Affinage

ATE1

Arginyl-tRNA--protein transferase 1 · UniProt O95260

Length
518 aa
Mass
59.1 kDa
Annotated
2026-04-28
43 papers in source corpus 17 papers cited in narrative 17 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ATE1 encodes the arginyl-tRNA-protein transferase (R-transferase) that post-translationally conjugates arginine from Arg-tRNAArg to N-terminal acidic residues and internal side-chain carboxylates of substrate proteins, thereby creating Arg/N-degrons that target substrates for proteasomal degradation (PMID:2185248, PMID:24529990). Structural studies reveal a bilobed GNAT-fold enzyme whose active site resides at the domain–domain interface, with substrate and tRNA bound in adjacent pockets; the N-terminal domain harbors a regulatory [Fe-S] cluster that functions as an oxygen sensor, and hemin inhibits activity through disulfide-mediated oligomerization (PMID:35878037, PMID:36087779, PMID:40099869, PMID:37010764). ATE1-mediated arginylation regulates actin polymerization and focal adhesion formation, modulates myosin contractility in platelets, drives neuronal growth cone migration through co-translational arginylation of β-actin, and controls RGS protein turnover affecting retinal signaling and Wnt/β-catenin pathway activity (PMID:30586322, PMID:24293517, PMID:28844905, PMID:33931669, PMID:34158395). ATE1 alternative splicing of mutually exclusive exons is governed by competing intronic RNA secondary structures including an ultra-long-range interaction, and ATE1 protein levels increase under diverse stresses to promote arginylation-dependent cell death (PMID:33330934, PMID:27685622).

Mechanistic history

Synthesis pass · year-by-year structured walk · 11 steps
  1. 1990 High

    Establishing the molecular identity of ATE1 as the gene encoding arginyl-tRNA-protein transferase resolved how the N-end rule pathway activates degradation signals on protein substrates.

    Evidence Heterologous expression of yeast ATE1 in E. coli conferred Arg-transferase activity; ate1-null yeast lacked this activity and failed to degrade N-end rule substrates.

    PMID:2185248

    Open questions at the time
    • Substrate scope beyond N-end rule model peptides unknown
    • Mammalian ATE1 not yet characterized
    • Catalytic mechanism unresolved
  2. 2013 Medium

    Demonstrating that ATE1 directly interacts with myosin II and modulates its regulatory light chain phosphorylation extended arginylation beyond protein degradation to actomyosin contractility regulation.

    Evidence Platelet-specific Ate1 knockout mice showed enhanced MLC phosphorylation, increased clot retraction, and in vivo thrombus formation; Co-IP confirmed direct ATE1–myosin II interaction.

    PMID:24293517

    Open questions at the time
    • Whether myosin II is directly arginylated was not determined
    • Mechanism linking arginylation to MLC phosphorylation state unclear
    • Single Co-IP without reciprocal validation
  3. 2014 High

    Discovery of midchain arginylation at internal Asp/Glu side-chain carboxylates revealed that ATE1 catalytic scope extends beyond N-terminal modification, fundamentally broadening the substrate landscape.

    Evidence In vivo mass spectrometry identified internal arginylation sites on intact proteins; in vitro assays with purified ATE1 confirmed midchain conjugation.

    PMID:24529990

    Open questions at the time
    • Functional consequences of specific midchain arginylation events not established
    • Structural basis for midchain versus N-terminal selectivity unknown
  4. 2016 Medium

    Showing that ATE1 protein accumulates under diverse stresses and promotes arginylation-dependent cell death established ATE1 as a stress-response effector rather than solely a constitutive housekeeping enzyme.

    Evidence ATE1 KO/KD across yeast, mouse, and human cells reduced stress-induced cell death; complementation with catalytically inactive ATE1 failed to rescue.

    PMID:27685622

    Open questions at the time
    • Critical stress-induced arginylation substrates not identified
    • Mechanism of ATE1 protein stabilization under stress unknown
  5. 2017 Medium

    Localizing Ate1 mRNA to neuronal growth cones and showing co-translational arginylation of β-actin there connected ATE1 to local cytoskeletal remodeling required for neuronal migration and outgrowth.

