{"gene":"ANKZF1","run_date":"2026-04-28T17:12:37","timeline":{"discoveries":[{"year":2018,"finding":"ANKZF1 (human ortholog of yeast Vms1) is a peptidyl-tRNA hydrolase that induces specific cleavage in the tRNA acceptor arm of ubiquitinated nascent chain-tRNA/60S ribosomal complexes (60S RNCs), releasing proteasome-degradable ubiquitinated nascent chains linked to four 3'-terminal tRNA nucleotides. This activity requires NEMF- and Listerin-dependent ubiquitination of nascent chains, which accommodates the NC-tRNA in the P site and renders 60S RNCs resistant to Ptrh1 but susceptible to ANKZF1.","method":"In vitro reconstitution with purified components, peptidyl-tRNA hydrolase assay, tRNA cleavage mapping","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with defined components, cleavage site mapped, activity dependent on upstream ubiquitination state","pmids":["30244831"],"is_preprint":false},{"year":2018,"finding":"Yeast Vms1 (founding member of the Vms1-like release factor 1 clade, of which ANKZF1 is the human ortholog) functions as a peptidyl-tRNA hydrolase on 60S ribosomal subunits carrying stalled nascent chains; its activity depends on a conserved catalytic glutamine residue analogous to eukaryotic release factor 1 (eRF1). Vms1 is a Cdc48 adaptor that cleaves the tRNA from ubiquitylated nascent chains prior to proteasomal degradation.","method":"In vitro peptidyl-tRNA hydrolase assay, active-site mutagenesis (catalytic Gln), evolutionary sequence analysis, Co-IP","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — in vitro enzymatic assay with mutagenesis, replicated across multiple labs","pmids":["29632312"],"is_preprint":false},{"year":2019,"finding":"Cryo-EM structures of yeast Vms1 (ANKZF1 ortholog) bound to 60S ribosomal subunits in pre- and post-cleavage states reveal that Vms1 binds via its VLRF1, zinc finger, and ankyrin domains. The VLRF1 domain projects its catalytic GSQ motif toward the CCA end of the P-site tRNA, and residue Y285 dislodges tRNA A73 to enable nucleolytic cleavage. The ABCF-type ATPase Arb1 occupies the E-site in the pre-cleavage state, stabilizing the delocalized A73 and stimulating Vms1-dependent tRNA cleavage.","method":"Cryo-EM structure determination, functional mutagenesis, in vitro cleavage assay","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 — high-resolution cryo-EM with functional validation and mutagenesis","pmids":["31189955"],"is_preprint":false},{"year":2017,"finding":"Yeast Vms1 (ANKZF1 ortholog) binds to 60S ribosomes at the mitochondrial surface and antagonizes Rqc2-mediated CAT-tailing of stalled nascent chains targeted to mitochondria, thereby facilitating mitochondrial import and directing aberrant polypeptides to intra-mitochondrial quality control. In the absence of Vms1, CAT-tailed polypeptides aggregate after import and sequester mitochondrial chaperones (e.g., Hsp78) and translation machinery.","method":"Genetic deletion, co-immunoprecipitation, ribosome fractionation, in vivo aggregation assay, mitochondrial import assay","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods, highly cited foundational paper","pmids":["29107329"],"is_preprint":false},{"year":2017,"finding":"Mitochondrial targeting of yeast Vms1 (ANKZF1 ortholog) is mediated by a conserved mitochondrial targeting domain (MTD) that is held in an autoinhibited state through intramolecular binding to the Vms1 leucine-rich sequence (LRS). The oxidized sterol ergosterol peroxide binds the MTD competitively with the LRS to relieve autoinhibition and trigger Vms1 translocation to stressed mitochondria.","method":"2.7 Å crystal structure, biochemical binding assay, sterol binding competition assay, live-cell fluorescence localization","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — crystal structure combined with biochemical and cell biological validation","pmids":["29149595"],"is_preprint":false},{"year":2013,"finding":"Yeast Vms1 (ANKZF1 ortholog) mitochondrial targeting is negatively regulated by an intramolecular interaction between its N-terminal segment and the mitochondrial targeting domain (MTD). Vms1 is preferentially recruited to mitochondria under oxidative stress, as shown by laser-induced mitochondrial ROS generation.","method":"Truncation mutants, live-cell fluorescence imaging, laser-induced oxidative stress, biochemical binding assay","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment with functional consequence, single lab","pmids":["23468520"],"is_preprint":false},{"year":2012,"finding":"Yeast Vms1 (ANKZF1 ortholog) and its binding partner Cdc48 (but not Ufd1 or Ufd2) are required for degradation of Cdc13, a telomere-capping protein. Both autophagy and the proteasome contribute to Cdc13 turnover, and accumulation of Cdc13 in vms1Δ cells causes toxicity.","method":"Genetic deletion, protein degradation assay, epistasis with autophagy mutants","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis with defined substrate and phenotypic readout, single lab","pmids":["22718752"],"is_preprint":false},{"year":2014,"finding":"The yeast Cdc48-Vms1 complex (ANKZF1 ortholog) is required for maintenance of 26S proteasome architecture. Loss of Vms1 leads to accumulation of unassembled 20S core particles and select 19S cap subunits, reduced 26S proteasome levels, accumulation of ubiquitinated proteins, and decreased viability in stationary phase. Vms1's support of proteasome assembly requires its interaction with Cdc48.","method":"Genetic deletion, native gel electrophoresis of proteasome complexes, ubiquitinated protein accumulation assay, viability assay","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 — clean KO with multiple cellular phenotype readouts, single lab","pmids":["24351022"],"is_preprint":false},{"year":2024,"finding":"ANKZF1 knockdown in glioblastoma cells causes accumulation of CAT-tailed polypeptides in mitochondria, activating the mitochondrial unfolded protein response (UPRmt). Excess CAT-tails sequester mitochondrial chaperones HSP60 and mtHSP70, protease LONP1, and respiratory chain subunits ND1, Cytb, mtCO2, and ATP6, resulting in oxidative phosphorylation dysfunction, membrane potential impairment, and activation of the mitochondrial apoptotic pathway.","method":"siRNA knockdown, co-immunoprecipitation, mitochondrial fractionation, membrane potential assay, apoptosis assay, xenograft model","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function with defined molecular and cellular phenotypes in human cells, single lab","pmids":["38670305"],"is_preprint":false},{"year":2025,"finding":"Human ANKZF1 participates in PINK1-Parkin-mediated mitophagy. ANKZF1 is recruited to damaged mitochondria together with Parkin during proteotoxic stress or membrane depolarization. ANKZF1 physically interacts with Parkin and LC3, and LIR motif 4 (residues 333–336) is required for ANKZF1–LC3 interaction. ANKZF1 knockout cells are defective in clearing stress-damaged mitochondria. ANKZF1 functions as a mitophagy adaptor bridging polyubiquitinated outer mitochondrial membrane proteins (via its UBA domain) and autophagosome receptor LC3 (via its LIR motif).","method":"ANKZF1 knockout, live-cell fluorescence imaging, co-immunoprecipitation, LIR mutagenesis, mitophagy flux assay","journal":"Cell death discovery","confidence":"Medium","confidence_rationale":"Tier 2 — KO with functional readout, interaction validated by Co-IP, domain mutagenesis, single lab","pmids":["40730577"],"is_preprint":false},{"year":2026,"finding":"The N-terminal 73 residues of human ANKZF1 negatively regulate its mitochondrial targeting by suppressing an internal matrix-targeting sequence-like sequence (iMTS-L) at residues 231–240. Deletion of the N-terminal segment causes structural rearrangement (shown by MD simulation) that exposes the 231–240 iMTS-L. Residues 231–324 constitute an independent mitochondrial signal that can target GFP to mitochondria when fused to its N-terminus.","method":"Truncation mutants, live-cell fluorescence localization, GFP-fusion targeting assay, molecular dynamics simulation","journal":"The FEBS journal","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiments with functional mutants and structural modeling, single lab","pmids":["41920018"],"is_preprint":false},{"year":2024,"finding":"ANKZF1 interacts with YWHAE (14-3-3ε) to competitively inhibit cytoplasmic retention of YAP1, thereby promoting YAP1 nuclear import and transcriptional activation of pro-lymphangiogenic factors in clear-cell renal cell carcinoma. NAT10-mediated ac4C modification of ANKZF1 mRNA upregulates ANKZF1 expression.","method":"Co-immunoprecipitation, immunofluorescence, site-directed mutagenesis, RNA immunoprecipitation, mass spectrometry","journal":"Cancer communications","confidence":"Medium","confidence_rationale":"Tier 3 — Co-IP with mechanistic follow-up in cancer cell context, single lab","pmids":["38407929"],"is_preprint":false}],"current_model":"ANKZF1 (human ortholog of yeast Vms1) is a peptidyl-tRNA hydrolase that cleaves the tRNA acceptor arm of ubiquitinated nascent chain–tRNA complexes on stalled 60S ribosomal subunits during ribosome-associated quality control (RQC), releasing proteasome-degradable ubiquitinated nascent chains; its mitochondrial targeting is autoinhibited by the N-terminal domain and relieved by stress signals, whereupon ANKZF1 antagonizes CAT-tailing to protect mitochondria from toxic polypeptide aggregation, and also participates in PINK1-Parkin-mediated mitophagy as an adaptor bridging ubiquitinated outer mitochondrial membrane proteins to LC3."},"narrative":{"teleology":[{"year":2012,"claim":"Establishing that Vms1/ANKZF1 cooperates with Cdc48/p97 in protein degradation pathways revealed its initial functional link to ubiquitin-proteasome quality control beyond a generic stress factor.","evidence":"Genetic deletion of VMS1 in yeast with Cdc13 degradation assay and epistasis analysis with autophagy mutants","pmids":["22718752"],"confidence":"Medium","gaps":["Substrate specificity beyond Cdc13 unknown","Direct enzymatic activity not yet identified","Mechanism of Vms1-Cdc48 cooperation unresolved"]},{"year":2013,"claim":"Demonstrating that the N-terminal domain autoinhibits mitochondrial targeting of Vms1, with oxidative stress relieving this block, established the regulated translocation mechanism that controls ANKZF1 function at mitochondria.","evidence":"Truncation mutants and live-cell imaging with laser-induced mitochondrial ROS in yeast","pmids":["23468520"],"confidence":"Medium","gaps":["Identity of the stress-induced signal relieving autoinhibition not determined","Structural basis of intramolecular autoinhibition unknown"]},{"year":2014,"claim":"Showing that the Cdc48-Vms1 complex is required for 26S proteasome assembly revealed an unexpected role in maintaining the proteolytic machinery itself, beyond being a simple substrate adaptor.","evidence":"VMS1 deletion in yeast with native gel analysis of proteasome complexes and ubiquitin accumulation assays","pmids":["24351022"],"confidence":"Medium","gaps":["Direct versus indirect role in proteasome biogenesis unclear","Not confirmed in mammalian cells"]},{"year":2017,"claim":"Two concurrent studies resolved the core mitochondrial function of