{"gene":"OLA1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2007,"finding":"Human OLA1 (hOLA1) binds and hydrolyzes ATP more efficiently than GTP, establishing it as an ATPase rather than a GTPase within the Obg/YchF subfamily. X-ray crystal structure of hOLA1 bound to the non-hydrolyzable ATP analogue AMPPCP explained the altered nucleotide specificity compared to other Obg-family GTPases.","method":"Biochemical nucleotide-binding and hydrolysis assays; X-ray crystallography with AMPPCP-bound structure","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus in vitro enzymatic characterization, replicated across multiple YchF orthologs in the same study","pmids":["17430889"],"is_preprint":false},{"year":2009,"finding":"OLA1 functions as a negative regulator of the cellular antioxidant response through nontranscriptional mechanisms. Knockdown increased resistance to oxidizing agents (tBH, diamide) and reduced intracellular ROS and glutathione depletion without altering antioxidant gene mRNA levels or requiring de novo protein synthesis.","method":"RNAi knockdown; OLA1 overexpression; cell viability and ROS assays; cycloheximide block experiments; qRT-PCR of antioxidant genes","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — bidirectional manipulation (KD and OE), multiple orthogonal readouts, mechanistic dissection ruling out transcriptional mechanism","pmids":["19706404"],"is_preprint":false},{"year":2009,"finding":"Knockdown of OLA1 inhibits breast cancer cell migration and invasion through modulation of intracellular ROS levels, linking OLA1's regulation of ROS to cytoskeletal motility.","method":"siRNA knockdown; wound-healing and transwell invasion assays; ROS measurement; N-acetylcysteine treatment","journal":"Journal of Zhejiang University. Science. B","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, multiple functional assays but no direct molecular partner identified for motility phenotype","pmids":["19882753"],"is_preprint":false},{"year":2013,"finding":"OLA1 directly binds to the amino-terminal region of BRCA1 and to γ-tubulin; it interacts with the carboxy-terminal region of BARD1. OLA1 localizes to centrosomes in interphase and to the spindle pole in mitosis. OLA1 knockdown causes centrosome amplification and activation of microtubule aster formation. A cancer-derived mutation E168Q abrogates BRCA1 binding and fails to rescue centrosome amplification.","method":"Mass spectrometry identification; co-immunoprecipitation; direct binding assays; immunofluorescence localization; RNAi knockdown; rescue with mutant constructs","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, direct binding, localization with functional consequence, mutagenesis rescue experiments","pmids":["24289923"],"is_preprint":false},{"year":2013,"finding":"OLA1 stabilizes HSP70 by binding to its carboxyl-terminus variable domain, thereby blocking recruitment of the E3 ubiquitin ligase CHIP and preventing CHIP-mediated ubiquitination and degradation of HSP70. OLA1 downregulation reduces steady-state HSP70 levels and impairs heat-shock-induced HSP70 induction, increasing cellular sensitivity to heat shock.","method":"RNAi knockdown; targeted gene disruption; OLA1 overexpression; protein-protein interaction (co-IP/pulldown); ubiquitination assay; thermal resistance assay","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 2 / Moderate — bidirectional manipulation, direct interaction assay, ubiquitination mechanistic assay, functional thermotolerance readout in single lab","pmids":["23412384"],"is_preprint":false},{"year":2014,"finding":"OLA1 negatively regulates cell-matrix adhesion and spreading. OLA1-deficient cells have elevated FAK protein levels and decreased Ser3 phosphorylation of cofilin, while OLA1-overexpressing cells show opposite changes, indicating OLA1 modulates adhesion at least partly through FAK expression and cofilin phosphorylation.","method":"RNAi knockdown; OLA1 overexpression; cell adhesion/spreading assays; western blot for FAK and p-cofilin","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — bidirectional manipulation with two molecular readouts, single lab, no direct interaction between OLA1 and FAK/cofilin demonstrated","pmids":["24486488"],"is_preprint":false},{"year":2015,"finding":"OLA1 inhibits protein synthesis and promotes the integrated stress response (ISR) by binding eIF2, hydrolyzing GTP in that context, and interfering with ternary complex (eIF2-GTP-tRNAi) formation. OLA1 depletion causes hypoactive ISR, reduces CHOP induction, and promotes tumor growth and metastasis in vivo.","method":"Co-immunoprecipitation (OLA1-eIF2 interaction); GTPase activity assay; ternary complex formation assay; polysome profiling; RNAi knockdown; xenograft tumor models","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct binding to eIF2, enzymatic activity assay, TC formation assay, in vivo xenograft validation; multiple orthogonal methods in single lab","pmids":["26283179"],"is_preprint":false},{"year":2016,"finding":"OLA1 is a GSK3β-interacting protein and inhibits GSK3β activity by mediating its Ser9 phosphorylation, thereby suppressing GSK3β-mediated degradation of Snail, which promotes E-cadherin downregulation and contributes to TGF-β-induced epithelial-mesenchymal transition (EMT).","method":"Co-immunoprecipitation (OLA1-GSK3β); western blot for GSK3β Ser9 phosphorylation, Snail, and E-cadherin; RNAi knockdown; EMT assays with TGF-β treatment","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP plus phosphorylation readout and EMT functional assays, single lab","pmids":["26863455"],"is_preprint":false},{"year":2016,"finding":"OLA1 is required for normal mammalian development. Ola1-knockout mouse embryos have growth retardation and developmental delay. Primary Ola1-/- MEFs show impaired proliferation due to defective cell-cycle progression, with reduced cyclins D1/E1, attenuated Rb phosphorylation, and elevated p21 protein. p21 accumulation is due to enhanced mRNA translation that is reversed by eIF2α dephosphorylation inhibitor, placing OLA1 upstream of eIF2-mediated p21 translational control.","method":"Ola1 knockout mice; primary MEF culture; BrdU/cell-cycle analysis; western blot; polysome/translation assays; eIF2α pharmacological manipulation; p21-/-/Ola1-/- double-knockout rescue","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic knockout, double-knockout epistasis, pharmacological rescue, multiple molecular and cellular readouts","pmids":["27481995"],"is_preprint":false},{"year":2018,"finding":"OLA1 requires its interaction with BARD1 to properly regulate centrosome number. Three OLA1 missense mutants that fail to bind BARD1 are deficient in centrosome number regulation. Phosphomimetic mutations at specific OLA1 residues restore BARD1 binding and rescue centrosome amplification. BARD1 mutant V695L (cancer-associated) fails to bind OLA1 and cannot rescue BARD1 knockdown-induced centrosome amplification.","method":"Co-immunoprecipitation; site-directed mutagenesis of OLA1 (phosphorylation, acetylation, ATP-binding residues); overexpression of mutant constructs; RNAi knockdown/rescue; centrosome counting by immunofluorescence","journal":"Molecular cancer research : MCR","confidence":"High","confidence_rationale":"Tier 2 / Moderate — systematic mutagenesis, binding assays, functional rescue experiments, bidirectional manipulation of BARD1","pmids":["29858377"],"is_preprint":false},{"year":2019,"finding":"OLA1 is N-terminally methylated in vivo by the N-terminal methyltransferase NTMT1, as demonstrated by activity-based substrate profiling and validation in NTMT1 knockout cells.","method":"Activity-based substrate profiling with Hey-SAM analogue; CRISPR-Cas9 NTMT1 knockout HEK293FT cells; mass spectrometry validation","journal":"Chemical science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chemical biology profiling plus genetic KO validation, single lab","pmids":["31857877"],"is_preprint":false},{"year":2019,"finding":"Decreased OLA1 expression in PPHN enhances CHIP affinity for the Hsp70-SOD2 complex, facilitating SOD2 ubiquitination and proteasomal degradation. OLA1 acts as a molecular chaperone whose stress-induced activity prevents CHIP-mediated SOD2 degradation; ola1-/- mice recapitulate PPHN phenotypes including SOD2 downregulation and pulmonary vascular remodeling.","method":"Patient/fetal lamb tissue analysis; OLA1 KO mice; co-immunoprecipitation of CHIP-Hsp70-SOD2; ubiquitination assays; right ventricular pressure measurement","journal":"Hypertension (Dallas, Tex. : 1979)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO in vivo, interaction assays, disease model; single lab","pmids":["31476900"],"is_preprint":false},{"year":2020,"finding":"OLA1 localizes