Affinage

Showing GET3 — TRC40 is a alias.

GET3

ATPase GET3 · UniProt O43681

Length
348 aa
Mass
38.8 kDa
Annotated
2026-06-10
53 papers in source corpus 39 papers cited in narrative 40 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 8/8 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

GET3 (TRC40/ASNA1) is a conserved homodimeric ATPase that serves as the central cytosolic targeting factor of the GET/TRC pathway for post-translational delivery of tail-anchored (TA) membrane proteins to the ER (PMID:25745174, PMID:20375064). Nucleotide state drives an open-to-closed conformational cycle: in the closed, ADP·AlF4−-bound state the dimer forms a large hydrophobic groove spanning the interface that captures and shields the TA transmembrane helix, while the open state disrupts this groove to release substrate (PMID:19675567, PMID:19956640, PMID:35851188). A single TA protein binds one Get3 homodimer, with the α-helical/finger subdomain forming the binding surface (PMID:25745174, PMID:19948960). Get3 is primed for substrate capture by the upstream Get4–Get5 complex, which interacts with Get3 in a nucleotide-dependent manner, promotes the ATP-bound configuration optimal for capture, and remodels the lateral walls of the binding chamber to facilitate hand-off of TA substrate from the cochaperone Sgt2 (PMID:20554915, PMID:24727835, PMID:40902977). Loaded Get3 docks onto the ER membrane receptor — Get1/Get2 in yeast, WRB/CAML in mammals — which remodels Get3 toward the open state to drive insertion; ATP hydrolysis promotes efficient insertion and Get3 release from the receptor (PMID:21719644, PMID:19948960, PMID:21444755, PMID:22684149, PMID:27226539). Substrates extend beyond classical TA proteins to Golgi-resident TA proteins, SNAREs, VAPA/VAPB, the inner nuclear membrane protein emerin, short secretory precursors, and the anti-apoptotic protein MCL1 (PMID:22505607, PMID:26675233, PMID:31182645, PMID:42098326). Independently of its targeting role, Get3 acts as an ATP-independent holdase chaperone: oxidative stress triggers nucleotide-controlled disulfide formation at conserved cysteines, metal release, and assembly into higher-order chaperone-active oligomers that protect against protein aggregation, a state mutually exclusive with targeting and reversed by reduction and ATP rebinding (PMID:25242142, PMID:35839781). In mammals ASNA1 is essential for early embryogenesis (PMID:16797549), and TRC40-mediated insertion of emerin links the pathway to Emery-Dreifuss muscular dystrophy, as EDMD-causing emerin mutations disrupt TRC40 binding and membrane integration (PMID:26675233).

Mechanistic history

Synthesis pass · year-by-year structured walk · 15 steps
  1. 2006 Medium

    Before the GET pathway was defined, it was unknown how Get3 connected physically and genetically to the ER membrane machinery; this established Get3 as a partner of the Get1/Get2 transmembrane proteins.

    Evidence Co-immunoprecipitation and yeast genetic epistasis (suppressor and double-mutant analysis)

    PMID:16816426

    Open questions at the time
    • No structural basis for the interaction
    • Mechanism of TA insertion not yet addressed
    • Predates identification of TA proteins as substrates
  2. 2006 Medium

    The physiological requirement for the mammalian ortholog was unknown; knockout established Asna1 as essential for early mammalian development.

    Evidence Homologous recombination Asna1 knockout mouse with embryonic lethality analysis

    PMID:16797549

    Open questions at the time
    • No molecular mechanism for lethality identified
    • Does not distinguish targeting from chaperone roles
  3. 2007 Medium

    It was unclear whether ASNA1 had organismal physiological roles beyond a housekeeping function; this revealed a conserved nonautonomous role in insulin secretion.

    Evidence C. elegans loss- and gain-of-function genetics with insulin secretion readouts plus human beta-cell knockdown/overexpression

    PMID:17289575

    Open questions at the time
    • Mechanistic link between TA targeting and insulin secretion unresolved
    • Direct secretory substrates not identified at this stage
  4. 2009 High

    How Get3 binds and releases TA substrates was unknown; structures of open and closed dimer states defined a nucleotide-regulated hydrophobic groove at the dimer interface as the TA-binding site.