Vms1/ANKZF1: it binds 60S ribosomes at the mitochondrial surface to antagonize Rqc2-mediated CAT-tailing, preventing toxic aggregation of stalled nascent chains inside mitochondria, and its mitochondrial translocation is controlled by an autoinhibitory N-terminal–MTD interaction relieved by the oxidized sterol ergosterol peroxide.","evidence":"Crystal structure of MTD–sterol complex, competitive binding assays, genetic deletion with mitochondrial import and aggregation assays, ribosome fractionation in yeast","pmids":["29107329","29149595"],"confidence":"High","gaps":["Whether identical sterol-based activation applies to human ANKZF1 is unresolved","How Vms1 selects mitochondrial versus cytoplasmic stalled ribosomes unknown"]},{"year":2018,"claim":"Identification of ANKZF1/Vms1 as a peptidyl-tRNA hydrolase that specifically cleaves the tRNA acceptor arm on ubiquitinated 60S RNCs defined its catalytic activity, showing it depends on a conserved glutamine (analogous to eRF1) and requires upstream NEMF/Listerin-dependent ubiquitination to render the substrate susceptible.","evidence":"In vitro reconstitution with purified components, active-site mutagenesis, tRNA cleavage mapping in both yeast and human systems","pmids":["30244831","29632312"],"confidence":"High","gaps":["Kinetic parameters of the hydrolase activity uncharacterized","Whether additional cofactors modulate activity in vivo unknown"]},{"year":2019,"claim":"Cryo-EM structures of Vms1 on the 60S subunit in pre- and post-cleavage states revealed how the VLRF1 domain positions its GSQ catalytic motif at the CCA end of P-site tRNA, how Y285 dislodges tRNA A73 to enable cleavage, and how the E-site ATPase Arb1 stimulates this reaction.","evidence":"Cryo-EM structure determination with functional mutagenesis and in vitro cleavage assays in yeast","pmids":["31189955"],"confidence":"High","gaps":["No equivalent high-resolution structure of human ANKZF1 on 60S","Role of Arb1 ortholog in human RQC not tested"]},{"year":2024,"claim":"Loss-of-function studies in human glioblastoma cells demonstrated that ANKZF1 depletion causes CAT-tail accumulation in mitochondria, sequestering chaperones (HSP60, mtHSP70) and respiratory chain subunits, activating the UPRmt and triggering mitochondrial apoptosis—providing the first direct evidence of ANKZF1's anti-CAT-tail function in human cells.","evidence":"siRNA knockdown with mitochondrial fractionation, co-IP, membrane potential assay, and xenograft model in human glioblastoma cells","pmids":["38670305"],"confidence":"Medium","gaps":["Findings from a single cancer cell type; generalizability to non-malignant cells untested","Whether ANKZF1 acts catalytically or structurally to suppress CAT-tailing in human cells not resolved"]},{"year":2025,"claim":"Discovery that ANKZF1 acts as a mitophagy adaptor during PINK1-Parkin-mediated mitophagy, bridging ubiquitinated outer mitochondrial membrane proteins to LC3 through its UBA domain and LIR motif (residues 333–336), expanded its function beyond RQC to organelle clearance.","evidence":"ANKZF1 knockout, co-IP of ANKZF1 with Parkin and LC3, LIR mutagenesis, and mitophagy flux assay in human cells","pmids":["40730577"],"confidence":"Medium","gaps":["Single-lab finding; independent confirmation needed","Relative contribution of ANKZF1 versus other mitophagy receptors (OPTN, NDP52) not compared","Whether RQC and mitophagy adaptor functions are coordinated or independent is unclear"]},{"year":2026,"claim":"Mapping the human ANKZF1 N-terminal autoinhibition to residues 1–73 suppressing an internal matrix-targeting sequence at residues 231–240 provided the first structural rationale for regulated mitochondrial import of the human protein.","evidence":"Truncation mutants, GFP-fusion targeting assays, and molecular dynamics simulation in human cells","pmids":["41920018"],"confidence":"Medium","gaps":["MD simulation-based structural model lacks experimental structural validation","Endogenous signal that relieves N-terminal autoinhibition in human cells not identified"]},{"year":null,"claim":"Key open questions remain: the identity of the physiological signal relieving ANKZF1 autoinhibition in human cells, a high-resolution structure of human ANKZF1 on the 60S subunit, whether the RQC peptidyl-tRNA hydrolase and mitophagy adaptor functions are coordinated, and the in vivo substrate spectrum of ANKZF1 beyond stalled nascent chains.","evidence":"","pmids":[],"confidence":"High","gaps":["No high-resolution structure of human ANKZF1 on 60S ribosome","Physiological activating signal in human cells unidentified","Functional coordination between RQC and mitophagy roles untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[9]}],"localization":[{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[3,4,5,8,10]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[3,5]}],"pathway":[{"term_id":"GO:0005840","term_label":"ribosome","supporting_discovery_ids":[0,1,2,3]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,2,3,8]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[9]}],"complexes":["60S RQC complex"],"partners":["NEMF","LTN1","VCP","PRKN","MAP1LC3B","YWHAE"],"other_free_text":[]},"mechanistic_narrative":"ANKZF1 is a peptidyl-tRNA hydrolase central to ribosome-associated quality control (RQC) that cleaves the tRNA acceptor arm of ubiquitinated nascent chain–tRNA complexes on stalled 60S ribosomal subunits, releasing proteasome-degradable nascent chains [PMID:30244831, PMID:29632312]. Its catalytic mechanism depends on a conserved GSQ motif within the VLRF1 domain, which is positioned toward the CCA end of the P-site tRNA, with an ABCF-type ATPase (Arb1) stimulating cleavage in the pre-cleavage state [PMID:31189955]. At mitochondria, ANKZF1 antagonizes CAT-tailing of stalled nascent chains to prevent toxic polypeptide aggregation and sequestration of mitochondrial chaperones and respiratory chain subunits; its mitochondrial targeting is autoinhibited by the N-terminal domain, which masks an internal matrix-targeting sequence relieved by stress signals such as oxidized sterols [PMID:29107329, PMID:29149595, PMID:38670305, PMID:41920018]. ANKZF1 also functions as a mitophagy adaptor during PINK1-Parkin-mediated mitophagy, bridging polyubiquitinated outer mitochondrial membrane proteins (via its UBA domain) to LC3 on autophagosomes (via a LIR motif) [PMID:40730577]."