to meiotic spindles in mouse oocytes (co-localizing with spindle structures after GVBD, confirmed by nocodazole treatment). OLA1 knockdown results in abnormal/multipolar spindle assembly, premature anaphase onset due to precocious spindle assembly checkpoint (SAC) inactivation, and impaired germinal vesicle breakdown.","method":"Immunofluorescence/confocal microscopy; nocodazole treatment; siRNA microinjection; chromosome spreading; polar body extrusion assays","journal":"PeerJ","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — localization with functional consequence, RNAi knockdown with multiple readouts, single lab","pmids":["31915569"],"is_preprint":false},{"year":2020,"finding":"HIV p17 interacts with OLA1 and disrupts the OLA1-GSK3β complex, leading to GSK3β hyperactivation, suppression of autophagy, and enhanced proliferation of HIV-infected T cells under glucose starvation conditions.","method":"Co-immunoprecipitation (p17-OLA1, OLA1-GSK3β); autophagy flux assays; T cell proliferation assays under glucose starvation","journal":"Journal of medical virology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP interaction plus functional autophagy and proliferation readouts, single lab","pmids":["32790080"],"is_preprint":false},{"year":2021,"finding":"ZFAS1 lncRNA recognizes the OBG-type functional domain of OLA1, facilitating exposure of its ATP-binding site (NVGKST, residues 32–37), enhancing OLA1 protein ATPase activity, and accelerating ATP hydrolysis and the Warburg effect in colorectal cancer cells.","method":"RNA pulldown; RIP assay; ATP hydrolysis assay; ECAR/glycolysis assay; mutagenesis of OLA1 ATP-binding site","journal":"Journal of hematology & oncology","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — RNA-protein interaction assay plus enzymatic activity measurement, single lab","pmids":["34743750"],"is_preprint":false},{"year":2022,"finding":"BARD1 acts as an ATPase activating protein (AAP) for OLA1. The BARD1 BRCT domain binds the OLA1 TGS domain via a conserved BUDR motif and allosterically increases OLA1 ATPase turnover number (kcat). Cancer-associated BARD1 mutation V695L reduces OLA1 binding and activation, as revealed by a 1.88 Å crystal structure of the V695L BRCT mutant.","method":"Enzyme kinetics assays; X-ray crystallography (BRCT V695L mutant at 1.88 Å); biophysical/biochemical binding assays; mutagenesis","journal":"Biochimica et biophysica acta. General subjects","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure plus enzyme kinetics plus mutagenesis, multiple orthogonal methods in single study","pmids":["35134491"],"is_preprint":false},{"year":2022,"finding":"OLA1 knockout in colorectal cancer cells activates GSK3β and downregulates HIF1α and its target CA9 at the mRNA/protein level, linking OLA1 to the HIF1α/CA9 hypoxic signaling axis through GSK3β.","method":"CRISPR-Cas9 OLA1 knockout; mRNA sequencing; western blot for GSK3β, HIF1α, CA9; xenograft tumor models","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — genetic KO with pathway readouts, single lab, no direct OLA1-HIF1α interaction demonstrated","pmids":["35440019"],"is_preprint":false},{"year":2023,"finding":"Aurora A binds OLA1 and polyubiquitinates it, targeting OLA1 for proteasomal degradation. NEK2 phosphorylates OLA1 at T124, which increases OLA1 binding to Aurora A and enhances Aurora A-mediated polyubiquitination. The kinase activity of Aurora A suppresses its own E3 ligase activity toward OLA1. Reduction of centrosomal OLA1 by this mechanism promotes pericentriolar material protein recruitment in G2 phase, required for centrosome maturation.","method":"Co-immunoprecipitation; in vitro ubiquitination assay; mutagenesis (T124 phosphorylation site); proteasome inhibitor experiments; immunofluorescence of centrosomal OLA1; overexpression rescue","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro ubiquitination reconstitution, mutagenesis, in vivo degradation assays, centrosome functional readout; single lab with multiple orthogonal methods","pmids":["37481721"],"is_preprint":false},{"year":2023,"finding":"OLA1 phosphorylation at Ser232/Tyr236 triggers its translocation from cytoplasm/mitochondria into the nucleus. Subsequent phosphorylation at Thr325 switches its biochemical activity from ATPase to GTPase and promotes expression of nuclear-encoded mitochondrial bioenergetic genes. ERK1/2 drives this process and is restrained by PP1A. OLA1 T325A (phosphoresistant) mutant blocks nuclear translocation and compromises mitochondrial gene expression. OLA1 knockout mice have fewer mitochondria, lower ATP, and higher lactate.","method":"Phospho-site mutagenesis; subcellular fractionation; immunofluorescence; ATPase/GTPase activity assays with phosphomimetic mutants; ERK inhibition; PP1A manipulation; OLA1 KO mice; metabolite measurements","journal":"American journal of respiratory cell and molecular biology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — mutagenesis of phosphorylation sites, enzymatic activity switch demonstrated, KO mice with metabolic phenotype, multiple orthogonal methods","pmids":["36481055"],"is_preprint":false},{"year":2024,"finding":"HIV-1 p17 (but not HIV-2 or SIV p17) binds OLA1 and inhibits OLA1's interaction with STING, thereby blocking OLA1-mediated suppression of STING translocation and phosphorylation. OLA1 normally interacts with STING and inhibits STING activation upon cGAMP stimulation. HIV-1 p17 also specifically promotes OLA1 ATPase and GTPase activities.","method":"Co-immunoprecipitation (OLA1-STING, p17-OLA1); cGAMP stimulation assays; STING translocation/phosphorylation assays; comparative HIV-1 vs HIV-2/SIV p17 experiments; ATPase/GTPase activity assays","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP plus functional STING signaling assays, comparative species analysis; single lab","pmids":["38132845"],"is_preprint":false},{"year":2025,"finding":"In fission yeast (S. pombe), Ola1 physically interacts with MAPK/Pmk1 and its upstream kinase Pek1 (MAPKK), inhibiting MAPK/Pmk1 signaling to prevent excessive mitochondrial ROS accumulation. Absence of Ola1 increases mtROS, promotes nuclear localization of Hsf1, and upregulates Ssa1 (Hsp70 homolog).","method":"Co-immunoprecipitation (Ola1-Pmk1, Ola1-Pek1); mitochondrial ROS measurements; ola1 deletion; Hsf1 localization assay; western blot","journal":"Microbiological research","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP with functional mtROS readout in a yeast ortholog model; single lab","pmids":["40543417"],"is_preprint":false},{"year":2025,"finding":"OLA1 interacts with Keap1 and, when STING is activated, enhanced STING-OLA1 interaction disrupts the OLA1-Keap1 complex, liberating Keap1 to promote Nrf2 degradation and ferroptosis in granulosa cells. This STING-OLA1-Keap1-Nrf2 axis is mechanistically linked to premature ovarian failure.","method":"Co-immunoprecipitation (STING-OLA1, OLA1-Keap1); siRNA knockdown; Nrf2 protein stability assays; ferroptosis markers; murine POF model; molecular docking (Icariin-STING)","journal":"International journal of biological macromolecules","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP interactions with functional Nrf2/ferroptosis readouts in cellular and in vivo models; single lab","pmids":["41352507"],"is_preprint":false},{"year":2023,"finding":"Vitexin binds OLA1 (identified by tissue thermal proteome profiling and molecular docking) and the OLA1-vitexin complex interacts with Keap1, disrupting the Keap1-Nrf2 interaction and activating Nrf2. siRNA knockdown of OLA1 in Caco-2 cells confirmed OLA1's role in mediating Nrf2 protein expression and anti-inflammatory effects.","method":"Tissue thermal proteome profiling; molecular docking; siRNA knockdown; Nrf2 protein level assays; inflammatory cytokine measurements","journal":"Journal of agricultural and food chemistry","confidence":"Low","confidence_rationale":"Tier 3–4 / Weak — computational docking plus single siRNA KD experiment; no direct biochemical reconstitution of OLA1-Keap1 interaction","pmids":["37856434"],"is_preprint":false},{"year":2026,"finding":"Bi-allelic loss-of-function variants in OLA1 cause a human neurodevelopmental disorder with joint hypermobility. Patient-derived fibroblasts recapitulate impaired migration and proliferation. Neurons derived from proband fibroblasts show impaired adhesion and cytoskeletal control. In C. elegans, ola-1 deficiency reduces neurite numbers and suppresses microtubule dynamics and axon regrowth, placing OLA1 in a pathway regulating cytoskeletal dynamics through FAK levels.","method":"Sanger/exome sequencing in 14 individuals from 9 families; proband-derived fibroblast migration/proliferation assays; iPSC-derived neuron adhesion/cytoskeletal assays; C. elegans ola-1 knockout; transcriptomics","journal":"American journal of human genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — patient genetic evidence plus proband cellular validation and model organism confirmation; multi-family replication","pmids":["41887223"],"is_preprint":false}],"current_model":"OLA1 is a universally conserved P-loop ATPase (preferring ATP over GTP) that functions as a multifunctional regulatory hub: it inhibits translation initiation by binding eIF2 and interfering with ternary complex formation; stabilizes HSP70 by blocking CHIP-mediated ubiquitination; regulates centrosome number and maturation through a complex with BRCA1/BARD1/γ-tubulin, where BARD1's BRCT domain allosterically activates OLA1 ATPase and Aurora A/NEK2-mediated ubiquitination controls centrosomal OLA1 abundance; suppresses antioxidant responses via nontranscriptional ROS modulation; modulates GSK3β activity to influence Snail-dependent EMT and HIF1α signaling; undergoes phosphorylation-dependent nuclear translocation (Ser232/Tyr236/Thr325) to drive mitochondrial gene expression; is N-terminally methylated by NTMT1; and interacts with STING to suppress innate immune signaling, with its disruption causing a human neurodevelopmental disorder with joint hypermobility."