher PI4KB/PI4P colocalization in GT1b-infected cells; NS5A displaces PI4KB from ACBD3 to allow PI4KB relocation to replication sites.","method":"Co-immunoprecipitation (competitive binding), colocalization imaging","journal":"Antiviral research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP competition assay plus colocalization, single lab, two orthogonal approaches","pmids":["24792752"],"is_preprint":false},{"year":2016,"finding":"A yeast 14-3-3 protein crystal structure bound to a phosphopeptide derived from human PI4KB was determined, demonstrating that the 14-3-3 recognition mode for PI4KB phosphopeptide is highly evolutionarily conserved.","method":"X-ray crystallography of 14-3-3 bound to PI4KB-derived phosphopeptide","journal":"Acta crystallographica Section F","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — crystal structure with PI4KB peptide, single study, structural only without functional mutagenesis of PI4KB itself","pmids":["27827352"],"is_preprint":false}],"current_model":"PI4KB (PI4K IIIβ) is a Golgi/TGN-localized lipid kinase that phosphorylates phosphatidylinositol to produce PI4P; it is recruited to the Golgi by multiple adaptors (ACBD3, C10orf76/ARMH3, RAB30, ARL5A/B, SCAMP5), regulated by 14-3-3 protein binding (which stabilizes but does not directly alter its activity) and by phosphorylation at multiple sites (including PKD-mediated and ULK1-mediated sites), and generates distinct PI4P pools that control membrane trafficking, ceramide transport, Golgi function, myelination, STING antiviral signaling, and non-canonical autophagy; PI4KB is also hijacked by numerous RNA viruses through viral proteins that co-opt ACBD3 or other adaptors to redirect PI4KB to viral replication organelles."},"narrative":{"mechanistic_narrative":"PI4KB (PI4KIIIβ) is a type III phosphatidylinositol 4-kinase that phosphorylates phosphatidylinositol to generate PI4P, and the Golgi/trans-Golgi PI4P pools it produces control membrane trafficking, secretion, lipid transport, antiviral signaling, autophagy, and myelination [PMID:11277933, PMID:39580461, PMID:33106410]. Catalytically, PI4KB is a high-Km type III kinase whose activity is tuned both by membrane recruitment and by phosphorylation at a defined cluster of sites (S294 increasing kinase activity, with additional sites including S496/T504 governing localization) [PMID:16606619, PMID:11277933]. Its function depends on recruitment to specific membranes by a panel of adaptors: ACBD3 binds PI4KB and, upon membrane recruitment, stimulates its lipid kinase activity to support Golgi function [PMID:27009356, PMID:27989622], while ARL5A/B GTPases and their effector ARMH3/C10orf76 define a SYS1–ARFRP1–ARL5–ARMH3 axis that generates the main TGN PI4P pool driving GOLPH3 recruitment, glycosylation, and secretion [PMID:31829496, PMID:35844135, PMID:39580461]; RAB30 and SCAMP5 provide additional recruitment routes [PMID:30290718, PMID:40958389]. PI4KB partitions into distinct subcellular PI4P pools, with the C10orf76/ARMH3-recruited pool at distal Golgi feeding CERT-mediated ceramide transport, separate from the ACBD3-associated proximal pool [PMID:37195633]. PI4KB is regulated post-translationally by phosphorylation-dependent 14-3-3 binding, which forms a 2:2 complex that protects PI4KB from proteolysis without altering its catalytic activity or blocking ACBD3-mediated recruitment [PMID:28864297, PMID:30679637]. Beyond constitutive trafficking, PI4KB-generated PI4P directs cGAMP-triggered STING Golgi-to-endosome transport via ARMH3 [PMID:36921576], drives non-canonical autophagosome formation downstream of a P38–ULK1 cascade that phosphorylates PI4KB at S256/T263 in oncogenic KRAS contexts [PMID:40055523], and is required for myelination in Schwann cells through Golgi lipid metabolism and protein glycosylation [PMID:33106410]. PI4KB is broadly hijacked by RNA viruses, which redirect it to replication organelles through viral proteins that co-opt ACBD3 or other partners to locally synthesize PI4P [PMID:27009356, PMID:30755512, PMID:31747597].","teleology":[{"year":2001,"claim":"Established PI4KB's core biochemical identity by showing recombinant enzyme produces PI4P with type III kinase kinetics and mapping its phosphorylation-site cluster, framing how the enzyme would later be found to be regulated.","evidence":"Recombinant protein expression, in vitro lipid kinase assay, MALDI-MS and LC-MS/MS phosphosite mapping","pmids":["11277933"],"confidence":"High","gaps":["Did not assign functional consequences to individual phosphosites","No information on physiological kinases responsible"]},{"year":2006,"claim":"Resolved that distinct phosphorylation states partition PI4KB between compartments and tune its activity, linking S294 phosphorylation to Golgi localization/activation and S496/T504 to nuclear speckles and cell viability.","evidence":"Anti-phosphopeptide antibodies, immunofluorescence, microinjection, lipid kinase assay","pmids":["16606619"],"confidence":"Medium","gaps":["Upstream kinases not identified","Nuclear speckle function of PI4KB mechanistically unexplained"]},{"year":2011,"claim":"Defined how PI4KB is spatially targeted by identifying