},"prefetch_data":{"uniprot":{"accession":"Q9H8Y5","full_name":"tRNA endonuclease ANKZF1","aliases":["Ankyrin repeat and zinc finger domain-containing protein 1","Zinc finger protein 744"],"length_aa":726,"mass_kda":80.9,"function":"Endonuclease that cleaves polypeptidyl-tRNAs downstream of the ribosome-associated quality control (RQC) pathway to release incompletely synthesized polypeptides for degradation (PubMed:29632312, PubMed:30244831, PubMed:31011209). The RQC pathway disassembles aberrantly stalled translation complexes to recycle or degrade the constituent parts (PubMed:29632312, PubMed:30244831, PubMed:31011209). ANKZF1 acts downstream disassembly of stalled ribosomes and specifically cleaves off the terminal 3'-CCA nucleotides universal to all tRNAs from polypeptidyl-tRNAs, releasing (1) ubiquitinated polypeptides from 60S ribosomal subunit for degradation and (2) cleaved tRNAs (PubMed:31011209). ANKZF1-cleaved tRNAs are then repaired and recycled by ELAC1 and TRNT1 (PubMed:31011209, PubMed:32075755). Also plays a role in the cellular response to hydrogen peroxide and in the maintenance of mitochondrial integrity under conditions of cellular stress (PubMed:28302725)","subcellular_location":"Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q9H8Y5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ANKZF1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000163516","cell_line_id":"CID001398","localizations":[{"compartment":"cytoplasmic","grade":3}],"interactors":[{"gene":"PSMD12","stoichiometry":0.2},{"gene":"PSMD11","stoichiometry":0.2},{"gene":"PSMD3","stoichiometry":0.2},{"gene":"PSMC4","stoichiometry":0.2},{"gene":"PSMB2","stoichiometry":0.2},{"gene":"PSMD7","stoichiometry":0.2},{"gene":"PSMA5","stoichiometry":0.2},{"gene":"PSMC6","stoichiometry":0.2},{"gene":"APRT","stoichiometry":0.2},{"gene":"PSMC3","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID001398","total_profiled":1310},"omim":[{"mim_id":"621047","title":"PEPTIDYL-tRNA HYDROLASE 1; PTRH1","url":"https://www.omim.org/entry/621047"},{"mim_id":"617541","title":"ANKYRIN REPEAT- AND ZINC FINGER DOMAIN-CONTAINING 1; ANKZF1","url":"https://www.omim.org/entry/617541"},{"mim_id":"612326","title":"TRANSCRIPTION FACTOR 25; TCF25","url":"https://www.omim.org/entry/612326"},{"mim_id":"601023","title":"VALOSIN-CONTAINING PROTEIN; VCP","url":"https://www.omim.org/entry/601023"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Nuclear bodies","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/ANKZF1"},"hgnc":{"alias_symbol":["FLJ10415","ZNF744","Vms1"],"prev_symbol":[]},"alphafold":{"accession":"Q9H8Y5","domains":[{"cath_id":"-","chopping":"79-112","consensus_level":"high","plddt":86.7718,"start":79,"end":112},{"cath_id":"3.30.420.60","chopping":"165-352","consensus_level":"medium","plddt":84.4491,"start":165,"end":352},{"cath_id":"1.25.40.20","chopping":"495-607","consensus_level":"high","plddt":83.0433,"start":495,"end":607},{"cath_id":"-","chopping":"612-723","consensus_level":"medium","plddt":83.3362,"start":612,"end":723}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H8Y5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H8Y5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H8Y5-F1-predicted_aligned_error_v6.png","plddt_mean":68.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ANKZF1","jax_strain_url":"https://www.jax.org/strain/search?query=ANKZF1"},"sequence":{"accession":"Q9H8Y5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9H8Y5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9H8Y5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H8Y5"}},"corpus_meta":[{"pmid":"29107329","id":"PMC_29107329","title":"Cytosolic Protein Vms1 Links Ribosome Quality Control to Mitochondrial and Cellular Homeostasis.","date":"2017","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/29107329","citation_count":163,"is_preprint":false},{"pmid":"29632312","id":"PMC_29632312","title":"Vms1 and ANKZF1 peptidyl-tRNA hydrolases release nascent chains from stalled ribosomes.","date":"2018","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/29632312","citation_count":134,"is_preprint":false},{"pmid":"30244831","id":"PMC_30244831","title":"Release of Ubiquitinated and Non-ubiquitinated Nascent Chains from Stalled Mammalian Ribosomal Complexes by ANKZF1 and Ptrh1.","date":"2018","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/30244831","citation_count":96,"is_preprint":false},{"pmid":"31189955","id":"PMC_31189955","title":"Structure and function of Vms1 and Arb1 in RQC and mitochondrial proteome homeostasis.","date":"2019","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/31189955","citation_count":78,"is_preprint":false},{"pmid":"29149595","id":"PMC_29149595","title":"Sterol Oxidation Mediates Stress-Responsive Vms1 Translocation to