    Evidence X-ray crystallography of open (nucleotide-free) and closed (ADP·AlF4−) states with mutagenesis of groove residues, across multiple yeast/fungal/archaeal orthologs and H/D exchange MS

    PMID:19675567 PMID:19706470 PMID:19948960 PMID:19956640 PMID:20015340

    Open questions at the time
    • Stoichiometry of the functional Get3–TA complex not yet defined
    • Direct visualization of a bound TA helix lacking
  5. 2010 High

    Whether Get3 alone could insert TA proteins and what upstream factors regulated it were open; reconstitution showed receptor- and nucleotide-dependent insertion, and Get4/5 was placed upstream of Get3.

    Evidence In vitro reconstitution of TA insertion from recombinant components and ER membranes, plus crystallography and nucleotide-dependent binding assays of the Get4/5 complex

    PMID:20106980 PMID:20375064 PMID:20554915

    Open questions at the time
    • Stoichiometry of Get3–Get4/5 assembly unresolved
    • Mechanism of Sgt2-to-Get3 hand-off not yet defined
  6. 2011 High

    The identity of the mammalian receptor and how the receptor remodels Get3 were unknown; WRB was identified as the mammalian receptor and Get1 structures showed how it stabilizes the open Get3 state.

    Evidence Crystallography of Get3–Get1/Get2 complexes, biophysical (ITC/SAXS) stoichiometry of Get3–Get4/5, and biochemical/imaging identification of WRB

    PMID:21444755 PMID:21719644 PMID:22190685 PMID:22684149

    Open questions at the time
    • Full mammalian receptor composition not yet established
    • Order of conformational events during insertion incompletely defined
  7. 2014 High

    How Get4 regulates Get3's catalytic cycle was unresolved; the ternary Get3–Get4–Get5 structure showed Get4 primes Get3 into the optimal substrate-capture configuration and regulates ATP hydrolysis.

    Evidence X-ray crystallography of the ATP-bound ternary complex with structure-guided mutagenesis and TA targeting assays

    PMID:24727835

    Open questions at the time
    • Lateral transfer geometry from Sgt2 not yet visualized
    • Coupling of priming to nucleotide turnover incompletely defined
  8. 2014 High

    Whether Get3 had functions beyond targeting was unknown; oxidative stress was shown to convert it into an ATP-independent holdase chaperone via disulfide formation and oligomerization, mutually exclusive with targeting.

    Evidence In vitro chaperone assays, disulfide-disrupting mutagenesis, energy-depletion imaging, and yeast oxidative-stress phenotypes

    PMID:23203805 PMID:25242142

    Open questions at the time
    • Ordered redox activation steps not yet defined
    • Client spectrum of the chaperone state unresolved
  9. 2015 High

    The composition of the functional targeting complex and the physiological substrate range were unclear; reconstitution fixed the 1 TA : 1 Get3 dimer stoichiometry and crystal structures resolved TA-helix recognition, while emerin established disease relevance.

    Evidence In vitro reconstitution plus crystallography of Get3–TA complexes; proximity ligation, in vitro insertion, and transport assays for emerin

    PMID:25745174 PMID:26675233

    Open questions at the time
    • Full client repertoire still being defined
    • Mechanism linking emerin mistargeting to muscular dystrophy pathology not detailed
  10. 2016 High

    The minimal mammalian receptor was undefined; reconstitution showed WRB and CAML together are sufficient and necessary for TRC40-mediated insertion, with CAML in molar excess and mutual level regulation.

    Evidence Functional proteoliposome reconstitution from detergent extract with immunodepletion, in vitro synthesized components, and quantitative Western blot

    PMID:27226539

    Open questions at the time
    • Stoichiometric architecture of the WRB/CAML receptor in membranes unresolved
    • Mechanism of mutual level regulation not defined
  11. 2019 High

    The full conformational cycle and substrate spectrum of the targeting factor were incompletely resolved; cryo-EM across five states and dominant-negative proteomics defined nucleotide-driven CBD reorganization and broadened the client list to Golgi TA proteins and VAPA/VAPB.

    Evidence Cryo-EM of Get3 in five nucleotide/conformational states; dominant-negative TRC40(D74E) trap with quantitative mass spectrometry; CRISPR-based Retro-2 target validation

    PMID:31182645 PMID:31674906 PMID:35851188

    Open questions at the time
    • Substrate selectivity rules not fully defined
    • Some substrates persist despite groove mutation, implying additional binding determinants
  12. 2022 High

    How the redox switch is ordered and reset was unknown; the activation cycle was resolved as nucleotide-controlled cysteine reactivity, oxidation-induced unfolding into chaperone oligomers, and reduction-plus-ATP-driven inactivation enabling client hand-off.