ACBD3 as the Golgi adaptor that recruits it, and revealed this axis is hijacked by viruses to build PI4P-rich replication sites.","evidence":"Co-IP, ACBD3/PI4KB knockdown with viral replication readout, colocalization imaging","pmids":["22124328"],"confidence":"High","gaps":["Structural basis of ACBD3–PI4KB binding not yet defined","Did not address whether recruitment alters catalytic output"]},{"year":2016,"claim":"Provided structural and biochemical mechanism for ACBD3-dependent activation, showing membrane recruitment via ACBD3 boosts PI4KB kinase activity and that viral 3A protein directly activates the kinase, sensitized by ACBD3.","evidence":"NMR and crystal structures, HDX-MS interface mapping, in vitro membrane recruitment and lipid kinase activity assays, mutagenesis","pmids":["27009356","27989622"],"confidence":"High","gaps":["Did not establish the activation mechanism at atomic resolution for the full membrane-bound complex","Relevance of in vitro activation magnitudes to cellular PI4P pools not quantified"]},{"year":2016,"claim":"Validated PI4KB as a druggable antiviral target by defining the ATP-site binding mode of selective inhibitors.","evidence":"X-ray crystallography, enzyme inhibition and antiviral assays","pmids":["28004945"],"confidence":"High","gaps":["Selectivity against the full lipid kinase family not exhaustively profiled","In vivo efficacy not addressed"]},{"year":2017,"claim":"Clarified the post-translational control of PI4KB stability by showing 14-3-3 forms a 2:2 complex upon phosphorylation that protects PI4KB from proteolysis without altering activity or blocking ACBD3 recruitment.","evidence":"SAXS, fluorescence biophysics, proteolysis-protection and activity assays, structural analysis; conserved 14-3-3 recognition shown by crystallography","pmids":["28864297","27827352"],"confidence":"High","gaps":["Cellular consequences of disrupting 14-3-3 binding not tested","Identity of the kinase generating the 14-3-3 site in vivo unresolved"]},{"year":2019,"claim":"Distinguished adaptor-specific recruitment routes by establishing ACBD3 as essential for viral 3A-driven recruitment and characterizing a separate c10orf76–PI4KB complex (via the kinase linker, PKA-modulated) that raises Golgi PI4P and supports Arf1 activation.","evidence":"ACBD3/PI4KB KO and reconstitution with defined mutants, HDX-MS, PI4P measurement, virus replication assays","pmids":["30755512","31829496"],"confidence":"High","gaps":["Whether c10orf76 and ACBD3 pools are physically segregated not yet shown","Role of PKA phosphorylation in physiological signaling undefined"]},{"year":2019,"claim":"Mapped the conformational architecture of PI4KB heterocomplexes, defining stoichiometries (14-3-3:PI4KB:Rab11 of 2:1:1) and membrane-dependent assembly of the ACBD3:PI4KB:Rab11 complex.","evidence":"SAXS, in vitro reconstitution","pmids":["30679637"],"confidence":"Medium","gaps":["Low-resolution flexible models lack atomic detail","Functional output of Rab11-containing complexes not tested"]},{"year":2020,"claim":"Demonstrated a physiological, organ-level requirement for PI4KB by showing Schwann-cell-specific deletion disrupts Golgi morphology, OSBP/GOLPH3 localization, and lipid metabolism, impairing myelination.","evidence":"Conditional PI4KB knockout mice, EM, immunofluorescence, lipid analysis, nerve conduction velocity","pmids":["33106410"],"confidence":"High","gaps":["Did not separate trafficking versus lipid-transport contributions to the myelin defect","Human relevance to peripheral neuropathy not established"]},{"year":2022,"claim":"Identified ARL5A/B GTPases as additional trans-Golgi recruiters of PI4KB that promote PI4P synthesis and secretion.","evidence":"miniTurboID proximity biotinylation, quantitative MS, interaction and functional secretion assays","pmids":["35844135"],"confidence":"Medium","gaps":["PI4KB was one of many hits in a broad Arf-family screen","Direct versus indirect ARL5–PI4KB binding not fully resolved"]},{"year":2023,"claim":"Resolved that distinct adaptor-defined PI4P pools serve different effectors: the C10orf76/ARMH3 distal-Golgi pool feeds CERT-mediated ceramide transport and directs cGAMP-triggered STING trafficking via PI4P-binding AP-1/GGA2.","evidence":"Genome-wide CRISPR screens, super-resolution microscopy, ceramide transport assays, co-IP, RNAi of PI4P-binding proteins, in vivo Armh3 mouse model","pmids":["37195633","36921576"],"confidence":"High","gaps":["How spatial specificity of pools is biophysically maintained not defined","Cross-talk between trafficking and innate-immune pools unexplored"]},{"year":2024,"claim":"Placed PI4KB at the output of a defined GTPase signaling axis, showing ARMH3 is the active-ARL5 effector that activates PI4KB to generate the main TGN PI4P pool controlling GOLPH3 recruitment and glycosylation.","evidence":"Proximity biotinylation, binding assays with active/inactive ARL5 mutants, PI4P measurement, knockdown/knockout functional readouts","pmids":["39580461"],"confidence":"High","gaps":["Direct activation mechanism of PI4KB by ARMH3 not structurally resolved","Quantitative partitioning between ARMH3 and ACBD3 pools not measured"]},{"year":2025,"claim":"Extended