Mitochondria.","date":"2017","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/29149595","citation_count":35,"is_preprint":false},{"pmid":"38407929","id":"PMC_38407929","title":"NAT10-mediated ac4C-modified ANKZF1 promotes tumor progression and lymphangiogenesis in clear-cell renal cell carcinoma by attenuating YWHAE-driven cytoplasmic retention of YAP1.","date":"2024","source":"Cancer communications (London, England)","url":"https://pubmed.ncbi.nlm.nih.gov/38407929","citation_count":29,"is_preprint":false},{"pmid":"23468520","id":"PMC_23468520","title":"Intramolecular interactions control Vms1 translocation to damaged mitochondria.","date":"2013","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/23468520","citation_count":26,"is_preprint":false},{"pmid":"36344988","id":"PMC_36344988","title":"The expression changes of transcription factors including ANKZF1, LEF1, CASZ1, and ATOH1 as a predictor of survival rate in colorectal cancer: a large-scale analysis.","date":"2022","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/36344988","citation_count":15,"is_preprint":false},{"pmid":"22718752","id":"PMC_22718752","title":"The Cdc48 protein and its cofactor Vms1 are involved in Cdc13 protein degradation.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22718752","citation_count":13,"is_preprint":false},{"pmid":"38670305","id":"PMC_38670305","title":"ANKZF1 knockdown inhibits glioblastoma progression by promoting intramitochondrial protein aggregation through mitoRQC.","date":"2024","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/38670305","citation_count":9,"is_preprint":false},{"pmid":"24351022","id":"PMC_24351022","title":"The Cdc48-Vms1 complex maintains 26S proteasome architecture.","date":"2014","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/24351022","citation_count":7,"is_preprint":false},{"pmid":"38666541","id":"PMC_38666541","title":"Overexpression of ZNF169 promotes the growth and proliferation of colorectal cancer cells via the upregulation of ANKZF1.","date":"2024","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/38666541","citation_count":5,"is_preprint":false},{"pmid":"29203248","id":"PMC_29203248","title":"Vms1: A Cytosolic CAT-Tailing Antagonist to Protect Mitochondria.","date":"2017","source":"Trends in cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29203248","citation_count":4,"is_preprint":false},{"pmid":"29112848","id":"PMC_29112848","title":"Vms1 Relieves a Mitochondrial Import Chokehold.","date":"2017","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/29112848","citation_count":3,"is_preprint":false},{"pmid":"40730577","id":"PMC_40730577","title":"ANKZF1 helps to eliminate stress-damaged mitochondria by LC3-mediated mitophagy.","date":"2025","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/40730577","citation_count":1,"is_preprint":false},{"pmid":"41920018","id":"PMC_41920018","title":"The N-terminal segment of the human ANKZF1 negatively regulates its internal mitochondrial targeting signal to prevent localization to mitochondria.","date":"2026","source":"The FEBS journal","url":"https://pubmed.ncbi.nlm.nih.gov/41920018","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.04.04.647180","title":"The N-terminal Segment of the Human ANKZF1 negatively regulates its internal mitochondrial targeting signal to prevent its mitochondrial localization","date":"2025-04-05","source":"bioRxiv","url":"https://doi.org/10.1101/2025.04.04.647180","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.09.04.673968","title":"Jlp2 is an RQC complex-independent release factor acting on aberrant peptidyl-tRNA, protecting cells against translation elongation stress","date":"2025-09-06","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.04.673968","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10054,"output_tokens":3294,"usd":0.039786},"stage2":{"model":"claude-opus-4-6","input_tokens":6665,"output_tokens":2770,"usd":0.153863},"total_usd":0.193649,"stage1_batch_id":"msgbatch_01TqNM6p6FsoMBT2jzQTwKBr","stage2_batch_id":"msgbatch_01Ya473eco8sTHbvRymQaU8o","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2018,\n      \"finding\": \"ANKZF1 (human ortholog of yeast Vms1) is a peptidyl-tRNA hydrolase that induces specific cleavage in the tRNA acceptor arm of ubiquitinated nascent chain-tRNA/60S ribosomal complexes (60S RNCs), releasing proteasome-degradable ubiquitinated nascent chains linked to four 3'-terminal tRNA nucleotides. This activity requires NEMF- and Listerin-dependent ubiquitination of nascent chains, which accommodates the NC-tRNA in the P site and renders 60S RNCs resistant to Ptrh1 but susceptible to ANKZF1.\",\n      \"method\": \"In vitro reconstitution with purified components, peptidyl-tRNA hydrolase assay, tRNA cleavage mapping\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with defined components, cleavage site mapped, activity dependent on upstream ubiquitination state\",\n      \"pmids\": [\"30244831\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Yeast Vms1 (founding member of the Vms1-like release factor 1 clade, of which ANKZF1 is the human ortholog) functions as a peptidyl-tRNA hydrolase on 60S ribosomal subunits carrying stalled nascent chains; its activity depends on a conserved catalytic glutamine residue analogous to eukaryotic release factor 1 (eRF1). Vms1 is a Cdc48 adaptor that cleaves the tRNA from ubiquitylated nascent chains prior to proteasomal degradation.