    Evidence Biochemical redox and chaperone assays, conserved-cysteine mutagenesis, and yeast oxidative-stress genetics

    PMID:35839781

    Open questions at the time
    • Identity of downstream ATP-dependent chaperones receiving clients not defined
    • Structural basis of the chaperone-active oligomer unresolved
  13. 2025 High

    How Get4/5 enables lateral TA hand-off from Sgt2 was unresolved; the cryo-EM Get3–Get4/5 structure showed remodeling of a 'lateral gate' that opens the binding chamber adjacent to the Sgt2-binding domain of Get5.

    Evidence Cryo-EM, molecular dynamics, mutagenesis, ATPase and binding assays of the S. cerevisiae Get3–Get4/5 complex

    PMID:40902977

    Open questions at the time
    • A captured Sgt2–Get3 transfer intermediate not yet visualized
    • Timing of lateral gate opening relative to nucleotide turnover unresolved
  14. 2025 Medium

    The tissue-specific physiological consequences of losing the targeting machinery were unclear; cardiomyocyte- and neuron-specific knockouts showed cell-autonomous requirements linked to TA substrate destabilization and impaired membrane trafficking.

    Evidence Constitutive and inducible conditional knockout mouse models with substrate expression analysis, transcriptomics, and spinal cord histology

    PMID:39823474 PMID:41370295

    Open questions at the time
    • Specific TA substrates driving each tissue phenotype not pinpointed
    • Relative contribution of targeting versus chaperone roles to phenotypes unresolved
  15. 2026 Medium

    Whether GET3 regulates apoptotic machinery as cargo was unknown; MCL1 was identified as a TA-containing client whose levels and pro-survival function depend on GET3.

    Evidence Degron-mediated GET3 depletion, co-immunoprecipitation, apoptosis and clonogenic survival assays

    PMID:42098326

    Open questions at the time
    • Whether MCL1 is inserted via the canonical WRB/CAML receptor not shown
    • In vivo relevance of GET3–MCL1 axis to tumor survival unresolved

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the cell switches Get3 between its targeting and holdase chaperone modes in vivo, and which downstream chaperones and physiological substrates govern recovery from stress, remains to be defined.
  • No structure of the chaperone-active oligomer
  • Substrate hand-off partners during stress recovery not identified
  • Quantitative balance between targeting and chaperone pools in tissues unknown

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140657 ATP-dependent activity 5 GO:0044183 protein folding chaperone 3 GO:0140104 molecular carrier activity 3 GO:0140299 molecular sensor activity 3 GO:0008289 lipid binding 2
Localization
GO:0005783 endoplasmic reticulum 4 GO:0005829 cytosol 3
Pathway
R-HSA-9609507 Protein localization 4 R-HSA-392499 Metabolism of proteins 3 R-HSA-5653656 Vesicle-mediated transport 2 R-HSA-8953897 Cellular responses to stimuli 2
Partners
CAMLGET1GET2GET4GET5SGT2VAPBWRB
Complex memberships
Get3-Get1/Get2 receptor complexGet3-Get4-Get5 pre-targeting complexTRC40-WRB/CAML receptor complex