PI4KB into signal-driven non-canonical autophagy by showing ULK1 phosphorylates PI4KB at S256/T263 downstream of P38 to drive PI4P production and ATG8ylation in oncogenic KRAS contexts, and identifying SCAMP5 as a TGN recruiter enabling AP-4/ATG9A-dependent presynaptic autophagy.","evidence":"Phosphosite mutagenesis, genetic knockouts, PI4P and autophagy-flux assays, xenograft/KPC mouse models; co-IP and trafficking assays for SCAMP5","pmids":["40055523","40958389"],"confidence":"High","gaps":["SCAMP5 axis rests on single-lab co-IP and knockdown data","How ULK1-driven and Golgi housekeeping PI4P pools are kept distinct unresolved"]},{"year":null,"claim":"It remains unresolved how the cell biophysically segregates the multiple adaptor-defined PI4KB/PI4P pools (ACBD3, ARMH3/ARL5, RAB30, SCAMP5) so that each delivers PI4P to a dedicated effector pathway without cross-interference.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified spatial model integrating all adaptors","Stoichiometry and dynamics of competing complexes in living cells unmeasured","Functional hierarchy among adaptors under physiological signaling undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[9,8,1,2]},{"term_id":"GO:0140097","term_label":"catalytic activity, acting on DNA","supporting_discovery_ids":[9]}],"localization":[{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[1,8,11,13,17]},{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[11,10,13]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[14,18,20]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[12]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1,5,15]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[13,17]}],"complexes":["ACBD3:PI4KB complex","14-3-3:PI4KB (2:2) complex","C10orf76(ARMH3):PI4KB complex","14-3-3:PI4KB:Rab11 complex"],"partners":["ACBD3","C10ORF76/ARMH3","ARL5A","ARL5B","RAB30","SCAMP5","ANXA2","YWHA (14-3-3)"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9UBF8","full_name":"Phosphatidylinositol 4-kinase beta","aliases":["NPIK","PI4K92","PI4KIII"],"length_aa":816,"mass_kda":91.4,"function":"Phosphorylates phosphatidylinositol (PI) in the first committed step in the production of the second messenger inositol-1,4,5,-trisphosphate (PIP). May regulate Golgi disintegration/reorganization during mitosis, possibly via its phosphorylation. Involved in Golgi-to-plasma membrane trafficking (By similarity) (PubMed:10559940, PubMed:11277933, PubMed:12749687, PubMed:9405935). May play an important role in the inner ear development (Microbial infection) Plays an essential role in Aichi virus RNA replication (PubMed:22124328, PubMed:22258260, PubMed:27989622). Recruited by ACBD3 at the viral replication sites (PubMed:22124328, PubMed:27989622) (Microbial infection) Required for cellular spike-mediated entry of human coronavirus SARS-CoV","subcellular_location":"Endomembrane system; Mitochondrion outer membrane; Rough endoplasmic reticulum membrane; Golgi apparatus; Golgi apparatus membrane; Cytoplasm, perinuclear region","url":"https://www.uniprot.org/uniprotkb/Q9UBF8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/PI4KB","classification":"Not Classified","n_dependent_lines":324,"n_total_lines":1208,"dependency_fraction":0.2682119205298013},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000143393","cell_line_id":"CID000142","localizations":[{"compartment":"golgi","grade":3},{"compartment":"cytoplasmic","grade":1},{"compartment":"vesicles","grade":1}],"interactors":[],"url":"https://opencell.sf.czbiohub.org/target/CID000142","total_profiled":1310},"omim":[{"mim_id":"620867","title":"ARMADILLO-LIKE HELICAL DOMAIN-CONTAINING PROTEIN 3; ARMH3","url":"https://www.omim.org/entry/620867"},{"mim_id":"620281","title":"DEAFNESS, AUTOSOMAL DOMINANT 87; DFNA87","url":"https://www.omim.org/entry/620281"},{"mim_id":"616880","title":"TBC1 DOMAIN FAMILY, MEMBER 22B; TBC1D22B","url":"https://www.omim.org/entry/616880"},{"mim_id":"616879","title":"TBC1 DOMAIN FAMILY, MEMBER 22A; TBC1D22A","url":"https://www.omim.org/entry/616879"},{"mim_id":"612374","title":"STIMULATOR OF INTERFERON RESPONSE cGAMP INTERACTOR 1; STING1","url":"https://www.omim.org/entry/612374"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Golgi apparatus","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in 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Interfaces were mapped by hydrogen-deuterium exchange mass spectrometry (HDX-MS).\",\n      \"method\": \"Biochemical reconstitution, crystal structure determination, HDX-MS interface mapping, mutagenesis, lipid kinase activity assay\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure, in vitro reconstitution, mutagenesis, and HDX-MS in a single rigorous study\",\n      \"pmids\": [\"27989622\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PI4KB inhibitors bind in the ATP-binding site of PI4KB; crystallographic analysis confirmed the binding mode and explains their inhibitory activity against PI4KB, with selectivity over other kinases.