},"narrative":{"mechanistic_narrative":"OLA1 is a universally conserved P-loop ATPase of the Obg/YchF subfamily that preferentially binds and hydrolyzes ATP over GTP, a specificity rationalized by its AMPPCP-bound crystal structure [PMID:17430889], and which acts as a multifunctional regulatory hub coupling nucleotide hydrolysis to translational, proteostatic, centrosomal, and redox control. In translation, OLA1 binds eIF2 and interferes with ternary complex (eIF2-GTP-tRNAi) formation, inhibiting protein synthesis and promoting the integrated stress response; loss of OLA1 yields a hypoactive ISR and enhanced translation of targets such as p21, and Ola1-knockout mice show growth retardation and cell-cycle defects [PMID:26283179, PMID:27481995]. Through its chaperone-like activity, OLA1 binds the HSP70 C-terminal variable domain to block CHIP-mediated ubiquitination and degradation of HSP70, an interaction extended to protection of the Hsp70–SOD2 complex from CHIP-driven turnover [PMID:23412384, PMID:31476900]. At centrosomes and spindle poles, OLA1 forms a complex with the N-terminus of BRCA1, with γ-tubulin, and with the BARD1 C-terminus; its loss causes centrosome amplification, and BARD1's BRCT domain binds the OLA1 TGS domain to allosterically increase ATPase turnover, while cancer-associated mutations (OLA1 E168Q; BARD1 V695L) disrupt binding and abolish centrosome-number rescue [PMID:24289923, PMID:29858377, PMID:35134491]. Centrosomal OLA1 abundance is set by Aurora A-mediated polyubiquitination enhanced by NEK2 phosphorylation at T124, controlling pericentriolar material recruitment during centrosome maturation [PMID:37481721]. OLA1 also negatively regulates the cellular antioxidant response and ROS through nontranscriptional mechanisms [PMID:19706404], binds and inhibits GSK3β via Ser9 phosphorylation to stabilize Snail and promote TGF-β-induced EMT and to modulate HIF1α/CA9 signaling [PMID:26863455, PMID:35440019], and undergoes ERK1/2-driven phosphorylation at Ser232/Tyr236 and Thr325 that triggers nuclear translocation and an ATPase-to-GTPase switch driving expression of nuclear-encoded mitochondrial bioenergetic genes [PMID:36481055]. OLA1 interacts with STING to suppress its activation, and competing STING engagement liberates Keap1 to destabilize Nrf2, linking OLA1 to innate immune and ferroptosis regulation [PMID:38132845, PMID:41352507]. Bi-allelic loss-of-function variants in OLA1 cause a human neurodevelopmental disorder with joint hypermobility, with patient cells and model organisms showing impaired migration, adhesion, and cytoskeletal/microtubule dynamics via FAK [PMID:41887223].","teleology":[{"year":2007,"claim":"Established the fundamental biochemical identity of OLA1, resolving whether this Obg-family protein is a GTPase or an ATPase.","evidence":"Nucleotide-binding/hydrolysis assays and an AMPPCP-bound X-ray structure of human OLA1","pmids":["17430889"],"confidence":"High","gaps":["Did not establish a cellular substrate or pathway for the ATPase activity","Functional consequence of nucleotide hydrolysis in vivo unaddressed"]},{"year":2009,"claim":"Defined OLA1 as a nontranscriptional negative regulator of the antioxidant response, distinguishing it from canonical transcription-driven redox control.","evidence":"Bidirectional RNAi/overexpression with ROS and viability assays plus cycloheximide block and qRT-PCR in human cells; migration/invasion link in breast cancer cells","pmids":["19706404","19882753"],"confidence":"High","gaps":["Molecular target mediating ROS suppression not identified","Connection between ROS control and motility correlative"]},{"year":2013,"claim":"Placed OLA1 in two distinct protein complexes — a centrosomal BRCA1/BARD1/γ-tubulin module and an HSP70-stabilizing chaperone role — defining its first direct physical partners.","evidence":"Mass spectrometry, reciprocal Co-IP, direct binding, immunofluorescence, RNAi, and mutant rescue (E168Q); ubiquitination and thermotolerance assays for HSP70/CHIP","pmids":["24289923","23412384"],"confidence":"High","gaps":["How ATPase activity couples to centrosome regulation unresolved","Whether HSP70 and centrosome roles are mechanistically related unknown"]},{"year":2015,"claim":"Demonstrated that OLA1 inhibits translation initiation and drives the integrated stress response by binding eIF2 and blocking ternary complex formation, with tumor-suppressive consequences in vivo.","evidence":"Co-IP, GTPase and ternary complex formation assays, polysome profiling, RNAi, and xenograft models","pmids":["26283179"],"confidence":"High","gaps":["Stoichiometry and regulation of OLA1–eIF2 binding undefined","Relationship to the centrosomal pool of OLA1 unclear"]},{"year":2016,"claim":"Showed OLA1 is genetically required for mammalian development and connects translational control to cell-cycle progression via eIF2-dependent p21 translation, and separately identified GSK3β as a partner driving EMT.","evidence":"Ola1 knockout mice, primary MEFs, double-knockout epistasis, and eIF2α pharmacology; Co-IP and EMT assays for GSK3β/Snail","pmids":["27481995","26863455"],"confidence":"High","gaps":["How OLA1 mechanistically restrains eIF2α-dependent p21 translation unresolved","Direct effect of OLA1 on GSK3β Ser9 phosphorylation mechanism unclear"]},{"year":2019,"claim":"Extended the OLA1–HSP70 chaperone axis to disease-relevant SOD2 protection and identified OLA1 as a substrate of N-terminal methyltransferase NTMT1.","evidence":"OLA1 KO mice and CHIP-Hsp70-SOD2 Co-IP/ubiquitination in a PPHN model; activity-based substrate profiling and NTMT1 CRISPR KO validation","pmids":["31476900","31857877"],"confidence":"Medium","gaps":["Functional consequence of N-terminal methylation on OLA1 activity not established","Whether SOD2 protection is direct or via HSP70 stabilization not fully separated"]},{"year":2020,"claim":"Identified meiotic spindle functions for OLA1 and revealed that viral and host factors converge on the OLA1–GSK3β axis to control autophagy.","evidence":"Immunofluorescence, nocodazole, siRNA microinjection in mouse oocytes; Co-IP and autophagy/proliferation assays for HIV p17 disruption of OLA1–GSK3β","pmids":["31915569","32790080"],"confidence":"Medium","gaps":["Mechanism linking OLA1 to SAC timing not defined","Single-lab ortholog/viral models without reciprocal validation"]},{"year":2021,"claim":"Demonstrated that a lncRNA (ZFAS1) can allosterically enhance OLA1 ATPase activity by exposing its ATP-binding motif, coupling OLA1 enzymatic output to cancer metabolism.","evidence":"RNA pulldown, RIP, ATP hydrolysis and glycolysis assays with ATP-site mutagenesis in colorectal cancer cells","pmids":["34743750"],"confidence":"Medium","gaps":["Whether ZFAS1 regulation operates in non-cancer contexts unknown","Single lab"]},{"year":2022,"claim":"Resolved BARD1 as an allosteric ATPase-activating protein for OLA1 and connected OLA1 to the GSK3β–HIF1α/CA9 hypoxic axis.","evidence":"Enzyme kinetics, 1.88 Å crystal structure of BARD1 BRCT V695L, and mutagenesis; CRISPR KO with mRNA-seq and xenografts for HIF1α/CA9","pmids":["35134491","35440019"],"confidence":"High","gaps":["How allosteric activation feeds into centrosome regulation mechanistically incomplete","No direct OLA1–HIF1α interaction shown"]},{"year":2023,"claim":"Defined the ubiquitin-mediated control of centrosomal OLA1 abundance and a phosphorylation-driven nuclear translocation that switches OLA1 between ATPase and GTPase