Evidence

Reading pass · 40 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2009 Crystal structures of yeast Get3 in 'open' (nucleotide-free) and 'closed' (ADP·AlF4−-bound) dimer states revealed that in the closed state the dimer interface contains a large hydrophobic groove responsible for tail-anchored protein binding, and in the open state this groove is disrupted; mutational analyses confirmed the groove's role in TA protein binding, establishing a nucleotide-regulated binding/release mechanism. X-ray crystallography (open and closed dimer states) + site-directed mutagenesis of hydrophobic groove residues Nature High 19675567
2015 Reconstitution of the physiological assembly pathway showed that the functional Get3–TA protein targeting complex comprises a single TA protein bound to a Get3 homodimer; crystal structures of Get3 bound to different TA proteins showed the α-helical transmembrane domain occupying a hydrophobic groove spanning the homodimer interface, elucidating the mechanism of TA protein recognition and shielding. In vitro reconstitution of targeting complex + X-ray crystallography of Get3–TA protein complexes with different substrates Science High 25745174
2011 Crystal structures of Get3 in complex with cytosolic domains of the ER membrane receptor subunits Get1 and Get2 showed that Get1 and Get2 use adjacent, partially overlapping binding sites on Get3 and can bind simultaneously; docking to the Get1/2 receptor induces conformational changes in Get3 required for TA protein insertion. X-ray crystallography of Get3–Get1/2 receptor complexes at 3.0, 3.2, and 4.6 Å + biochemical experiments Science High 21719644
2009 Structural and biochemical analyses of Get3 (Asna1/TRC40) showed that the α-helical subdomain binds the TA protein transmembrane domain; amide proton exchange mass spectrometry mapped the TA-binding site to the α-helical subdomain; in vitro membrane insertion assays demonstrated Get3 inserts Ramp4 in a nucleotide- and receptor-dependent manner; ATP hydrolysis is not strictly required for insertion but is needed for efficient insertion, likely for Get3 release from its receptor. X-ray crystallography + hydrogen/deuterium exchange MS + in vitro membrane insertion reconstitution assay Proceedings of the National Academy of Sciences of the United States of America High 19948960
2009 Crystal structures of Get3 from S. cerevisiae (apo, open) and D. hansenii (ADP-bound, closed) identified a nucleotide-binding domain and a 'finger' domain with a hydrophobic groove as the TA protein TMD-binding site; a hydrophobic helix from a symmetry-related molecule occupying the groove mimicked TA binding, and the open/closed conformational switch is linked to TA protein release. X-ray crystallography of Get3 from two yeast species in apo and ADP-bound states PloS one High 19956640
2009 Crystal structures of apo and ADP-bound Get3 from S. cerevisiae and A. fumigatus identified residues important for dimer interfaces; structure-guided mutagenesis confirmed key interfaces and essential residues coupling ATP hydrolysis to TA protein binding and release. X-ray crystallography (apo and ADP-bound forms, two species) + structure-guided mutagenesis Proceedings of the National Academy of Sciences of the United States of America High 19706470
2009 Crystal structures of Get3 in ADP-bound and nucleotide-free forms showed a Zn2+-mediated homodimer in head-to-head orientation; cross-linking experiments indicated the closed dimer stimulates ATP hydrolysis; coexpression-based binding assays demonstrated direct interaction between the helical domain of Get3 and the Sec22p TMD independent of ATP and dimer formation; the conserved DTAPTGH motif was proposed to link ATP hydrolysis to TA protein insertion. X-ray crystallography + chemical cross-linking + coexpression-based binding assay Genes to cells Medium 20015340
2010 Get3 (Asna1/TRC40) was shown to mediate membrane insertion of the TA proteins RAMP4 and Sec61β from recombinant Asna1–TA protein complexes into ER-derived membranes in a mechanism requiring ATP or ADP and a protease-sensitive ER membrane receptor, but not additional cytosolic factors; cytochrome b5 insertion proceeded independently of Asna1 and nucleotides. In vitro reconstitution of TA protein membrane insertion from recombinant components + ER-derived membranes + protease sensitivity assay Journal of cell science High 20375064
2011 WRB (tryptophan-rich basic protein/CHD5), an ER-resident membrane protein with sequence similarity to yeast Get1, was identified as the ER membrane receptor for mammalian TRC40/Asna1; the coiled-coil domain of WRB is the binding site for TRC40; a soluble coiled-coil domain fragment interfered with TRC40-mediated TA protein membrane insertion. Biochemical interaction assays + cell imaging + dominant-negative soluble fragment inhibition of in vitro TA insertion Journal of cell science High 21444755
2010 Crystal structure of Get4 with an N-terminal Get5 fragment showed they form an intimate complex existing as a dimer-of-heterodimers; Get3 binds to a conserved surface on Get4 in a nucleotide-dependent manner, placing Get4/5 upstream of Get3 in the TA protein delivery pathway. X-ray crystallography of Get4/5 complex + nucleotide-dependent binding assay of Get3 to Get4 Proceedings of the National Academy of Sciences of the United States of America High 20554915