\",\n      \"method\": \"X-ray crystallography, enzyme inhibition assays, antiviral assays\",\n      \"journal\": \"Journal of medicinal chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structures plus in vitro enzymatic assays, single study but multiple orthogonal methods\",\n      \"pmids\": [\"28004945\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"PI4KB forms a tight 2:2 complex with 14-3-3 proteins upon phosphorylation; 14-3-3 binding does not directly modulate PI4KB enzymatic activity but protects PI4KB from proteolytic degradation in vitro; structural analysis showed 14-3-3 binding does not interfere with ACBD3-mediated membrane recruitment.\",\n      \"method\": \"Biophysical characterization (SAXS, fluorescence methods), proteolysis protection assay, enzymatic activity assay, structural analysis\",\n      \"journal\": \"Journal of structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple orthogonal biophysical methods plus biochemical validation of structural predictions in a single study\",\n      \"pmids\": [\"28864297\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"ACBD3 is an essential mediator of enterovirus 3A-dependent PI4KB recruitment to replication organelles; ACBD3 knockout prevented PI4KB recruitment and impaired 3A Golgi localization; a PI4KB mutant unable to bind ACBD3 failed to restore replication in PI4KB-KO cells; the ACB and CAR domains of ACBD3 are dispensable for PI4KB recruitment.\",\n      \"method\": \"ACBD3 and PI4KB knockout cells, reconstitution with wild-type and mutant proteins, fluorescence microscopy localization, virus replication assays\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple KO cell lines reconstituted with defined mutants, replicated across multiple enterovirus species, multiple orthogonal readouts\",\n      \"pmids\": [\"30755512\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"c10orf76 forms a complex with PI4KB mediated by the kinase linker of PI4KB; PKA-dependent phosphorylation modulates complex formation; PI4KB is required for membrane recruitment of c10orf76 to the Golgi; the intact c10orf76–PI4KB complex increases Golgi PI4P levels and is required for replication of specific enteroviruses; c10orf76 contributes to Arf1 activation at the Golgi.\",\n      \"method\": \"HDX-MS complex characterization, complex-disrupting mutagenesis, PI4P level measurement, virus replication assay, localization studies\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — HDX-MS plus mutagenesis plus functional PI4P and viral replication readouts in a single rigorous study\",\n      \"pmids\": [\"31829496\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PI4KB forms highly flexible heterocomplexes including ACBD3, 14-3-3, and Rab11 proteins; the 14-3-3:PI4KB:Rab11 complex has 2:1:1 stoichiometry; the ACBD3:PI4KB complex exists in very compact and extended conformations; membrane is necessary for formation of the ACBD3:PI4KB:Rab11 complex at physiological concentrations.\",\n      \"method\": \"Small angle X-ray scattering (SAXS), in vitro reconstitution\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — SAXS structural characterization and in vitro reconstitution, single lab, single study\",\n      \"pmids\": [\"30679637\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"PI4K92 (PI4KB) phosphorylated at Ser-294 localizes exclusively to the Golgi and this phosphorylation increases lipid kinase activity; PI4K92 phosphorylated at Ser-496 and Thr-504 localizes to nuclear speckles dynamically and is required for cell viability (microinjection of anti-pSer-496 antibody caused apoptosis).\",\n      \"method\": \"Anti-phosphopeptide antibodies, indirect immunofluorescence, microinjection, lipid kinase activity assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct subcellular localization with site-specific antibodies plus functional readout (kinase activity, microinjection phenotype), single lab\",\n      \"pmids\": [\"16606619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Recombinant PI4K92 (PI4KB) produces PtdIns4P as product; is characterized as a type III PI4K (high Km for ATP and PtdIns in millimolar range, IC50 ~300 nM for Wortmannin); multiple phosphorylation sites mapped by MALDI-MS and LC-MS/MS at S258, T263, S266, S277, S294, T423, S496, T504 within a designated phosphorylation domain.\",\n      \"method\": \"Recombinant protein expression, in vitro lipid kinase assay, MALDI-MS, LC-MS/MS phosphorylation site mapping\",\n      \"journal\": \"European journal of biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro enzymatic characterization plus mass spectrometry phosphorylation site mapping, directly on recombinant PI4KB\",\n      \"pmids\": [\"11277933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ARL5A and ARL5B GTPases interact with PI4KB at the trans-Golgi and recruit it, thereby promoting PI4KB's function in PI4P synthesis and protein secretion.\",\n      \"method\": \"Proximity biotinylation (miniTurboID), quantitative mass spectrometry, protein interaction assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proximity labeling plus functional PI4P and secretion readouts, but PI4KB is one of many interactors identified in a broader Arf-family screen\",\n      \"pmids\": [\"35844135\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ARMH3 (C10orf76) is an effector of active ARL5 at the TGN; ARMH3 activates PI4KB to account for the main pool of PI4P at the TGN; this contributes to GOLPH3 recruitment and glycan modifications; the SYS1-ARFRP1-ARL5-ARMH3 axis regulates PI4KB-dependent PI4P generation at the TGN.