    Evidence Conditional Ate1 KO in mouse nervous system caused growth cone defects, reduced neurite outgrowth, and decreased F-actin; FISH showed co-localization of Ate1 mRNA with arginylated β-actin at growth cone tips.

    PMID:28844905

    Open questions at the time
    • Zipcode elements in Ate1 mRNA not mapped
    • Whether arginylation of actin is sufficient to rescue growth cone defects not tested
  6. 2018 Medium

    Demonstrating that Ate1 knockout abolishes actin focal adhesions and reduces actin polymerization capacity in Dictyostelium confirmed a conserved role for arginylation in actin dynamics beyond vertebrates.

    Evidence Ate1-null Dictyostelium lacked focal adhesion sites; MS identified arginylation sites on actin; in vitro polymerization of ate1-null actin was diminished; Ate1-GFP relocated to actin-rich protrusions.

    PMID:30586322

    Open questions at the time
    • Whether individual arginylation sites on actin are functionally distinct not resolved
    • Direct structural effect of arginylation on actin filament properties unknown
  7. 2021 Medium

    Identifying RGS7 and RGS5 as ATE1-dependent degradation substrates connected arginylation to G-protein signaling modulation in retina and Wnt/β-catenin suppression in liver, respectively.

    Evidence Conditional Ate1 KO elevated RGS7 in retinal ON bipolar cells altering electroretinogram responses; ATE1 knockdown/overexpression in liver cancer cells showed RGS5 accumulation activating β-catenin via GSK3-β inhibition.

    PMID:33931669 PMID:34158395

    Open questions at the time
    • Whether RGS7/RGS5 are directly arginylated by ATE1 not shown
    • Generalizability of RGS family as ATE1 substrates not tested
  8. 2021 High

    Elucidating the RNA structural mechanism controlling ATE1 mutually exclusive exon splicing revealed how alternative ATE1 isoforms are generated through competing intronic secondary structures spanning 30 kb.

    Evidence Compensatory mutagenesis in minigenes and LNA/DNA mixmer blocking of endogenous pre-mRNA demonstrated R1–R4 competition for R3 base-pairing and ultra-long-range R2–R5 interaction.

    PMID:33330934

    Open questions at the time
    • Functional differences between exon 7a and 7b isoforms not biochemically defined
    • Regulation by RNA Pol II elongation rate not tested endogenously
  9. 2022 High

    Crystal structures of yeast and Kluyveromyces Ate1 resolved the bilobed GNAT-fold architecture, revealed the active site at the domain interface, and identified hemin inhibition and an [Fe-S] cluster regulatory domain, providing the first structural framework for the enzyme.

    Evidence X-ray crystallography of KlAte1 and ScAte1 with mutagenesis validation; hemin-induced oligomerization; SAXS and cryo-EM of apo ScAte1 showing N-terminal domain disorder.

    PMID:35878037 PMID:36087779

    Open questions at the time
    • No structure with both tRNA and substrate bound at this point
    • Mechanism by which [Fe-S] cluster senses oxygen not determined
    • Human enzyme structure not yet available
  10. 2023 Medium

    Biochemical reconstitution of the [Fe-S] cluster under anoxic conditions in both yeast and mouse ATE1 confirmed it as a bona fide oxygen-sensing cofactor regulating enzymatic activity.

    Evidence Anaerobic [Fe-S] cluster reconstitution with spectroscopic characterization of yeast and mouse ATE1.

    PMID:37010764

    Open questions at the time
    • Oxygen-dependent switching mechanism not structurally resolved
    • Physiological oxygen tension thresholds for ATE1 activity modulation unknown
  11. 2025 High

    The cryo-EM structure of human ATE1 bound to Arg-tRNAArg and an Nt-Asp peptide provided the first ternary complex view, explaining substrate selectivity through multivalent micromolar-affinity interactions and showing tRNA wrapping by an unstructured loop.

    Evidence Cryo-EM of human ATE1–Arg-tRNAArg–peptide complex with binding affinity measurements.