activities to control mitochondrial gene expression.","evidence":"In vitro ubiquitination, T124 mutagenesis, proteasome inhibition, and centrosome imaging (Aurora A/NEK2); phospho-site mutagenesis, fractionation, activity assays, ERK/PP1A manipulation and KO mice (Ser232/Tyr236/Thr325)","pmids":["37481721","36481055"],"confidence":"High","gaps":["Kinase(s) directly phosphorylating Ser232/Tyr236 not fully defined","How a single protein partitions between nuclear, mitochondrial and centrosomal roles unresolved"]},{"year":2024,"claim":"Established OLA1 as a suppressor of STING-mediated innate immune signaling and a target of HIV-1 p17 antagonism.","evidence":"Co-IP of OLA1–STING and p17–OLA1, cGAMP stimulation, STING translocation/phosphorylation, and comparative HIV-1 vs HIV-2/SIV p17 with activity assays","pmids":["38132845"],"confidence":"Medium","gaps":["Whether ATPase/GTPase activity is required for STING suppression unclear","Single lab, no reciprocal in vivo validation"]},{"year":2025,"claim":"Connected OLA1 to redox and cell-death regulation through a STING–OLA1–Keap1–Nrf2 axis and via MAPK/mitochondrial ROS control in a yeast ortholog.","evidence":"Co-IP of STING–OLA1 and OLA1–Keap1 with Nrf2 stability/ferroptosis readouts in a murine POF model; Co-IP and mtROS assays for S. pombe Ola1–Pmk1/Pek1","pmids":["41352507","40543417"],"confidence":"Medium","gaps":["Direct biochemical basis of OLA1–Keap1 competition with STING undefined","Conservation of yeast MAPK mechanism to mammals untested"]},{"year":2026,"claim":"Established OLA1 as a Mendelian disease gene, linking its loss of function to a neurodevelopmental disorder with joint hypermobility via cytoskeletal/FAK-dependent defects.","evidence":"Exome sequencing across 14 individuals/9 families, proband fibroblast and iPSC-neuron assays, and C. elegans ola-1 knockout with transcriptomics","pmids":["41887223"],"confidence":"Medium","gaps":["Which of OLA1's many molecular activities underlies the disorder unresolved","Mechanistic link between OLA1 and FAK levels not biochemically established"]},{"year":null,"claim":"How a single conserved ATPase coordinates its many partner-specific roles — translation, chaperoning, centrosome regulation, redox, and mitochondrial gene expression — into a unified mechanism remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No integrated model explaining context-dependent partner selection","Whether nucleotide hydrolysis is required across all functional contexts unknown","Spatial/temporal partitioning between subcellular pools undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0,14,15,18]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,6]},{"term_id":"GO:0044183","term_label":"protein folding chaperone","supporting_discovery_ids":[4,11]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[4,6,7,15,19]},{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[6,18]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[3,17]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[18]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[18]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[18]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[3,12]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[4,6,8]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[1,6]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[3,8,17]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[19]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,16,21]}],"complexes":["BRCA1/BARD1/γ-tubulin centrosomal complex"],"partners":["BRCA1","BARD1","TUBG1","HSPA1A","EIF2","GSK3B","STING1","KEAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NTK5","full_name":"Obg-like ATPase 1","aliases":["DNA damage-regulated overexpressed in cancer 45","DOC45","GTP-binding protein 9"],"length_aa":396,"mass_kda":44.7,"function":"Hydrolyzes ATP, and can also hydrolyze GTP with lower efficiency. Has lower affinity for GTP","subcellular_location":"Cytoplasm; Nucleus; Nucleus, nucleolus","url":"https://www.uniprot.org/uniprotkb/Q9NTK5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/OLA1","classification":"Not Classified","n_dependent_lines":40,"n_total_lines":1208,"dependency_fraction":0.033112582781456956},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"SAR1B","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/OLA1","total_profiled":1310},"omim":[{"mim_id":"611175","title":"OBG-LIKE ATPase 1; OLA1","url":"https://www.omim.org/entry/611175"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Cytosol","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/OLA1"},"hgnc":{"alias_symbol":["PTD004"],"prev_symbol":["GTPBP9"]},"alphafold":{"accession":"Q9NTK5","domains":[{"cath_id":"3.40.50.300","chopping":"23-301","consensus_level":"medium","plddt":89.0069,"start":23,"end":301},{"cath_id":"3.10.20.30","chopping":"305-386","consensus_level":"medium","plddt":96.3243,"start":305,"end":386}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NTK5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NTK5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NTK5-F1-predicted_aligned_error_v6.png","plddt_mean":88.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=OLA1","jax_strain_url":"https://www.jax.org/strain/search?query=OLA1"},"sequence":{"accession":"Q9NTK5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NTK5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NTK5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NTK5"}},"corpus_meta":[{"pmid":"34743750","id":"PMC_34743750","title":"N6-methyladenosine reader IMP2 stabilizes the ZFAS1/OLA1 axis and activates the Warburg effect: implication in colorectal cancer.","date":"2021","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34743750","citation_count":97,"is_preprint":false},{"pmid":"17430889","id":"PMC_17430889","title":"Human OLA1 defines an ATPase subfamily in the Obg family of GTP-binding proteins.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17430889","citation_count":92,"is_preprint":false},{"pmid":"19706404","id":"PMC_19706404","title":"OLA1, an Obg-like ATPase, suppresses antioxidant response via nontranscriptional mechanisms.","date":"2009","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/19706404","citation_count":63,"is_preprint":false},{"pmid":"24289923","id":"PMC_24289923","title":"The BRCA1/BARD1-interacting protein OLA1 functions in centrosome regulation.","date":"2013","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/24289923","citation_count":61,"is_preprint":false},{"pmid":"23412384","id":"PMC_23412384","title":"OLA1 protects cells in heat shock by stabilizing HSP70.","date":"2013","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/23412384","citation_count":57,"is_preprint":false},{"pmid":"26283179","id":"PMC_26283179","title":"OLA1 regulates protein synthesis and integrated stress response by inhibiting eIF2 ternary complex formation.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26283179","citation_count":39,"is_preprint":false},{"pmid":"19882753","id":"PMC_19882753","title":"Knockdown of OLA1, a regulator of oxidative stress response, inhibits motility and invasion of breast cancer cells.","date":"2009","source":"Journal of Zhejiang University. Science. B","url":"https://pubmed.ncbi.nlm.nih.gov/19882753","citation_count":35,"is_preprint":false},{"pmid":"26863455","id":"PMC_26863455","title":"OLA1 contributes to epithelial-mesenchymal transition in lung cancer by modulating the GSK3β/snail/E-cadherin signaling.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26863455","citation_count":32,"is_preprint":false},{"pmid":"27481995","id":"PMC_27481995","title":"OLA1, a Translational Regulator of p21, Maintains Optimal Cell Proliferation Necessary for Developmental Progression.","date":"2016","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/27481995","citation_count":30,"is_preprint":false},{"pmid":"31476900","id":"PMC_31476900","title":"Decreased OLA1 (Obg-Like ATPase-1) Expression Drives Ubiquitin-Proteasome Pathways to Downregulate Mitochondrial SOD2 (Superoxide Dismutase) in Persistent Pulmonary Hypertension of the Newborn.","date":"2019","source":"Hypertension (Dallas, Tex. : 1979)","url":"https://pubmed.ncbi.nlm.nih.gov/31476900","citation_count":29,"is_preprint":false},{"pmid":"29858377","id":"PMC_29858377","title":"BRCA1-Interacting Protein OLA1 Requires Interaction with BARD1 to Regulate Centrosome Number.","date":"2018","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/29858377","citation_count":23,"is_preprint":false},{"pmid":"24486488","id":"PMC_24486488","title":"Regulation of cell-matrix adhesion by OLA1, the Obg-like ATPase 1.