2010 Co-immunoprecipitation and crystallographic studies showed Get4 and Get5 form a tight complex; Get3 physically and transiently interacts with the Get4–Get5 complex; Sgt2 interacts with Get5; genetic interactions between GET3, GET4, GET5, and the chaperone YDJ1 implicate molecular chaperones in TA protein insertion. Co-immunoprecipitation + X-ray crystallography of Get4/5 + genetic epistasis (YDJ1) The Journal of biological chemistry Medium 20106980
2011 ITC and SAXS demonstrated that the Get3 homodimer interacts with two copies of the Get4–Get5 complex to form an extended stoichiometric complex in solution, defining the interaction surface as dominated by electrostatic forces. Isothermal titration calorimetry (ITC) + small-angle X-ray scattering (SAXS) The Journal of biological chemistry Medium 22190685
2011 Crystal structures of ADP-bound Get3 in complex with the cytoplasmic domain of Get1 in open and semi-open conformations showed that Get1 uses two distinct interfaces to bind Get3 and stabilize its open dimer conformation, one sufficient for binding and the second required to stabilize the open state. X-ray crystallography (3.0 and 4.5 Å) of Get3–Get1CD complexes + biochemical binding assays Journal of molecular biology High 22684149
2011 An archaeal Get3 homologue crystallized as a tetramer; SAXS of a fungal Get3–TA protein complex showed a molecular envelope consistent with the archaeal tetramer; the tetramer generates a hydrophobic chamber that binds TA proteins; the fungal tetramer complex was capable of mediating TA insertion in vitro. X-ray crystallography of archaeal Get3 tetramer + SAXS of fungal Get3–TA complex + in vitro TA insertion assay The EMBO journal Medium 22122436
2014 Get3 switches to an ATP-independent chaperone function upon oxidative stress: oxidation causes disulfide bond formation, metal release, and formation of distinct higher-order oligomeric structures; this chaperone activity is functionally distinct from and mutually exclusive with its TA protein targeting function; yeast cells lacking Get3 show oxidative stress-sensitive phenotypes attributable to loss of this chaperone activity. In vitro chaperone activity assays + mutational analysis (disulfide-disrupting mutants) + yeast genetic phenotypic analysis under oxidative stress Molecular cell High 25242142
2012 Get3 functions as an ATP-independent holdase chaperone during energy depletion (glucose starvation), reversibly relocating to deposition sites for protein aggregates (alongside generic chaperones Hsp42, Ssa2, Sis1, Hsp104) to sequester TA proteins under conditions preventing their membrane insertion. Live-cell fluorescence imaging of Get3-GFP localization + genetic and cell biological analyses under energy depletion Journal of cell science Medium 23203805
2014 Crystal structure of a yeast Get3–Get4–Get5 complex in an ATP-bound state showed how Get4 primes Get3 by promoting the optimal configuration for substrate capture; structure-guided biochemical analyses demonstrated that Get4-mediated regulation of Get3's ATP hydrolysis is essential for efficient TA protein targeting. X-ray crystallography of Get3–Get4–Get5 ternary complex + structure-guided mutagenesis + biochemical TA targeting assays Nature structural & molecular biology High 24727835
2012 TRC40 (Get3) binds short secretory protein precursors (apelin, statherin, preprocecropin A) and can deliver them to the ER membrane for post-translational translocation via the Sec61 translocon, identifying secretory proteins as a class of TRC40 substrates beyond TA proteins. In vitro TRC40-binding assay + in vitro ER translocation assay with dominant-negative TRC40 inhibition Journal of cell science Medium 22505607
2013 TRC40 (Get3) is dispensable for peroxisomal targeting of the TA protein PEX26; PEX19 captures PEX26 in the cytosol and delivers it directly to peroxisomes via PEX3, defining a TRC40-independent class I pathway for peroxisomal TA proteins and showing that basic residues in PEX26's luminal domain are essential for PEX19 binding. Coimmunoprecipitation of PEX19–PEX26 complex + dominant-negative TRC40 inhibition assay + mutagenesis of PEX26 luminal basic residues The Journal of cell biology Medium 23460677
2011 TRC40/Asna1 mediates post-translational membrane insertion of calneuron-1 and calneuron-2 (neuronal TA calcium sensors) via their 23-amino-acid TMD; calneuron dimerization/multimerization via the TMD precludes TRC40 binding and membrane insertion, but no cytosolic pool of calneurons was detected, indicating self-association is restricted to membrane-inserted protein. In vitro TRC40 binding assay + co-immunoprecipitation + in vitro membrane insertion assay The Journal of biological chemistry Medium 21878631
2015 TRC40-mediated TA protein insertion is required for correct trafficking of emerin to the inner nuclear membrane; emerin interacts with TRC40 in situ (proximity ligation assay), is inserted into microsomal membranes in an ATP- and TRC40-dependent manner, and EDMD-causing emerin mutations disrupt TRC40 binding, membrane integration, and inner nuclear membrane targeting. Proximity ligation assay + in vitro microsomal membrane insertion assay + rapamycin-based dimerization transport assay + dominant-negative WRB/CAML receptor fragments Journal of cell science High 26675233