\",\n      \"method\": \"In vitro peptidyl-tRNA hydrolase assay, active-site mutagenesis (catalytic Gln), evolutionary sequence analysis, Co-IP\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzymatic assay with mutagenesis, replicated across multiple labs\",\n      \"pmids\": [\"29632312\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Cryo-EM structures of yeast Vms1 (ANKZF1 ortholog) bound to 60S ribosomal subunits in pre- and post-cleavage states reveal that Vms1 binds via its VLRF1, zinc finger, and ankyrin domains. The VLRF1 domain projects its catalytic GSQ motif toward the CCA end of the P-site tRNA, and residue Y285 dislodges tRNA A73 to enable nucleolytic cleavage. The ABCF-type ATPase Arb1 occupies the E-site in the pre-cleavage state, stabilizing the delocalized A73 and stimulating Vms1-dependent tRNA cleavage.\",\n      \"method\": \"Cryo-EM structure determination, functional mutagenesis, in vitro cleavage assay\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — high-resolution cryo-EM with functional validation and mutagenesis\",\n      \"pmids\": [\"31189955\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Yeast Vms1 (ANKZF1 ortholog) binds to 60S ribosomes at the mitochondrial surface and antagonizes Rqc2-mediated CAT-tailing of stalled nascent chains targeted to mitochondria, thereby facilitating mitochondrial import and directing aberrant polypeptides to intra-mitochondrial quality control. In the absence of Vms1, CAT-tailed polypeptides aggregate after import and sequester mitochondrial chaperones (e.g., Hsp78) and translation machinery.\",\n      \"method\": \"Genetic deletion, co-immunoprecipitation, ribosome fractionation, in vivo aggregation assay, mitochondrial import assay\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods, highly cited foundational paper\",\n      \"pmids\": [\"29107329\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mitochondrial targeting of yeast Vms1 (ANKZF1 ortholog) is mediated by a conserved mitochondrial targeting domain (MTD) that is held in an autoinhibited state through intramolecular binding to the Vms1 leucine-rich sequence (LRS). The oxidized sterol ergosterol peroxide binds the MTD competitively with the LRS to relieve autoinhibition and trigger Vms1 translocation to stressed mitochondria.\",\n      \"method\": \"2.7 Å crystal structure, biochemical binding assay, sterol binding competition assay, live-cell fluorescence localization\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure combined with biochemical and cell biological validation\",\n      \"pmids\": [\"29149595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Yeast Vms1 (ANKZF1 ortholog) mitochondrial targeting is negatively regulated by an intramolecular interaction between its N-terminal segment and the mitochondrial targeting domain (MTD). Vms1 is preferentially recruited to mitochondria under oxidative stress, as shown by laser-induced mitochondrial ROS generation.\",\n      \"method\": \"Truncation mutants, live-cell fluorescence imaging, laser-induced oxidative stress, biochemical binding assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment with functional consequence, single lab\",\n      \"pmids\": [\"23468520\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Yeast Vms1 (ANKZF1 ortholog) and its binding partner Cdc48 (but not Ufd1 or Ufd2) are required for degradation of Cdc13, a telomere-capping protein. Both autophagy and the proteasome contribute to Cdc13 turnover, and accumulation of Cdc13 in vms1Δ cells causes toxicity.\",\n      \"method\": \"Genetic deletion, protein degradation assay, epistasis with autophagy mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with defined substrate and phenotypic readout, single lab\",\n      \"pmids\": [\"22718752\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"The yeast Cdc48-Vms1 complex (ANKZF1 ortholog) is required for maintenance of 26S proteasome architecture. Loss of Vms1 leads to accumulation of unassembled 20S core particles and select 19S cap subunits, reduced 26S proteasome levels, accumulation of ubiquitinated proteins, and decreased viability in stationary phase. Vms1's support of proteasome assembly requires its interaction with Cdc48.\",\n      \"method\": \"Genetic deletion, native gel electrophoresis of proteasome complexes, ubiquitinated protein accumulation assay, viability assay\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean KO with multiple cellular phenotype readouts, single lab\",\n      \"pmids\": [\"24351022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ANKZF1 knockdown in glioblastoma cells causes accumulation of CAT-tailed polypeptides in mitochondria, activating the mitochondrial unfolded protein response (UPRmt). Excess CAT-tails sequester mitochondrial chaperones HSP60 and mtHSP70, protease LONP1, and respiratory chain subunits ND1, Cytb, mtCO2, and ATP6, resulting in oxidative phosphorylation dysfunction, membrane potential impairment, and activation of the mitochondrial apoptotic pathway.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation, mitochondrial fractionation, membrane potential assay, apoptosis assay, xenograft model\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined molecular and cellular phenotypes in human cells, single lab\",\n      \"pmids\": [\"38670305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Human ANKZF1 participates in PINK1-Parkin-mediated mitophagy. ANKZF1 is recruited to damaged mitochondria together with Parkin during proteotoxic stress or membrane depolarization. ANKZF1 physically interacts with Parkin and LC3, and LIR motif 4 (residues 333–336) is required for ANKZF1–LC3 interaction. ANKZF1 knockout cells are defective in clearing stress-damaged mitochondria. ANKZF1 functions as a mitophagy adaptor bridging polyubiquitinated outer mitochondrial membrane proteins (via its UBA domain) and autophagosome receptor LC3 (via its LIR motif).