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/24486488","citation_count":18,"is_preprint":false},{"pmid":"35440019","id":"PMC_35440019","title":"OLA1 promotes colorectal cancer tumorigenesis by activation of HIF1α/CA9 axis.","date":"2022","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35440019","citation_count":12,"is_preprint":false},{"pmid":"31857877","id":"PMC_31857877","title":"In vivo methylation of OLA1 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chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37856434","citation_count":6,"is_preprint":false},{"pmid":"24411079","id":"PMC_24411079","title":"OLA1 in centrosome biology alongside the BRCA1/BARD1 complex: looking beyond centrosomes.","date":"2014","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/24411079","citation_count":6,"is_preprint":false},{"pmid":"26984302","id":"PMC_26984302","title":"Metal mixture (As-Cd-Pb)-induced cell transformation is modulated by OLA1.","date":"2016","source":"Mutagenesis","url":"https://pubmed.ncbi.nlm.nih.gov/26984302","citation_count":5,"is_preprint":false},{"pmid":"32790080","id":"PMC_32790080","title":"HIV p17 enhances T cell proliferation by suppressing autophagy through the p17-OLA1-GSK3β axis under nutrient starvation.","date":"2020","source":"Journal of medical virology","url":"https://pubmed.ncbi.nlm.nih.gov/32790080","citation_count":5,"is_preprint":false},{"pmid":"31915569","id":"PMC_31915569","title":"OLA1 is responsible for normal spindle assembly and SAC activation in mouse oocytes.","date":"2020","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/31915569","citation_count":4,"is_preprint":false},{"pmid":"38132845","id":"PMC_38132845","title":"HIV-1 p17 matrix protein enhances type I interferon responses through the p17-OLA1-STING axis.","date":"2024","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/38132845","citation_count":3,"is_preprint":false},{"pmid":"36232807","id":"PMC_36232807","title":"Association of Common Variants in OLA1 Gene with Preclinical Atherosclerosis.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36232807","citation_count":3,"is_preprint":false},{"pmid":"38889130","id":"PMC_38889130","title":"Identification and development of Tetra-ARMS PCR-based screening test for a genetic variant of OLA1 (Tyr254Cys) in the human failing heart.","date":"2024","source":"PloS 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Molecular basis of disease","url":"https://pubmed.ncbi.nlm.nih.gov/38537683","citation_count":0,"is_preprint":false},{"pmid":"41887223","id":"PMC_41887223","title":"Bi-allelic variants in OLA1 cause a neurodevelopmental disorder with joint hypermobility.","date":"2026","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/41887223","citation_count":0,"is_preprint":false},{"pmid":"38602870","id":"PMC_38602870","title":"Protocol to detect OLA1 polyubiquitination by Aurora A in vivo and in vitro.","date":"2024","source":"STAR protocols","url":"https://pubmed.ncbi.nlm.nih.gov/38602870","citation_count":0,"is_preprint":false},{"pmid":"41352507","id":"PMC_41352507","title":"The STING-OLA1-Keap1-Nrf2 axis regulates ferroptosis in premature ovarian failure.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/41352507","citation_count":0,"is_preprint":false},{"pmid":"41246329","id":"PMC_41246329","title":"Anti-OLA1 autoantibody is a potential early diagnostic marker for hepatocellular carcinoma.","date":"2025","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/41246329","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":19191,"output_tokens":6468,"usd":0.077297,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15487,"output_tokens":5288,"usd":0.104817,"stage2_stop_reason":"end_turn"},"total_usd":0.182114,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"Human OLA1 (hOLA1) binds and hydrolyzes ATP more efficiently than GTP, establishing it as an ATPase rather than a GTPase within the Obg/YchF subfamily. X-ray crystal structure of hOLA1 bound to the non-hydrolyzable ATP analogue AMPPCP explained the altered nucleotide specificity compared to other Obg-family GTPases.\",\n      \"method\": \"Biochemical nucleotide-binding and hydrolysis assays; X-ray crystallography with AMPPCP-bound structure\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus in vitro enzymatic characterization, replicated across multiple YchF orthologs in the same study\",\n      \"pmids\": [\"17430889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"OLA1 functions as a negative regulator of the cellular antioxidant response through nontranscriptional mechanisms. Knockdown increased resistance to oxidizing agents (tBH, diamide) and reduced intracellular ROS and glutathione depletion without altering antioxidant gene mRNA levels or requiring de novo protein synthesis.\",\n      \"method\": \"RNAi knockdown; OLA1 overexpression; cell viability and ROS assays; cycloheximide block experiments; qRT-PCR of antioxidant genes\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional manipulation (KD and OE), multiple orthogonal readouts, mechanistic dissection ruling out transcriptional mechanism\",\n      \"pmids\": [\"19706404\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Knockdown of OLA1 inhibits breast cancer cell migration and invasion through modulation of intracellular ROS levels, linking OLA1's regulation of ROS to cytoskeletal motility.\",\n      \"method\": \"siRNA knockdown; wound-healing and transwell invasion assays; ROS measurement; N-acetylcysteine treatment\",\n      \"journal\": \"Journal of Zhejiang University. Science. B\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, multiple functional assays but no direct molecular partner identified for motility phenotype\",\n      \"pmids\": [\"19882753\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"OLA1 directly binds to the amino-terminal region of BRCA1 and to γ-tubulin; it interacts with the carboxy-terminal region of BARD1. OLA1 localizes to centrosomes in interphase and to the spindle pole in mitosis. OLA1 knockdown causes centrosome amplification and activation of microtubule aster formation. A cancer-derived mutation E168Q abrogates BRCA1 binding and fails to rescue centrosome amplification.\",\n      \"method\": \"Mass spectrometry identification; co-immunoprecipitation; direct binding assays; immunofluorescence localization; RNAi knockdown; rescue with mutant constructs\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, direct binding, localization with functional consequence, mutagenesis rescue experiments\",\n      \"pmids\": [\"24289923\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"OLA1 stabilizes HSP70 by binding to its carboxyl-terminus variable domain, thereby blocking recruitment of the E3 ubiquitin ligase CHIP and preventing CHIP-mediated ubiquitination and degradation of HSP70. OLA1 downregulation reduces steady-state HSP70 levels and impairs heat-shock-induced HSP70 induction, increasing cellular sensitivity to heat shock.\",\n      \"method\": \"RNAi knockdown; targeted gene disruption; OLA1 overexpression; protein-protein interaction (co-IP/pulldown); ubiquitination assay; thermal resistance assay\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional manipulation, direct interaction assay, ubiquitination mechanistic assay, functional thermotolerance readout in single lab\",\n      \"pmids\": [\"23412384\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"OLA1 negatively regulates cell-matrix adhesion and spreading. OLA1-deficient cells have elevated FAK protein levels and decreased Ser3 phosphorylation of cofilin, while OLA1-overexpressing cells show opposite changes, indicating OLA1 modulates adhesion at least partly through FAK expression and cofilin phosphorylation.\",\n      \"method\": \"RNAi knockdown; OLA1 overexpression; cell adhesion/spreading assays; western blot for FAK and p-cofilin\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — bidirectional manipulation with two molecular readouts, single lab, no direct interaction between OLA1 and FAK/cofilin demonstrated\",\n      \"pmids\": [\"24486488\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"OLA1 inhibits protein synthesis and promotes the integrated stress response (ISR) by binding eIF2, hydrolyzing GTP in that context, and interfering with ternary complex (eIF2-GTP-tRNAi) formation. OLA1 depletion causes hypoactive ISR, reduces CHOP induction, and promotes tumor growth and metastasis in vivo.\",\n      \"method\": \"Co-immunoprecipitation (OLA1-eIF2 interaction); GTPase activity assay; ternary complex formation assay; polysome profiling; RNAi knockdown; xenograft tumor models\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding to eIF2, enzymatic activity assay, TC formation assay, in vivo xenograft validation; multiple orthogonal methods in single lab\",\n      \"pmids\": [\"26283179\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"OLA1 is a GSK3β-interacting protein and inhibits GSK3β activity by mediating its Ser9 phosphorylation, thereby suppressing GSK3β-mediated degradation of Snail, which promotes E-cadherin downregulation and contributes to TGF-β-induced epithelial-mesenchymal transition (EMT).