2016 Functional proteoliposomes reconstituted from microsomal detergent extracts lost TA protein insertion activity when depleted of TRC40-associated proteins or CAML itself; in vitro synthesized CAML and WRB together were sufficient to confer insertion competence to liposomes, defining the minimal mammalian receptor for TRC40-mediated TA insertion; CAML is present in ~5-fold molar excess over WRB and they mutually regulate each other's levels. Reconstituted functional proteoliposomes from detergent extract + immunodepletion + in vitro insertion assay + quantitative Western blot The Journal of biological chemistry High 27226539
2019 CAML, in the presence of sufficient WRB, is inserted into the ER membrane with three transmembrane segments in its C-terminal region; without sufficient WRB, CAML fails to adopt correct topology, generating aberrant topoforms that aggregate at ER-associated clusters and are degraded by the proteasome; WRB acts catalytically to assist CAML topogenesis. Topology mapping assays + proteasome inhibitor experiments + ER localization imaging Scientific reports Medium 31417168
2019 The small molecule Retro-2 blocks delivery of newly synthesized TA proteins to the ER-targeting factor ASNA1 (TRC40); a single ASNA1 point mutant identified by CRISPR-mediated mutagenesis abolishes both Retro-2's cytoprotective effect against ricin and its inhibitory effect on ASNA1-mediated ER targeting, demonstrating that Retro-2 acts directly on the ASNA1-dependent TA protein targeting step to prevent retrograde trafficking of ricin. CRISPRi genetic interaction screen + cell-based TA protein targeting assay + in vitro ASNA1 interaction assay + CRISPR point mutant resistance mapping eLife High 31674906
2019 A dominant-negative ATPase-impaired TRC40 mutant (D74E) traps TA protein clients in the cytoplasm; manipulation of the hydrophobic TA-binding groove reduces interaction with most but not all substrates; identified known and novel TRC40 substrates including Golgi-resident TA proteins (golgin-84, CASP, giantin) and VAPA/VAPB by quantitative mass spectrometry. Dominant-negative TRC40(D74E) trap + quantitative mass spectrometry + groove mutant analysis Journal of cell science Medium 31182645
2022 Activation of Get3's chaperone function follows a multi-step process: reactivity of two conserved cysteines is directly controlled by Get3's nucleotide-binding state; thiol oxidation causes local unfolding and transition into chaperone-active oligomers; inactivation requires cysteine reduction followed by ATP binding, enabling transfer of client proteins to ATP-dependent chaperones for refolding; disrupting this cycle in yeast impairs oxidative stress resistance. Biochemical redox assays + mutagenesis of conserved cysteines + in vitro chaperone activity assays + yeast genetic phenotypic analyses Molecular cell High 35839781
2022 Cryo-EM structures of Giardia Get3 in five states (apo-open, apo-closed, ATP-bound, ADP-bound post-hydrolysis, and with TA client) showed that after ATP hydrolysis Get3 reorganizes the client-binding domain (CBD) to accommodate and shield the client transmembrane helix; the ATP-bound state stabilizes an occluded CBD configuration, resolving how nucleotide drives conformational transitions across the full targeting cycle. Cryo-EM structure determination of Get3 in five nucleotide/conformational states (including Get3-client complex) Nature structural & molecular biology High 35851188
2025 Cryo-EM structure (3.2 Å) of the S. cerevisiae Get3–Get4/5 complex showed that Get4/5 remodels Get3's TA-binding chamber by unfolding helices forming the lateral walls ('lateral gate'), making the chamber more solvent accessible; mutagenesis of lateral gate residues influenced Get3 binding affinity for Get4/5 and its ATPase activity; the Sgt2-binding domain of Get5 is positioned near the lateral gate opening, supporting a model of lateral TA transfer from Sgt2 to Get3. Cryo-EM structure determination + molecular dynamics simulation + site-directed mutagenesis + ATPase activity assay + binding assay The Journal of biological chemistry High 40902977
2014 ASNA1 contains a non-canonical FFAT-like motif that mediates direct interaction with the MSP domain of the ER membrane protein VAPB, physically linking the TRC40/ASNA1 TA-targeting complex to VAPB at the ER membrane. Co-immunoprecipitation + motif identification + direct binding assay BMC biology Medium 24885147
2021 ASNA-1 exists in two redox states that control distinct functions: the reduced state mediates cisplatin resistance and TA protein targeting; an ASNA-1 point mutant preferentially in the oxidized state was sensitive to cisplatin and defective for TA protein targeting but showed normal insulin secretion; cisplatin-induced ROS drives ASNA-1 into the oxidized form and selectively prevents an ASNA-1-dependent TA substrate from reaching the ER. C. elegans genetics with redox-state point mutants + in vivo TA protein targeting assay + cisplatin sensitivity assay + ROS measurement Scientific reports Medium 33883621