    PMID:40099869

    Open questions at the time
    • Catalytic mechanism (transition state) not captured
    • How midchain substrates are accommodated in the active site remains structurally unresolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • Key unresolved questions include the structural basis for midchain arginylation, the identity of critical stress-induced substrates mediating cell death, the biochemical distinction between ATE1 splice isoforms, and the physiological oxygen thresholds governing [Fe-S]-cluster-dependent activity regulation.
  • No structure of ATE1 with a midchain substrate
  • Stress-induced arginylation substrates driving apoptosis unidentified
  • Functional biochemistry of exon 7a vs 7b isoforms not compared

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016740 transferase activity 4 GO:0003723 RNA binding 2
Localization
GO:0005829 cytosol 3 GO:0005739 mitochondrion 1
Pathway
R-HSA-392499 Metabolism of proteins 4 GO:0005856 cytoskeleton 3 R-HSA-5357801 Programmed Cell Death 2 R-HSA-8953897 Cellular responses to stimuli 2 R-HSA-8953854 Metabolism of RNA 1

Evidence

Reading pass · 17 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1990 ATE1 encodes arginyl-tRNA-protein transferase (R-transferase) that catalyzes post-translational conjugation of Arg to N-terminal residues of acceptor proteins; expression of yeast ATE1 in E. coli (which lacks R-transferases) conferred Arg-transferase activity, and null ate1 mutants lack this activity and cannot degrade N-end rule substrates requiring Arg-transferase action. Heterologous expression in E. coli, null mutant analysis, in vitro enzymatic assay The Journal of biological chemistry High 2185248
2014 ATE1 catalyzes arginylation not only at N-terminal alpha-amino groups but also at internal side-chain carboxylates of Asp and Glu residues within intact proteins in vivo, revealing an unconventional midchain arginylation mechanism. Mass spectrometry (in vivo arginylation profiling), in vitro ATE1 arginylation assay with purified components Chemistry & biology High 24529990
2022 Crystal structure of K. lactis Ate1 R-transferase at 58 kDa reveals two domains that together recognize an acidic N-terminal residue of the acceptor substrate, the Arg residue of Arg-tRNAArg, and the 3'-proximal tRNAArg segment; the active site is located between the two domains. Hemin (Fe3+-heme) inhibits Nt-arginylation activity and induces disulfide-mediated oligomerization of Ate1. Site-directed mutagenesis guided by the structure identified specific binding residues. X-ray crystallography, site-directed mutagenesis, in vitro and in vivo arginylation assays Proceedings of the National Academy of Sciences of the United States of America High 35878037
2022 Crystal structure of S. cerevisiae ATE1 (ScATE1) in apo form reveals a bilobed protein with a GCN5-related N-acetyltransferase (GNAT) fold; the domain-domain interface constitutes the catalytic active site and tRNA-binding region. The N-terminal domain that binds a regulatory [Fe-S] cluster is dynamic and disordered in the absence of metal, indicating regulatory influence of this region. X-ray crystallography, SEC-SAXS, cryo-EM 2D class averaging, structural superposition Journal of molecular biology High 36087779
2025 Cryo-EM structure of human ATE1 in complex with Arg-tRNAArg and an N-terminal Asp peptide shows two adjacent pockets binding the Nt-substrate and Arg-tRNAArg respectively, the tRNA being wrapped by a long unstructured loop. In the apo state, two ATE1 monomers form a homodimer. Substrate selectivity is achieved through multivalent interactions with Kd values in the micromolar range. Cryo-EM structure determination, binding affinity measurements Autophagy High 40099869
2016 ATE1/arginylation promotes cell death and/or growth arrest in response to oxidative, heat, osmotic, heavy metal, and radiation stresses; ATE1 protein levels rise under acute stress, and the elevated ATE1 directly promotes cell death in an arginylation-activity-dependent manner. ATE1 is also required to suppress mutation frequency under DNA-damaging conditions. ATE1 knockout/knockdown in yeast, mouse, and human cells; genetic complementation with catalytically inactive ATE1; functional cell death and mutagenesis assays Cell death & disease Medium 27685622
2017 ATE1 localizes prominently to neuronal growth cones via zipcode-mediated mRNA targeting; Ate1 mRNA co-localizes with arginylated β-actin at growth cone tips, and co-translational arginylation of β-actin at growth cones drives growth cone migration and neurite outgrowth. Conditional Ate1 knockout in the nervous system causes defects in neuronal migration, reduced neurite outgrowth, and decreased doublecortin and F-actin in growth cones. Conditional mouse knockout (Nestin-Cre), live-cell imaging, FISH, fluorescence microscopy, protein synthesis inhibitor experiments Developmental biology Medium 28844905