\",\n      \"method\": \"ANKZF1 knockout, live-cell fluorescence imaging, co-immunoprecipitation, LIR mutagenesis, mitophagy flux assay\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — KO with functional readout, interaction validated by Co-IP, domain mutagenesis, single lab\",\n      \"pmids\": [\"40730577\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"The N-terminal 73 residues of human ANKZF1 negatively regulate its mitochondrial targeting by suppressing an internal matrix-targeting sequence-like sequence (iMTS-L) at residues 231–240. Deletion of the N-terminal segment causes structural rearrangement (shown by MD simulation) that exposes the 231–240 iMTS-L. Residues 231–324 constitute an independent mitochondrial signal that can target GFP to mitochondria when fused to its N-terminus.\",\n      \"method\": \"Truncation mutants, live-cell fluorescence localization, GFP-fusion targeting assay, molecular dynamics simulation\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiments with functional mutants and structural modeling, single lab\",\n      \"pmids\": [\"41920018\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"ANKZF1 interacts with YWHAE (14-3-3ε) to competitively inhibit cytoplasmic retention of YAP1, thereby promoting YAP1 nuclear import and transcriptional activation of pro-lymphangiogenic factors in clear-cell renal cell carcinoma. NAT10-mediated ac4C modification of ANKZF1 mRNA upregulates ANKZF1 expression.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, site-directed mutagenesis, RNA immunoprecipitation, mass spectrometry\",\n      \"journal\": \"Cancer communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — Co-IP with mechanistic follow-up in cancer cell context, single lab\",\n      \"pmids\": [\"38407929\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"ANKZF1 (human ortholog of yeast Vms1) is a peptidyl-tRNA hydrolase that cleaves the tRNA acceptor arm of ubiquitinated nascent chain–tRNA complexes on stalled 60S ribosomal subunits during ribosome-associated quality control (RQC), releasing proteasome-degradable ubiquitinated nascent chains; its mitochondrial targeting is autoinhibited by the N-terminal domain and relieved by stress signals, whereupon ANKZF1 antagonizes CAT-tailing to protect mitochondria from toxic polypeptide aggregation, and also participates in PINK1-Parkin-mediated mitophagy as an adaptor bridging ubiquitinated outer mitochondrial membrane proteins to LC3.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"ANKZF1 is a peptidyl-tRNA hydrolase central to ribosome-associated quality control (RQC) that cleaves the tRNA acceptor arm of ubiquitinated nascent chain–tRNA complexes on stalled 60S ribosomal subunits, releasing proteasome-degradable nascent chains [PMID:30244831, PMID:29632312]. Its catalytic mechanism depends on a conserved GSQ motif within the VLRF1 domain, which is positioned toward the CCA end of the P-site tRNA, with an ABCF-type ATPase (Arb1) stimulating cleavage in the pre-cleavage state [PMID:31189955]. At mitochondria, ANKZF1 antagonizes CAT-tailing of stalled nascent chains to prevent toxic polypeptide aggregation and sequestration of mitochondrial chaperones and respiratory chain subunits; its mitochondrial targeting is autoinhibited by the N-terminal domain, which masks an internal matrix-targeting sequence relieved by stress signals such as oxidized sterols [PMID:29107329, PMID:29149595, PMID:38670305, PMID:41920018]. ANKZF1 also functions as a mitophagy adaptor during PINK1-Parkin-mediated mitophagy, bridging polyubiquitinated outer mitochondrial membrane proteins (via its UBA domain) to LC3 on autophagosomes (via a LIR motif) [PMID:40730577].\",\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Establishing that Vms1/ANKZF1 cooperates with Cdc48/p97 in protein degradation pathways revealed its initial functional link to ubiquitin-proteasome quality control beyond a generic stress factor.\",\n      \"evidence\": \"Genetic deletion of VMS1 in yeast with Cdc13 degradation assay and epistasis analysis with autophagy mutants\",\n      \"pmids\": [\"22718752\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Substrate specificity beyond Cdc13 unknown\", \"Direct enzymatic activity not yet identified\", \"Mechanism of Vms1-Cdc48 cooperation unresolved\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Demonstrating that the N-terminal domain autoinhibits mitochondrial targeting of Vms1, with oxidative stress relieving this block, established the regulated translocation mechanism that controls ANKZF1 function at mitochondria.\",\n      \"evidence\": \"Truncation mutants and live-cell imaging with laser-induced mitochondrial ROS in yeast\",\n      \"pmids\": [\"23468520\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of the stress-induced signal relieving autoinhibition not determined\", \"Structural basis of intramolecular autoinhibition unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showing that the Cdc48-Vms1 complex is required for 26S proteasome assembly revealed an unexpected role in maintaining the proteolytic machinery itself, beyond being a simple substrate adaptor.\",\n      \"evidence\": \"VMS1 deletion in yeast with native gel analysis of proteasome complexes and ubiquitin accumulation assays\",\n      \"pmids\": [\"24351022\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect role in proteasome biogenesis unclear\", \"Not confirmed in mammalian cells\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Two concurrent studies resolved the core mitochondrial function of Vms1/ANKZF1: it binds 60S ribosomes at the mitochondrial surface to antagonize Rqc2-mediated CAT-tailing, preventing toxic aggregation of stalled nascent chains inside mitochondria, and its mitochondrial translocation is controlled by an autoinhibitory N-terminal–MTD interaction relieved by the oxidized sterol ergosterol peroxide.