2006 Get3 biochemically interacts with the transmembrane domain proteins Get1/Mdm39 and Get2/Rmd7 in yeast; deletion of GET3 suppresses phenotypes of get1 and get2 mutants including sporulation defects; genetic interactions with NPL4 in the ubiquitin-proteasome system implicate Get3 in multiple membrane-dependent pathways. Co-immunoprecipitation + yeast genetic epistasis (suppressor analysis, double mutants) Genetics Medium 16816426
2007 ASNA-1 functions nonautonomously to regulate insulin secretion in C. elegans; expressed in insulin-producing intestinal cells, asna-1 mutants show reduced insulin secretion while overexpression mimics insulin overexpression effects; human ASNA1 is highly expressed in pancreatic beta cells and regulates insulin secretion in cultured cells, demonstrating an evolutionarily conserved role in insulin secretion. C. elegans genetics (asna-1 null and overexpression) + insulin secretion assays in C. elegans + human ASNA1 knockdown/overexpression in cultured beta cells Cell Medium 17289575
2006 Homozygous Asna1 knockout mice die between embryonic day 3.5 and 8.5, demonstrating that Asna1 is essential for early embryonic development in mammals. Homologous recombination knockout mouse generation + embryonic lethality analysis FEBS letters Medium 16797549
2015 Beta-cell-specific inactivation of Asna1 in mice caused hypoinsulinemia, impaired insulin secretion, and rapidly progressive diabetes; Asna1 loss perturbed plasma membrane-to-TGN and Golgi-to-ER retrograde transport and caused ER stress in beta cells; pharmacological inhibition of retrograde transport in isolated islets mimicked the Asna1 loss-of-function phenotype, linking Asna1 function to retrograde vesicular transport and ER homeostasis. Conditional knockout mouse model (beta-cell-specific) + pharmacological retrograde transport inhibition + ER stress markers Diabetes Medium 26438609
2018 Asna1 inactivation in pancreatic multipotent progenitor cells (MPCs) caused redistribution of Golgi TA SNARE proteins syntaxin 5 and syntaxin 6, Golgi fragmentation, integrated stress response activation, and p53-mediated apoptosis leading to pancreatic agenesis; rescue experiments showed the Asna1 ATPase activity and a CXXC di-cysteine motif are required for Golgi integrity and MPC survival; ex vivo inhibition of retrograde transport reproduced the Golgi and syntaxin phenotypes. Conditional knockout mouse model + rescue experiments with ATPase-dead and CXXC mutants + pharmacological retrograde transport inhibition ex vivo + p53 modulator experiments Development Medium 29180572
2016 All three HSV1 tail-anchored proteins (pUL34, pUL56, pUS9) specifically bound to Asna1/TRC40 by yeast two-hybrid; TRC40 depletion by RNAi did not affect virion entry, viral gene expression, or secondary envelopment but specifically reduced release of infectious virions to the extracellular medium by more than 10-fold, identifying a role for TRC40 in viral egress. Yeast two-hybrid protein interaction + siRNA depletion + viral replication and release assays Virology journal Medium 27765046
2026 GET3 (TRC40/ASNA1) directly interacts with the anti-apoptotic protein MCL1 via MCL1's C-terminal hydrophobic tail; GET3 depletion reduced MCL1 protein levels while GET3 overexpression increased them; GET3 deficiency enhanced apoptosis and reduced clonogenic survival, particularly in HeLa cells, and accelerated MCL1 downregulation and apoptosis during prolonged mitotic arrest, identifying MCL1 as a TA-containing cargo of GET3. Degron-mediated GET3 depletion + co-immunoprecipitation of GET3–MCL1 + apoptosis assays + clonogenic survival assay + GET3 overexpression Cell death and differentiation Medium 42098326
2025 Constitutive cardiomyocyte-specific Asna1 knockout caused ventricular myocardial thinning by E16.5 and perinatal lethality; inducible adult cardiomyocyte-specific deletion caused rapid ventricular dilation and early mortality; ASNA1 deficiency destabilized the pre-targeting complex and reduced expression of multiple TA protein substrates, impairing membrane trafficking and vesicular transport; transcriptomic analysis revealed compensatory upregulation of Golgi-to-ER transport genes. Constitutive and inducible cardiomyocyte-specific conditional knockout mouse models + TA protein substrate expression analysis + transcriptomics PLoS genetics Medium 41370295
2025 Neuron-specific deletion of ASNA1 in mice (using SLICK-H-Cre or synapsin-Cre) phenocopied CAML neuron-specific deletion, causing loss of motor neuron cell bodies, hind limb weakness, and paralysis; identifying a cell-autonomous role for the ASNA1/TRC40 TA protein insertion machinery in motor neuron survival. Neuron-specific conditional knockout mouse (synapsin-Cre and SLICK-H-Cre) + spinal cord histology PLoS genetics Medium 39823474
2024 Oxidized TRC40 (human Get3/ASNA1) forms chaperone-active tetramers and high-molecular-weight complexes that prevent aggregation of unfolding proteins; acute oxidative stress causes reversible formation of distinct TRC40 foci co-localizing with Hsp70 and Hsp110; TRC40 is essential for cell survival under ATP-depleting oxidative stress conditions, counteracting accumulation of mis- and unfolded proteins. Biochemical chaperone activity assay + native PAGE oligomer analysis + live-cell fluorescence imaging of TRC40 foci + genetic depletion under oxidative stress bioRxivpreprint Medium