2013 ATE1 directly interacts with myosin II in platelets (co-immunoprecipitation); platelet-specific ATE1 knockout leads to enhanced myosin regulatory light chain phosphorylation at Ser19, enhanced clot retraction, and increased in vivo thrombus formation, indicating arginylation modulates myosin contractility. Conditional platelet/megakaryocyte-specific knockout mouse, co-immunoprecipitation, phosphorylation analysis, in vivo thrombosis assay Haematologica Medium 24293517
2018 In Dictyostelium discoideum, Ate1 knockout abolishes focal actin adhesion sites and reduces adhesion; mass spectrometry identified four arginylation sites in the major actin isoform plus sites in actin-binding proteins. Actin from ate1-null cells has diminished polymerization capacity in vitro. Ate1-GFP rapidly relocalizes to sites of newly formed actin-rich protrusions. Gene knockout, live-cell microscopy, mass spectrometry, in vitro actin polymerization assay, chemotaxis assay Molecular biology of the cell Medium 30586322
2021 Ate1 regulates RGS7 protein levels by facilitating its proteasomal degradation; conditional Ate1 knockout in the nervous system elevates RGS7 levels in retinal ON bipolar cells, increases light-evoked ON-bipolar response sensitivities, and in cultured MEFs, RGS7 degradation by the proteasome is abolished in Ate1 knockout cells. Conditional mouse knockout (nervous system), electroretinography, proteasome inhibitor experiments in MEF cells Scientific reports Medium 33931669
2021 ATE1 splicing is controlled by competing RNA secondary structures: five conserved intronic elements (R1–R5) regulate mutual exclusion of exons 7a and 7b, with R1 and R4 competing for base-pairing with R3, and an ultra-long-range R2–R5 interaction spanning 30 kb. Disruption of these RNA structures by mutation or LNA/DNA mixmers abolishes mutually exclusive exon splicing, and co-transcriptional folding links exon 7a inclusion to RNA Pol II elongation rate. Minigene splicing assay with single/double/compensatory mutations, LNA/DNA mixmer blocking, endogenous pre-mRNA analysis Nucleic acids research High 33330934
2020 Liat1 (Ligand of Ate1) physically interacts with Ate1 and undergoes liquid-liquid phase separation in the nucleolus via an intrinsically disordered N-terminal region; Jmjd6 lysyl-hydroxylase modifies Liat1's poly-K region and inhibits its nucleolar targeting. Bimolecular fluorescence complementation, immunocytochemistry, IDR characterization, phase separation assay Proceedings of the National Academy of Sciences of the United States of America Medium 33443146
2023 ATE1 binds a regulatory iron-sulfur ([Fe-S]) cluster in its N-terminal domain that functions as an oxygen sensor to regulate ATE1 activity; chemical reconstitution under anoxic conditions restores [Fe-S] cluster binding in both yeast and mouse ATE1. Anaerobic [Fe-S] cluster reconstitution, spectroscopic characterization Methods in molecular biology Medium 37010764
2024 Mouse ATE1 contains an intrinsically disordered region (IDR) absent in yeast ATE1; HDX-MS and SAXS demonstrate this IDR is present in all mouse ATE1 splice variants and is predicted to facilitate complex formation with tRNAArg. SEC, SAXS, HDX-MS, AlphaFold modeling, bioinformatics Biochemistry Medium 39642180
2024 tRNAArg directly binds in vitro to TDP-43 RNA recognition motifs and stabilizes binding via TDP-43 dimerization (promoted by the N-terminal domain and NLS); the arginylation cofactor LIAT1 also binds tRNAArg in vitro, identifying LIAT1 as an RNA-binding protein. In vitro binding assays with recombinant proteins and in vitro-transcribed tRNA microPublication biology Low 39081859
2021 ATE1 inhibits liver cancer growth by facilitating proteasomal degradation of RGS5, which in turn maintains GSK3-β activity and promotes β-catenin degradation, thereby suppressing Wnt/β-catenin signaling; loss- and gain-of-function assays with RGS5 and GSK3-β inhibitor confirmed RGS5 as a key effector downstream of ATE1. Lentivirus-mediated knockdown/overexpression, loss- and gain-of-function, GSK3-β inhibitor rescue experiments, in vitro and in vivo tumor assays Molecular cancer research Medium 34158395
2024 Mitochondrial translocation of Ate1 is promoted by oxidative stressors and is essential for inducing apoptotic cell death; Ate1-induced cell death depends on mitochondrial permeability pore formation and apoptosis-inducing factor, but not on electron transport chain activity or ROS, and is independent of cytosolic ubiquitin-proteasome, autophagy, or ER stress pathways. Yeast genetic and cell biology experiments, mitochondrial fractionation, mitochondrial permeability pore assays, AIF analysis bioRxivpreprint Medium bio_10.1101_2024.11.22.624728