\",\n      \"evidence\": \"Crystal structure of MTD–sterol complex, competitive binding assays, genetic deletion with mitochondrial import and aggregation assays, ribosome fractionation in yeast\",\n      \"pmids\": [\"29107329\", \"29149595\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether identical sterol-based activation applies to human ANKZF1 is unresolved\", \"How Vms1 selects mitochondrial versus cytoplasmic stalled ribosomes unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identification of ANKZF1/Vms1 as a peptidyl-tRNA hydrolase that specifically cleaves the tRNA acceptor arm on ubiquitinated 60S RNCs defined its catalytic activity, showing it depends on a conserved glutamine (analogous to eRF1) and requires upstream NEMF/Listerin-dependent ubiquitination to render the substrate susceptible.\",\n      \"evidence\": \"In vitro reconstitution with purified components, active-site mutagenesis, tRNA cleavage mapping in both yeast and human systems\",\n      \"pmids\": [\"30244831\", \"29632312\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Kinetic parameters of the hydrolase activity uncharacterized\", \"Whether additional cofactors modulate activity in vivo unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Cryo-EM structures of Vms1 on the 60S subunit in pre- and post-cleavage states revealed how the VLRF1 domain positions its GSQ catalytic motif at the CCA end of P-site tRNA, how Y285 dislodges tRNA A73 to enable cleavage, and how the E-site ATPase Arb1 stimulates this reaction.\",\n      \"evidence\": \"Cryo-EM structure determination with functional mutagenesis and in vitro cleavage assays in yeast\",\n      \"pmids\": [\"31189955\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No equivalent high-resolution structure of human ANKZF1 on 60S\", \"Role of Arb1 ortholog in human RQC not tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Loss-of-function studies in human glioblastoma cells demonstrated that ANKZF1 depletion causes CAT-tail accumulation in mitochondria, sequestering chaperones (HSP60, mtHSP70) and respiratory chain subunits, activating the UPRmt and triggering mitochondrial apoptosis—providing the first direct evidence of ANKZF1's anti-CAT-tail function in human cells.\",\n      \"evidence\": \"siRNA knockdown with mitochondrial fractionation, co-IP, membrane potential assay, and xenograft model in human glioblastoma cells\",\n      \"pmids\": [\"38670305\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Findings from a single cancer cell type; generalizability to non-malignant cells untested\", \"Whether ANKZF1 acts catalytically or structurally to suppress CAT-tailing in human cells not resolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Discovery that ANKZF1 acts as a mitophagy adaptor during PINK1-Parkin-mediated mitophagy, bridging ubiquitinated outer mitochondrial membrane proteins to LC3 through its UBA domain and LIR motif (residues 333–336), expanded its function beyond RQC to organelle clearance.\",\n      \"evidence\": \"ANKZF1 knockout, co-IP of ANKZF1 with Parkin and LC3, LIR mutagenesis, and mitophagy flux assay in human cells\",\n      \"pmids\": [\"40730577\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding; independent confirmation needed\", \"Relative contribution of ANKZF1 versus other mitophagy receptors (OPTN, NDP52) not compared\", \"Whether RQC and mitophagy adaptor functions are coordinated or independent is unclear\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Mapping the human ANKZF1 N-terminal autoinhibition to residues 1–73 suppressing an internal matrix-targeting sequence at residues 231–240 provided the first structural rationale for regulated mitochondrial import of the human protein.\",\n      \"evidence\": \"Truncation mutants, GFP-fusion targeting assays, and molecular dynamics simulation in human cells\",\n      \"pmids\": [\"41920018\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"MD simulation-based structural model lacks experimental structural validation\", \"Endogenous signal that relieves N-terminal autoinhibition in human cells not identified\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key open questions remain: the identity of the physiological signal relieving ANKZF1 autoinhibition in human cells, a high-resolution structure of human ANKZF1 on the 60S subunit, whether the RQC peptidyl-tRNA hydrolase and mitophagy adaptor functions are coordinated, and the in vivo substrate spectrum of ANKZF1 beyond stalled nascent chains.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No high-resolution structure of human ANKZF1 on 60S ribosome\", \"Physiological activating signal in human cells unidentified\", \"Functional coordination between RQC and mitophagy roles untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [3, 4, 5, 8, 10]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005840\", \"supporting_discovery_ids\": [0, 1, 2, 3]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 2, 3, 8]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"complexes\": [\n      \"60S RQC complex\"\n    ],\n    \"partners\": [\n      \"NEMF\",\n      \"LTN1\",\n      \"VCP\",\n      \"PRKN\",\n      \"MAP1LC3B\",\n      \"YWHAE\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}