\",\n      \"method\": \"Co-immunoprecipitation (OLA1-GSK3β); western blot for GSK3β Ser9 phosphorylation, Snail, and E-cadherin; RNAi knockdown; EMT assays with TGF-β treatment\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP plus phosphorylation readout and EMT functional assays, single lab\",\n      \"pmids\": [\"26863455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"OLA1 is required for normal mammalian development. Ola1-knockout mouse embryos have growth retardation and developmental delay. Primary Ola1-/- MEFs show impaired proliferation due to defective cell-cycle progression, with reduced cyclins D1/E1, attenuated Rb phosphorylation, and elevated p21 protein. p21 accumulation is due to enhanced mRNA translation that is reversed by eIF2α dephosphorylation inhibitor, placing OLA1 upstream of eIF2-mediated p21 translational control.\",\n      \"method\": \"Ola1 knockout mice; primary MEF culture; BrdU/cell-cycle analysis; western blot; polysome/translation assays; eIF2α pharmacological manipulation; p21-/-/Ola1-/- double-knockout rescue\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic knockout, double-knockout epistasis, pharmacological rescue, multiple molecular and cellular readouts\",\n      \"pmids\": [\"27481995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"OLA1 requires its interaction with BARD1 to properly regulate centrosome number. Three OLA1 missense mutants that fail to bind BARD1 are deficient in centrosome number regulation. Phosphomimetic mutations at specific OLA1 residues restore BARD1 binding and rescue centrosome amplification. BARD1 mutant V695L (cancer-associated) fails to bind OLA1 and cannot rescue BARD1 knockdown-induced centrosome amplification.\",\n      \"method\": \"Co-immunoprecipitation; site-directed mutagenesis of OLA1 (phosphorylation, acetylation, ATP-binding residues); overexpression of mutant constructs; RNAi knockdown/rescue; centrosome counting by immunofluorescence\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic mutagenesis, binding assays, functional rescue experiments, bidirectional manipulation of BARD1\",\n      \"pmids\": [\"29858377\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"OLA1 is N-terminally methylated in vivo by the N-terminal methyltransferase NTMT1, as demonstrated by activity-based substrate profiling and validation in NTMT1 knockout cells.\",\n      \"method\": \"Activity-based substrate profiling with Hey-SAM analogue; CRISPR-Cas9 NTMT1 knockout HEK293FT cells; mass spectrometry validation\",\n      \"journal\": \"Chemical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chemical biology profiling plus genetic KO validation, single lab\",\n      \"pmids\": [\"31857877\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Decreased OLA1 expression in PPHN enhances CHIP affinity for the Hsp70-SOD2 complex, facilitating SOD2 ubiquitination and proteasomal degradation. OLA1 acts as a molecular chaperone whose stress-induced activity prevents CHIP-mediated SOD2 degradation; ola1-/- mice recapitulate PPHN phenotypes including SOD2 downregulation and pulmonary vascular remodeling.\",\n      \"method\": \"Patient/fetal lamb tissue analysis; OLA1 KO mice; co-immunoprecipitation of CHIP-Hsp70-SOD2; ubiquitination assays; right ventricular pressure measurement\",\n      \"journal\": \"Hypertension (Dallas, Tex. : 1979)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO in vivo, interaction assays, disease model; single lab\",\n      \"pmids\": [\"31476900\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"OLA1 localizes to meiotic spindles in mouse oocytes (co-localizing with spindle structures after GVBD, confirmed by nocodazole treatment). OLA1 knockdown results in abnormal/multipolar spindle assembly, premature anaphase onset due to precocious spindle assembly checkpoint (SAC) inactivation, and impaired germinal vesicle breakdown.\",\n      \"method\": \"Immunofluorescence/confocal microscopy; nocodazole treatment; siRNA microinjection; chromosome spreading; polar body extrusion assays\",\n      \"journal\": \"PeerJ\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — localization with functional consequence, RNAi knockdown with multiple readouts, single lab\",\n      \"pmids\": [\"31915569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"HIV p17 interacts with OLA1 and disrupts the OLA1-GSK3β complex, leading to GSK3β hyperactivation, suppression of autophagy, and enhanced proliferation of HIV-infected T cells under glucose starvation conditions.\",\n      \"method\": \"Co-immunoprecipitation (p17-OLA1, OLA1-GSK3β); autophagy flux assays; T cell proliferation assays under glucose starvation\",\n      \"journal\": \"Journal of medical virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP interaction plus functional autophagy and proliferation readouts, single lab\",\n      \"pmids\": [\"32790080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ZFAS1 lncRNA recognizes the OBG-type functional domain of OLA1, facilitating exposure of its ATP-binding site (NVGKST, residues 32–37), enhancing OLA1 protein ATPase activity, and accelerating ATP hydrolysis and the Warburg effect in colorectal cancer cells.\",\n      \"method\": \"RNA pulldown; RIP assay; ATP hydrolysis assay; ECAR/glycolysis assay; mutagenesis of OLA1 ATP-binding site\",\n      \"journal\": \"Journal of hematology & oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — RNA-protein interaction assay plus enzymatic activity measurement, single lab\",\n      \"pmids\": [\"34743750\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"BARD1 acts as an ATPase activating protein (AAP) for OLA1. The BARD1 BRCT domain binds the OLA1 TGS domain via a conserved BUDR motif and allosterically increases OLA1 ATPase turnover number (kcat). Cancer-associated BARD1 mutation V695L reduces OLA1 binding and activation, as revealed by a 1.88 Å crystal structure of the V695L BRCT mutant.\",\n      \"method\": \"Enzyme kinetics assays; X-ray crystallography (BRCT V695L mutant at 1.88 Å); biophysical/biochemical binding assays; mutagenesis\",\n      \"journal\": \"Biochimica et biophysica acta. General subjects\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure plus enzyme kinetics plus mutagenesis, multiple orthogonal methods in single study\",\n      \"pmids\": [\"35134491\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"OLA1 knockout in colorectal cancer cells activates GSK3β and downregulates HIF1α and its target CA9 at the mRNA/protein level, linking OLA1 to the HIF1α/CA9 hypoxic signaling axis through GSK3β.\",\n      \"method\": \"CRISPR-Cas9 OLA1 knockout; mRNA sequencing; western blot for GSK3β, HIF1α, CA9; xenograft tumor models\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — genetic KO with pathway readouts, single lab, no direct OLA1-HIF1α interaction demonstrated\",\n      \"pmids\": [\"35440019\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Aurora A binds OLA1 and polyubiquitinates it, targeting OLA1 for proteasomal degradation. NEK2 phosphorylates OLA1 at T124, which increases OLA1 binding to Aurora A and enhances Aurora A-mediated polyubiquitination. The kinase activity of Aurora A suppresses its own E3 ligase activity toward OLA1. Reduction of centrosomal OLA1 by this mechanism promotes pericentriolar material protein recruitment in G2 phase, required for centrosome maturation.\",\n      \"method\": \"Co-immunoprecipitation; in vitro ubiquitination assay; mutagenesis (T124 phosphorylation site); proteasome inhibitor experiments; immunofluorescence of centrosomal OLA1; overexpression rescue\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro ubiquitination reconstitution, mutagenesis, in vivo degradation assays, centrosome functional readout; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"37481721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"OLA1 phosphorylation at Ser232/Tyr236 triggers its translocation from cytoplasm/mitochondria into the nucleus. Subsequent phosphorylation at Thr325 switches its biochemical activity from ATPase to GTPase and promotes expression of nuclear-encoded mitochondrial bioenergetic genes. ERK1/2 drives this process and is restrained by PP1A. OLA1 T325A (phosphoresistant) mutant blocks nuclear translocation and compromises mitochondrial gene expression. OLA1 knockout mice have fewer mitochondria, lower ATP, and higher lactate.