Source papers

Stage 0 corpus · 43 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1990 Cloning and functional analysis of the arginyl-tRNA-protein transferase gene ATE1 of Saccharomyces cerevisiae. The Journal of biological chemistry 140 2185248
2014 Arginyltransferase ATE1 catalyzes midchain arginylation of proteins at side chain carboxylates in vivo. Chemistry & biology 77 24529990
2016 Posttranslational arginylation enzyme Ate1 affects DNA mutagenesis by regulating stress response. Cell death & disease 38 27685622
2017 Arginyltransferase ATE1 is targeted to the neuronal growth cones and regulates neurite outgrowth during brain development. Developmental biology 34 28844905
2013 Loss of ATE1-mediated arginylation leads to impaired platelet myosin phosphorylation, clot retraction, and in vivo thrombosis formation. Haematologica 27 24293517
2022 Crystal structure of the Ate1 arginyl-tRNA-protein transferase and arginylation of N-degron substrates. Proceedings of the National Academy of Sciences of the United States of America 24 35878037
2021 Multiple competing RNA structures dynamically control alternative splicing in the human ATE1 gene. Nucleic acids research 24 33330934
2020 ATE1-Mediated Post-Translational Arginylation Is an Essential Regulator of Eukaryotic Cellular Homeostasis. ACS chemical biology 24 33228359
2021 ATE1 Inhibits Liver Cancer Progression through RGS5-Mediated Suppression of Wnt/β-Catenin Signaling. Molecular cancer research : MCR 19 34158395
1991 Physical, transcriptional and genetical mapping of a 24 kb DNA fragment located between the PMA1 and ATE1 loci on chromosome VII from Saccharomyces cerevisiae. Yeast (Chichester, England) 17 1882552
2021 The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1. Journal of molecular biology 16 34896361
2018 Ate1-mediated posttranslational arginylation affects substrate adhesion and cell migration in Dictyostelium discoideum. Molecular biology of the cell 16 30586322
2022 The Structure of Saccharomyces cerevisiae Arginyltransferase 1 (ATE1). Journal of molecular biology 9 36087779
2021 Arginyltransferase (Ate1) regulates the RGS7 protein level and the sensitivity of light-evoked ON-bipolar responses. Scientific reports 9 33931669
2025 An unbiased proteomic platform for ATE1-based arginylation profiling. Nature chemical biology 6 40855110
2020 The Ligand of Ate1 is intrinsically disordered and participates in nucleolar phase separation regulated by Jumonji Domain Containing 6. Proceedings of the National Academy of Sciences of the United States of America 6 33443146
2013 Disruption of the ATE1 and SLC12A1 Genes by Balanced Translocation in a Boy with Non-Syndromic Hearing Loss. Molecular syndromology 6 24550759
2022 Arginyl-tRNA-protein transferase 1 (ATE1) promotes melanoma cell growth and migration. FEBS letters 5 35561126
2015 Assaying ATE1 Activity In Vitro. Methods in molecular biology (Clifton, N.J.) 4 26285883
2025 An Unbiased Proteomic Platform for ATE1-based Arginylation Profiling. bioRxiv : the preprint server for biology 3 38854050
2024 tRNA Arg binds in vitro TDP-43 RNA recognition motifs and ligand of Ate1 protein LIAT1. microPublication biology 3 39081859