\",\n      \"method\": \"Proximity biotinylation, protein interaction assays (active vs. inactive ARL5 mutants), PI4P level measurement, knockdown/knockout with functional readouts (GOLPH3 localization, glycan modification)\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (proximity labeling, direct binding assays with GTPase mutants, functional PI4P and downstream readouts), rigorous study\",\n      \"pmids\": [\"39580461\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"ARMH3 interacts with STING at the Golgi upon cGAMP stimulation and recruits PI4KB to synthesize PI4P, which directs STING Golgi-to-endosome trafficking via PI4P-binding proteins AP-1 and GGA2; disrupting PI4P-dependent transport impaired STING activation; elevated cellular PI4P was sufficient for cGAS-independent STING activation.\",\n      \"method\": \"Genome-wide CRISPR-Cas9 screen, co-immunoprecipitation, PI4P measurement, RNAi of PI4P-binding proteins, in vivo mouse model (Armh3fl/fl LyzCre/Cre)\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide unbiased screen plus mechanistic validation with co-IP, multiple RNAi knockdowns, PI4P manipulation, and in vivo mouse model\",\n      \"pmids\": [\"36921576\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"The C10orf76–PI4KB axis generates a specific pool of PI4P at distal Golgi subregions; CERT preferentially utilizes PI4P generated by PI4KB recruited via C10orf76 (not ACBD3) for ER-to-Golgi ceramide trafficking; C10orf76 localizes predominantly at distal Golgi where sphingomyelin synthesis occurs, while ACBD3 is at more proximal regions.\",\n      \"method\": \"Genome-wide screening, PI4P measurement, super-resolution microscopy, ceramide transport assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide unbiased screen plus super-resolution microscopy plus lipid transfer functional assay, multiple orthogonal methods\",\n      \"pmids\": [\"37195633\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RAB30 interacts with PI4KB and recruits it to the Golgi apparatus and GAS-containing autophagosome-like vacuoles (GcAVs); PI4KB knockout suppressed autophagy by inhibiting GcAV formation, resulting in increased GAS survival; PtdIns4P is crucial for GcAV formation.\",\n      \"method\": \"Co-immunoprecipitation (RAB30–PI4KB interaction), PI4KB knockout cells, autophagy/GcAV formation assay, PI4P depletion/repletion experiments\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP interaction plus KO phenotype plus PI4P functional rescue, single lab\",\n      \"pmids\": [\"30290718\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PI4KB is recruited to inclusion bodies formed by ER membrane remodeling during HPIV3 infection; the HPIV3 phosphoprotein (P) recruits PI4KB to IBs to generate PI4P, creating a PI4P-enriched microenvironment that promotes viral replication; HRSV nucleoprotein similarly recruits PI4KB to IBs.\",\n      \"method\": \"Co-immunoprecipitation, PI4P measurement, colocalization imaging, knockdown/loss-of-function viral replication assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, PI4P measurement, and functional replication readouts, single lab\",\n      \"pmids\": [\"31747597\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Under PI4KB inhibition, PI4KB activity has distinct functions in both proteolytic processing of the enteroviral polyprotein and in replication organelle (RO) biogenesis; a PI4KB-inhibitor-resistant escape mutation corrects a proteolytic processing defect, revealing an unexpected role of PI4KB in viral polyprotein processing.\",\n      \"method\": \"PI4KB inhibitor-resistant mutant virus selection, electron tomography of ROs, viral polyprotein processing analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — resistance mutation mapping plus structural (EM) and biochemical readouts, single lab\",\n      \"pmids\": [\"29045829\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"SC-specific genetic inactivation of PI4KB in mice disrupts Golgi morphology in Schwann cells, causes disappearance of OSBP at cis-Golgi and loss of GOLPH3 from the entire Golgi, reduces cholesterol and sphingomyelin in sciatic nerves, reduces caveolae, and results in thinner myelin, abnormal nodes of Ranvier, and impaired engulfment of nerve fibers; demonstrating PI4KB Golgi function is required for myelination through lipid metabolism and protein glycosylation control.\",\n      \"method\": \"Conditional PI4KB knockout mice (SC-specific), electron microscopy, immunofluorescence, lipid analysis, nerve conduction velocity measurement\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — conditional KO mouse model with multiple orthogonal cellular, biochemical, and physiological readouts\",\n      \"pmids\": [\"33106410\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Oncogenic KRAS induces a non-canonical autophagy pathway (RINCAA) involving a P38-ULK1-PI4KB-WIPI2 signaling cascade; ULK1 phosphorylates PI4KB at S256 and T263, triggering PI4P production, ATG8ylation, and non-canonical autophagosome formation; inhibiting these phosphorylation sites reduces RINCAA activity and tumor growth in xenograft and KPC pancreatic cancer models.