\",\n      \"method\": \"Phospho-site mutagenesis; subcellular fractionation; immunofluorescence; ATPase/GTPase activity assays with phosphomimetic mutants; ERK inhibition; PP1A manipulation; OLA1 KO mice; metabolite measurements\",\n      \"journal\": \"American journal of respiratory cell and molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — mutagenesis of phosphorylation sites, enzymatic activity switch demonstrated, KO mice with metabolic phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"36481055\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"HIV-1 p17 (but not HIV-2 or SIV p17) binds OLA1 and inhibits OLA1's interaction with STING, thereby blocking OLA1-mediated suppression of STING translocation and phosphorylation. OLA1 normally interacts with STING and inhibits STING activation upon cGAMP stimulation. HIV-1 p17 also specifically promotes OLA1 ATPase and GTPase activities.\",\n      \"method\": \"Co-immunoprecipitation (OLA1-STING, p17-OLA1); cGAMP stimulation assays; STING translocation/phosphorylation assays; comparative HIV-1 vs HIV-2/SIV p17 experiments; ATPase/GTPase activity assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP plus functional STING signaling assays, comparative species analysis; single lab\",\n      \"pmids\": [\"38132845\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In fission yeast (S. pombe), Ola1 physically interacts with MAPK/Pmk1 and its upstream kinase Pek1 (MAPKK), inhibiting MAPK/Pmk1 signaling to prevent excessive mitochondrial ROS accumulation. Absence of Ola1 increases mtROS, promotes nuclear localization of Hsf1, and upregulates Ssa1 (Hsp70 homolog).\",\n      \"method\": \"Co-immunoprecipitation (Ola1-Pmk1, Ola1-Pek1); mitochondrial ROS measurements; ola1 deletion; Hsf1 localization assay; western blot\",\n      \"journal\": \"Microbiological research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP with functional mtROS readout in a yeast ortholog model; single lab\",\n      \"pmids\": [\"40543417\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"OLA1 interacts with Keap1 and, when STING is activated, enhanced STING-OLA1 interaction disrupts the OLA1-Keap1 complex, liberating Keap1 to promote Nrf2 degradation and ferroptosis in granulosa cells. This STING-OLA1-Keap1-Nrf2 axis is mechanistically linked to premature ovarian failure.\",\n      \"method\": \"Co-immunoprecipitation (STING-OLA1, OLA1-Keap1); siRNA knockdown; Nrf2 protein stability assays; ferroptosis markers; murine POF model; molecular docking (Icariin-STING)\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP interactions with functional Nrf2/ferroptosis readouts in cellular and in vivo models; single lab\",\n      \"pmids\": [\"41352507\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Vitexin binds OLA1 (identified by tissue thermal proteome profiling and molecular docking) and the OLA1-vitexin complex interacts with Keap1, disrupting the Keap1-Nrf2 interaction and activating Nrf2. siRNA knockdown of OLA1 in Caco-2 cells confirmed OLA1's role in mediating Nrf2 protein expression and anti-inflammatory effects.\",\n      \"method\": \"Tissue thermal proteome profiling; molecular docking; siRNA knockdown; Nrf2 protein level assays; inflammatory cytokine measurements\",\n      \"journal\": \"Journal of agricultural and food chemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3–4 / Weak — computational docking plus single siRNA KD experiment; no direct biochemical reconstitution of OLA1-Keap1 interaction\",\n      \"pmids\": [\"37856434\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Bi-allelic loss-of-function variants in OLA1 cause a human neurodevelopmental disorder with joint hypermobility. Patient-derived fibroblasts recapitulate impaired migration and proliferation. Neurons derived from proband fibroblasts show impaired adhesion and cytoskeletal control. In C. elegans, ola-1 deficiency reduces neurite numbers and suppresses microtubule dynamics and axon regrowth, placing OLA1 in a pathway regulating cytoskeletal dynamics through FAK levels.\",\n      \"method\": \"Sanger/exome sequencing in 14 individuals from 9 families; proband-derived fibroblast migration/proliferation assays; iPSC-derived neuron adhesion/cytoskeletal assays; C. elegans ola-1 knockout; transcriptomics\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — patient genetic evidence plus proband cellular validation and model organism confirmation; multi-family replication\",\n      \"pmids\": [\"41887223\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"OLA1 is a universally conserved P-loop ATPase (preferring ATP over GTP) that functions as a multifunctional regulatory hub: it inhibits translation initiation by binding eIF2 and interfering with ternary complex formation; stabilizes HSP70 by blocking CHIP-mediated ubiquitination; regulates centrosome number and maturation through a complex with BRCA1/BARD1/γ-tubulin, where BARD1's BRCT domain allosterically activates OLA1 ATPase and Aurora A/NEK2-mediated ubiquitination controls centrosomal OLA1 abundance; suppresses antioxidant responses via nontranscriptional ROS modulation; modulates GSK3β activity to influence Snail-dependent EMT and HIF1α signaling; undergoes phosphorylation-dependent nuclear translocation (Ser232/Tyr236/Thr325) to drive mitochondrial gene expression; is N-terminally methylated by NTMT1; and interacts with STING to suppress innate immune signaling, with its disruption causing a human neurodevelopmental disorder with joint hypermobility.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"OLA1 is a universally conserved P-loop ATPase of the Obg/YchF subfamily that preferentially binds and hydrolyzes ATP over GTP, a specificity rationalized by its AMPPCP-bound crystal structure [#0], and which acts as a multifunctional regulatory hub coupling nucleotide hydrolysis to translational, proteostatic, centrosomal, and redox control. In translation, OLA1 binds eIF2 and interferes with ternary complex (eIF2-GTP-tRNAi) formation, inhibiting protein synthesis and promoting the integrated stress response; loss of OLA1 yields a hypoactive ISR and enhanced translation of targets such as p21, and Ola1-knockout mice show growth retardation and cell-cycle defects [#6, #8]. Through its chaperone-like activity, OLA1 binds the HSP70 C-terminal variable domain to block CHIP-mediated ubiquitination and degradation of HSP70, an interaction extended to protection of the Hsp70–SOD2 complex from CHIP-driven turnover [#4, #11]. At centrosomes and spindle poles, OLA1 forms a complex with the N-terminus of BRCA1, with \\u03b3-tubulin, and with the BARD1 C-terminus; its loss causes centrosome amplification, and BARD1's BRCT domain binds the OLA1 TGS domain to allosterically increase ATPase turnover, while cancer-associated mutations (OLA1 E168Q; BARD1 V695L) disrupt binding and abolish centrosome-number rescue [#3, #9, #15]. Centrosomal OLA1 abundance is set by Aurora A-mediated polyubiquitination enhanced by NEK2 phosphorylation at T124, controlling pericentriolar material recruitment during centrosome maturation [#17]. OLA1 also negatively regulates the cellular antioxidant response and ROS through nontranscriptional mechanisms [#1], binds and inhibits GSK3\\u03b2 via Ser9 phosphorylation to stabilize Snail and promote TGF-\\u03b2-induced EMT and to modulate HIF1\\u03b1/CA9 signaling [#7, #16], and undergoes ERK1/2-driven phosphorylation at Ser232/Tyr236 and Thr325 that triggers nuclear translocation and an ATPase-to-GTPase switch driving expression of nuclear-encoded mitochondrial bioenergetic genes [#18]. OLA1 interacts with STING to suppress its activation, and competing STING engagement liberates Keap1 to destabilize Nrf2, linking OLA1 to innate immune and ferroptosis regulation [#19, #21]. Bi-allelic loss-of-function variants in OLA1 cause a human neurodevelopmental disorder with joint hypermobility, with patient cells and model organisms showing impaired migration, adhesion, and cytoskeletal/microtubule dynamics via FAK [#23].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established the fundamental biochemical identity of OLA1, resolving whether this Obg-family protein is a GTPase or an ATPase.\",\n      \"evidence\": \"Nucleotide-binding/hydrolysis assays and an AMPPCP-bound X-ray structure of human OLA1\",\n      \"pmids\": [\"17430889\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish a cellular substrate or pathway for the ATPase activity\", \"Functional consequence of nucleotide hydrolysis in vivo unaddressed\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined OLA1 as a nontranscriptional negative regulator of the antioxidant response, distinguishing it from canonical transcription-driven redox control.