2022 Liquiritin Attenuates Angiotensin II-Induced Cardiomyocyte Hypertrophy via ATE1/TAK1-JNK1/2 Pathway. Evidence-based complementary and alternative medicine : eCAM 3 35341136
2015 Bacterial Expression and Purification of Recombinant Arginyltransferase (ATE1) and Arg-tRNA Synthetase (RRS) for Arginylation Assays. Methods in molecular biology (Clifton, N.J.) 3 26285882
2025 The structure-function relationship of ATE1 R-transferase of the autophagic Arg/N-degron pathway. Autophagy 2 40099869
2025 ATE1 promotes breast cancer progression via arginylation-dependent regulation of MAPK-MYC signaling. Cell communication and signaling : CCS 2 40898325
2024 Identification of an Intrinsically Disordered Region (IDR) in Arginyltransferase 1 (ATE1). Biochemistry 2 39642180
2015 Correlated Measurement of Endogenous ATE1 Activity on Native Acceptor Proteins in Tissues and Cultured Cells to Detect Cellular Aging. Methods in molecular biology (Clifton, N.J.) 2 26285879
2015 Assaying ATE1 Activity in Yeast by β-Gal Degradation. Methods in molecular biology (Clifton, N.J.) 2 26285881
2025 Method Overview for Discovering ATE1 Substrates and their Arginylation Sites. Chembiochem : a European journal of chemical biology 1 41147131
2024 Arginyltransferase 1 (ATE1)-mediated proteasomal degradation of viral haemagglutinin protein: a unique host defence mechanism. The Journal of general virology 1 39207120
2023 Reconstitution of the Arginyltransferase (ATE1) Iron-Sulfur Cluster. Methods in molecular biology (Clifton, N.J.) 1 37010764
2023 Structural analysis of human ATE1 isoforms and their interactions with Arg-tRNAArg. Journal of biomolecular structure & dynamics 1 37505085
2022 The preparation of recombinant arginyltransferase 1 (ATE1) for biophysical characterization. Methods in enzymology 1 36682863
2025 Erratum: Corrigendum: tRNA Arg binds in vitro TDP-43 RNA recognition motifs and ligand of Ate1 protein LIAT1. microPublication biology 0 39897169
2025 Cell based high-throughput screening for small molecule inhibitors of ATE1. SLAS discovery : advancing life sciences R & D 0 40816642
2025 Recombinant expression, purification, and characterization of human ATE1 arginyltransferase. Methods in enzymology 0 40887163
2024 Identification of an intrinsically disordered region (IDR) in arginyltransferase 1 (ATE1). bioRxiv : the preprint server for biology 0 39229138
2023 Preparation of ATE1 Enzyme from Native Mammalian Tissues. Methods in molecular biology (Clifton, N.J.) 0 37010746
2023 Correlated Measurement of Endogenous ATE1 Activity on Native Acceptor Proteins in Tissues and Cultured Cells to Detect Cellular Aging. Methods in molecular biology (Clifton, N.J.) 0 37010747
2023 Assaying ATE1 Activity in Yeast by β-Gal Degradation. Methods in molecular biology (Clifton, N.J.) 0 37010749
2023 Bacterial Expression and Purification of Recombinant Arginyltransferase (ATE1) and Arg-tRNA Synthetase (RRS) for Arginylation Assays. Methods in molecular biology (Clifton, N.J.) 0 37010752
2023 Assaying ATE1 Activity In Vitro. Methods in molecular biology (Clifton, N.J.) 0 37010756
2015 Preparation of ATE1 Enzyme from Native Mammalian Tissues. Methods in molecular biology (Clifton, N.J.) 0 26285878