\",\n      \"method\": \"Genetic knockouts, phosphorylation site mutagenesis, PI4P measurement, autophagy flux assays, in vivo xenograft and KPC mouse models\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mutagenesis of specific phosphorylation sites plus multiple in vivo cancer models plus multiple orthogonal autophagy readouts\",\n      \"pmids\": [\"40055523\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ANXA2 interacts with both EV71 3D polymerase and PI4KB; the annexin domain of ANXA2 mediates 3D binding; ANXA2 localizes to replication organelles; ANXA2 overexpression stimulates PI4P production and promotes the PI4KB–3D interaction, facilitating formation of a higher-order ANXA2–PI4KB–3D protein complex at the viral replication site.\",\n      \"method\": \"Co-immunoprecipitation, ANXA2 knockout, PI4P level measurement, colocalization imaging\",\n      \"journal\": \"Virologica Sinica\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, KO with PI4P measurement, colocalization, single lab\",\n      \"pmids\": [\"34196914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SCAMP5 is a binding partner of PI4KB/PI4KIIIβ at the TGN; SCAMP5 controls PI4KB recruitment to the TGN and subsequent PI4P production; PI4P is essential for AP-4 recruitment; SCAMP5 depletion disrupts AP-4-mediated ATG9A trafficking to presynaptic sites, impairing presynaptic autophagy.\",\n      \"method\": \"Co-immunoprecipitation (SCAMP5–PI4KB interaction), SCAMP5 knockdown with PI4P measurement, AP-4 localization, ATG9A trafficking assay, autophagosome formation assay\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus functional PI4P and trafficking readouts, single lab\",\n      \"pmids\": [\"40958389\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HCV NS5A competes with PI4KB for binding to ACBD3 in a genotype-dependent manner; ACBD3 binds GT1b NS5A with higher affinity than GT2a NS5A, correlating with higher PI4KB/PI4P colocalization in GT1b-infected cells; NS5A displaces PI4KB from ACBD3 to allow PI4KB relocation to replication sites.\",\n      \"method\": \"Co-immunoprecipitation (competitive binding), colocalization imaging\",\n      \"journal\": \"Antiviral research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP competition assay plus colocalization, single lab, two orthogonal approaches\",\n      \"pmids\": [\"24792752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"A yeast 14-3-3 protein crystal structure bound to a phosphopeptide derived from human PI4KB was determined, demonstrating that the 14-3-3 recognition mode for PI4KB phosphopeptide is highly evolutionarily conserved.\",\n      \"method\": \"X-ray crystallography of 14-3-3 bound to PI4KB-derived phosphopeptide\",\n      \"journal\": \"Acta crystallographica Section F\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — crystal structure with PI4KB peptide, single study, structural only without functional mutagenesis of PI4KB itself\",\n      \"pmids\": [\"27827352\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"PI4KB (PI4K IIIβ) is a Golgi/TGN-localized lipid kinase that phosphorylates phosphatidylinositol to produce PI4P; it is recruited to the Golgi by multiple adaptors (ACBD3, C10orf76/ARMH3, RAB30, ARL5A/B, SCAMP5), regulated by 14-3-3 protein binding (which stabilizes but does not directly alter its activity) and by phosphorylation at multiple sites (including PKD-mediated and ULK1-mediated sites), and generates distinct PI4P pools that control membrane trafficking, ceramide transport, Golgi function, myelination, STING antiviral signaling, and non-canonical autophagy; PI4KB is also hijacked by numerous RNA viruses through viral proteins that co-opt ACBD3 or other adaptors to redirect PI4KB to viral replication organelles.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"PI4KB (PI4KIIIβ) is a type III phosphatidylinositol 4-kinase that phosphorylates phosphatidylinositol to generate PI4P, and the Golgi/trans-Golgi PI4P pools it produces control membrane trafficking, secretion, lipid transport, antiviral signaling, autophagy, and myelination [#9, #11, #17]. Catalytically, PI4KB is a high-Km type III kinase whose activity is tuned both by membrane recruitment and by phosphorylation at a defined cluster of sites (S294 increasing kinase activity, with additional sites including S496/T504 governing localization) [#8, #9]. Its function depends on recruitment to specific membranes by a panel of adaptors: ACBD3 binds PI4KB and, upon membrane recruitment, stimulates its lipid kinase activity to support Golgi function [#1, #2], while ARL5A/B GTPases and their effector ARMH3/C10orf76 define a SYS1–ARFRP1–ARL5–ARMH3 axis that generates the main TGN PI4P pool driving GOLPH3 recruitment, glycosylation, and secretion [#6, #10, #11]; RAB30 and SCAMP5 provide additional recruitment routes [#14, #20]. PI4KB partitions into distinct