\",\n      \"evidence\": \"Bidirectional RNAi/overexpression with ROS and viability assays plus cycloheximide block and qRT-PCR in human cells; migration/invasion link in breast cancer cells\",\n      \"pmids\": [\"19706404\", \"19882753\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular target mediating ROS suppression not identified\", \"Connection between ROS control and motility correlative\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Placed OLA1 in two distinct protein complexes — a centrosomal BRCA1/BARD1/\\u03b3-tubulin module and an HSP70-stabilizing chaperone role — defining its first direct physical partners.\",\n      \"evidence\": \"Mass spectrometry, reciprocal Co-IP, direct binding, immunofluorescence, RNAi, and mutant rescue (E168Q); ubiquitination and thermotolerance assays for HSP70/CHIP\",\n      \"pmids\": [\"24289923\", \"23412384\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How ATPase activity couples to centrosome regulation unresolved\", \"Whether HSP70 and centrosome roles are mechanistically related unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrated that OLA1 inhibits translation initiation and drives the integrated stress response by binding eIF2 and blocking ternary complex formation, with tumor-suppressive consequences in vivo.\",\n      \"evidence\": \"Co-IP, GTPase and ternary complex formation assays, polysome profiling, RNAi, and xenograft models\",\n      \"pmids\": [\"26283179\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and regulation of OLA1–eIF2 binding undefined\", \"Relationship to the centrosomal pool of OLA1 unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Showed OLA1 is genetically required for mammalian development and connects translational control to cell-cycle progression via eIF2-dependent p21 translation, and separately identified GSK3\\u03b2 as a partner driving EMT.\",\n      \"evidence\": \"Ola1 knockout mice, primary MEFs, double-knockout epistasis, and eIF2\\u03b1 pharmacology; Co-IP and EMT assays for GSK3\\u03b2/Snail\",\n      \"pmids\": [\"27481995\", \"26863455\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How OLA1 mechanistically restrains eIF2\\u03b1-dependent p21 translation unresolved\", \"Direct effect of OLA1 on GSK3\\u03b2 Ser9 phosphorylation mechanism unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended the OLA1–HSP70 chaperone axis to disease-relevant SOD2 protection and identified OLA1 as a substrate of N-terminal methyltransferase NTMT1.\",\n      \"evidence\": \"OLA1 KO mice and CHIP-Hsp70-SOD2 Co-IP/ubiquitination in a PPHN model; activity-based substrate profiling and NTMT1 CRISPR KO validation\",\n      \"pmids\": [\"31476900\", \"31857877\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of N-terminal methylation on OLA1 activity not established\", \"Whether SOD2 protection is direct or via HSP70 stabilization not fully separated\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified meiotic spindle functions for OLA1 and revealed that viral and host factors converge on the OLA1–GSK3\\u03b2 axis to control autophagy.\",\n      \"evidence\": \"Immunofluorescence, nocodazole, siRNA microinjection in mouse oocytes; Co-IP and autophagy/proliferation assays for HIV p17 disruption of OLA1–GSK3\\u03b2\",\n      \"pmids\": [\"31915569\", \"32790080\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking OLA1 to SAC timing not defined\", \"Single-lab ortholog/viral models without reciprocal validation\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstrated that a lncRNA (ZFAS1) can allosterically enhance OLA1 ATPase activity by exposing its ATP-binding motif, coupling OLA1 enzymatic output to cancer metabolism.\",\n      \"evidence\": \"RNA pulldown, RIP, ATP hydrolysis and glycolysis assays with ATP-site mutagenesis in colorectal cancer cells\",\n      \"pmids\": [\"34743750\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ZFAS1 regulation operates in non-cancer contexts unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Resolved BARD1 as an allosteric ATPase-activating protein for OLA1 and connected OLA1 to the GSK3\\u03b2–HIF1\\u03b1/CA9 hypoxic axis.\",\n      \"evidence\": \"Enzyme kinetics, 1.88 \\u00c5 crystal structure of BARD1 BRCT V695L, and mutagenesis; CRISPR KO with mRNA-seq and xenografts for HIF1\\u03b1/CA9\",\n      \"pmids\": [\"35134491\", \"35440019\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How allosteric activation feeds into centrosome regulation mechanistically incomplete\", \"No direct OLA1–HIF1\\u03b1 interaction shown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined the ubiquitin-mediated control of centrosomal OLA1 abundance and a phosphorylation-driven nuclear translocation that switches OLA1 between ATPase and GTPase activities to control mitochondrial gene expression.\",\n      \"evidence\": \"In vitro ubiquitination, T124 mutagenesis, proteasome inhibition, and centrosome imaging (Aurora A/NEK2); phospho-site mutagenesis, fractionation, activity assays, ERK/PP1A manipulation and KO mice (Ser232/Tyr236/Thr325)\",\n      \"pmids\": [\"37481721\", \"36481055\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinase(s) directly phosphorylating Ser232/Tyr236 not fully defined\", \"How a single protein partitions between nuclear, mitochondrial and centrosomal roles unresolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established OLA1 as a suppressor of STING-mediated innate immune signaling and a target of HIV-1 p17 antagonism.\",\n      \"evidence\": \"Co-IP of OLA1–STING and p17–OLA1, cGAMP stimulation, STING translocation/phosphorylation, and comparative HIV-1 vs HIV-2/SIV p17 with activity assays\",\n      \"pmids\": [\"38132845\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ATPase/GTPase activity is required for STING suppression unclear\", \"Single lab, no reciprocal in vivo validation\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected OLA1 to redox and cell-death regulation through a STING–OLA1–Keap1–Nrf2 axis and via MAPK/mitochondrial ROS control in a yeast ortholog.\",\n      \"evidence\": \"Co-IP of STING–OLA1 and OLA1–Keap1 with Nrf2 stability/ferroptosis readouts in a murine POF model; Co-IP and mtROS assays for S. pombe Ola1–Pmk1/Pek1\",\n      \"pmids\": [\"41352507\", \"40543417\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical basis of OLA1–Keap1 competition with STING undefined\", \"Conservation of yeast MAPK mechanism to mammals untested\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Established OLA1 as a Mendelian disease gene, linking its loss of function to a neurodevelopmental disorder with joint hypermobility via cytoskeletal/FAK-dependent defects.\",\n      \"evidence\": \"Exome sequencing across 14 individuals/9 families, proband fibroblast and iPSC-neuron assays, and C. elegans ola-1 knockout with transcriptomics\",\n      \"pmids\": [\"41887223\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Which of OLA1's many molecular activities underlies the disorder unresolved\", \"Mechanistic link between OLA1 and FAK levels not biochemically established\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single conserved ATPase coordinates its many partner-specific roles — translation, chaperoning, centrosome regulation, redox, and mitochondrial gene expression — into a unified mechanism remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No integrated model explaining context-dependent partner selection\", \"Whether nucleotide hydrolysis is required across all functional contexts unknown\", \"Spatial/temporal partitioning between subcellular pools undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0, 14, 15, 18]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 6]},\n      {\"term_id\": \"GO:0044183\", \"supporting_discovery_ids\": [4, 11]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [4, 6, 7, 15, 19]},\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [6, 18]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3, 17]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [18]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [18]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [18]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [3, 12]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [4, 6, 8]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [1, 6]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [3, 8, 17]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [19]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 16, 21]}\n    ],\n    \"complexes\": [\"BRCA1/BARD1/\\u03b3-tubulin centrosomal complex\"],\n    \"partners\": [\"BRCA1\", \"BARD1\", \"TUBG1\", \"HSPA1A\", \"EIF2\", \"GSK3B\", \"STING1\", \"KEAP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}