subcellular PI4P pools, with the C10orf76/ARMH3-recruited pool at distal Golgi feeding CERT-mediated ceramide transport, separate from the ACBD3-associated proximal pool [#13]. PI4KB is regulated post-translationally by phosphorylation-dependent 14-3-3 binding, which forms a 2:2 complex that protects PI4KB from proteolysis without altering its catalytic activity or blocking ACBD3-mediated recruitment [#4, #7]. Beyond constitutive trafficking, PI4KB-generated PI4P directs cGAMP-triggered STING Golgi-to-endosome transport via ARMH3 [#12], drives non-canonical autophagosome formation downstream of a P38–ULK1 cascade that phosphorylates PI4KB at S256/T263 in oncogenic KRAS contexts [#18], and is required for myelination in Schwann cells through Golgi lipid metabolism and protein glycosylation [#17]. PI4KB is broadly hijacked by RNA viruses, which redirect it to replication organelles through viral proteins that co-opt ACBD3 or other partners to locally synthesize PI4P [#1, #5, #15].\",\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Established PI4KB's core biochemical identity by showing recombinant enzyme produces PI4P with type III kinase kinetics and mapping its phosphorylation-site cluster, framing how the enzyme would later be found to be regulated.\",\n      \"evidence\": \"Recombinant protein expression, in vitro lipid kinase assay, MALDI-MS and LC-MS/MS phosphosite mapping\",\n      \"pmids\": [\"11277933\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not assign functional consequences to individual phosphosites\", \"No information on physiological kinases responsible\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Resolved that distinct phosphorylation states partition PI4KB between compartments and tune its activity, linking S294 phosphorylation to Golgi localization/activation and S496/T504 to nuclear speckles and cell viability.\",\n      \"evidence\": \"Anti-phosphopeptide antibodies, immunofluorescence, microinjection, lipid kinase assay\",\n      \"pmids\": [\"16606619\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Upstream kinases not identified\", \"Nuclear speckle function of PI4KB mechanistically unexplained\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined how PI4KB is spatially targeted by identifying ACBD3 as the Golgi adaptor that recruits it, and revealed this axis is hijacked by viruses to build PI4P-rich replication sites.\",\n      \"evidence\": \"Co-IP, ACBD3/PI4KB knockdown with viral replication readout, colocalization imaging\",\n      \"pmids\": [\"22124328\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of ACBD3–PI4KB binding not yet defined\", \"Did not address whether recruitment alters catalytic output\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Provided structural and biochemical mechanism for ACBD3-dependent activation, showing membrane recruitment via ACBD3 boosts PI4KB kinase activity and that viral 3A protein directly activates the kinase, sensitized by ACBD3.\",\n      \"evidence\": \"NMR and crystal structures, HDX-MS interface mapping, in vitro membrane recruitment and lipid kinase activity assays, mutagenesis\",\n      \"pmids\": [\"27009356\", \"27989622\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish the activation mechanism at atomic resolution for the full membrane-bound complex\", \"Relevance of in vitro activation magnitudes to cellular PI4P pools not quantified\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Validated PI4KB as a druggable antiviral target by defining the ATP-site binding mode of selective inhibitors.\",\n      \"evidence\": \"X-ray crystallography, enzyme inhibition and antiviral assays\",\n      \"pmids\": [\"28004945\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Selectivity against the full lipid kinase family not exhaustively profiled\", \"In vivo efficacy not addressed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Clarified the post-translational control of PI4KB stability by showing 14-3-3 forms a 2:2 complex upon phosphorylation that protects PI4KB from proteolysis without altering activity or blocking ACBD3 recruitment.\",\n      \"evidence\": \"SAXS, fluorescence biophysics, proteolysis-protection and activity assays, structural analysis; conserved 14-3-3 recognition shown by crystallography\",\n      \"pmids\": [\"28864297\", \"27827352\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular consequences of disrupting 14-3-3 binding not tested\", \"Identity of the kinase generating the 14-3-3 site in vivo unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Distinguished adaptor-specific recruitment routes by establishing ACBD3 as essential for viral 3A-driven recruitment and characterizing a separate c10orf76–PI4KB complex (via the kinase linker, PKA-modulated) that raises Golgi PI4P and supports Arf1 activation.\",\n      \"evidence\": \"ACBD3/PI4KB KO and reconstitution with defined mutants, HDX-MS, PI4P measurement, virus replication assays\",\n      \"pmids\": [\"30755512\", \"31829496\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether c10orf76 and ACBD3 pools are physically segregated not yet shown\", \"Role of PKA phosphorylation in physiological signaling undefined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Mapped the conformational architecture of PI4KB heterocomplexes, defining stoichiometries (14-3-3:PI4KB:Rab11 of 2:1:1) and membrane-dependent assembly of the ACBD3:PI4KB:Rab11 complex.\",\n      \"evidence\": \"SAXS, in vitro reconstitution\",\n      \"pmids\": [\"30679637\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Low-resolution flexible models lack atomic detail\", \"Functional output of Rab11-containing complexes not tested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Demonstrated a physiological, organ-level requirement for PI4KB by showing Schwann-cell-specific deletion disrupts Golgi morphology, OSBP/GOLPH3 localization, and lipid metabolism, impairing myelination.\",\n      \"evidence\": \"Conditional PI4KB knockout mice, EM, immunofluorescence, lipid analysis, nerve conduction velocity\",\n      \"pmids\": [\"33106410\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not separate trafficking versus lipid-transport contributions to the myelin defect\", \"Human relevance to peripheral neuropathy not established\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified ARL5A/B GTPases as additional trans-Golgi recruiters of PI4KB that promote PI4P synthesis and secretion.\",\n      \"evidence\": \"miniTurboID proximity biotinylation, quantitative MS, interaction and functional secretion assays\",\n      \"pmids\": [\"35844135\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"PI4KB was one of many hits in a broad Arf-family screen\", \"Direct versus indirect ARL5–PI4KB binding not fully resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved that distinct adaptor-defined PI4P pools serve different effectors: the C10orf76/ARMH3 distal-Golgi pool feeds CERT-mediated ceramide transport and directs cGAMP-triggered STING trafficking via PI4P-binding AP-1/GGA2.\",\n      \"evidence\": \"Genome-wide CRISPR screens, super-resolution microscopy, ceramide transport assays, co-IP, RNAi of PI4P-binding proteins, in vivo Armh3 mouse model\",\n      \"pmids\": [\"37195633\", \"36921576\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How spatial specificity of pools is biophysically maintained not defined\", \"Cross-talk between trafficking and innate-immune pools unexplored\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed PI4KB at the output of a defined GTPase signaling axis, showing ARMH3 is the active-ARL5 effector that activates PI4KB to generate the main TGN PI4P pool controlling GOLPH3 recruitment and glycosylation.\",\n      \"evidence\": \"Proximity biotinylation, binding assays with active/inactive ARL5 mutants, PI4P measurement, knockdown/knockout functional readouts\",\n      \"pmids\": [\"39580461\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct activation mechanism of PI4KB by ARMH3 not structurally resolved\", \"Quantitative partitioning between ARMH3 and ACBD3 pools not measured\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended PI4KB into signal-driven non-canonical autophagy by showing ULK1 phosphorylates PI4KB at S256/T263 downstream of P38 to drive PI4P production and ATG8ylation in oncogenic KRAS contexts, and identifying SCAMP5 as a TGN recruiter enabling AP-4/ATG9A-dependent presynaptic autophagy.\",\n      \"evidence\": \"Phosphosite mutagenesis, genetic knockouts, PI4P and autophagy-flux assays, xenograft/KPC mouse models; co-IP and trafficking assays for SCAMP5\",\n      \"pmids\": [\"40055523\", \"40958389\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"SCAMP5 axis rests on single-lab co-IP and knockdown data\", \"How ULK1-driven and Golgi housekeeping PI4P pools are kept distinct unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how the cell biophysically segregates the multiple adaptor-defined PI4KB/PI4P pools (ACBD3, ARMH3/ARL5, RAB30, SCAMP5) so that each delivers PI4P to a dedicated effector pathway without cross-interference.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified spatial model integrating all adaptors\", \"Stoichiometry and dynamics of competing complexes in living cells unmeasured\", \"Functional hierarchy among adaptors under physiological signaling undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [9, 8, 1, 2]},\n      {\"term_id\": \"GO:0140097\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"GO:0016301\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [1, 8, 11, 13, 17]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [11, 10, 13]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [14, 18, 20]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [12]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 5, 15]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [13, 17]}\n    ],\n    \"complexes\": [\n      \"ACBD3:PI4KB complex\",\n      \"14-3-3:PI4KB (2:2) complex\",\n      \"C10orf76(ARMH3):PI4KB complex\",\n      \"14-3-3:PI4KB:Rab11 complex\"\n    ],\n    \"partners\": [\n      \"ACBD3\",\n      \"C10orf76/ARMH3\",\n      \"ARL5A\",\n      \"ARL5B\",\n      \"RAB30\",\n      \"SCAMP5\",\n      \"ANXA2\",\n      \"YWHA (14-3-3)\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}