Source papers

Stage 0 corpus · 53 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2009 The structural basis of tail-anchored membrane protein recognition by Get3. Nature 155 19675567
2015 Protein targeting. Structure of the Get3 targeting factor in complex with its membrane protein cargo. Science (New York, N.Y.) 101 25745174
2011 Structural basis for tail-anchored membrane protein biogenesis by the Get3-receptor complex. Science (New York, N.Y.) 101 21719644
2007 ASNA-1 positively regulates insulin secretion in C. elegans and mammalian cells. Cell 101 17289575
2011 WRB is the receptor for TRC40/Asna1-mediated insertion of tail-anchored proteins into the ER membrane. Journal of cell science 98 21444755
2009 Structural insights into tail-anchored protein binding and membrane insertion by Get3. Proceedings of the National Academy of Sciences of the United States of America 88 19948960
2013 Tail-anchored PEX26 targets peroxisomes via a PEX19-dependent and TRC40-independent class I pathway. The Journal of cell biology 81 23460677
2009 Model for eukaryotic tail-anchored protein binding based on the structure of Get3. Proceedings of the National Academy of Sciences of the United States of America 79 19706470
2010 Crystal structure of Get4-Get5 complex and its interactions with Sgt2, Get3, and Ydj1. The Journal of biological chemistry 77 20106980
2010 Structural characterization of the Get4/Get5 complex and its interaction with Get3. Proceedings of the National Academy of Sciences of the United States of America 71 20554915
2014 The protein targeting factor Get3 functions as ATP-independent chaperone under oxidative stress conditions. Molecular cell 62 25242142
2009 The crystal structures of yeast Get3 suggest a mechanism for tail-anchored protein membrane insertion. PloS one 58 19956640
2012 TRC40 can deliver short secretory proteins to the Sec61 translocon. Journal of cell science 53 22505607
2010 Asna1/TRC40-mediated membrane insertion of tail-anchored proteins. Journal of cell science 52 20375064
2014 Crystal structure of ATP-bound Get3-Get4-Get5 complex reveals regulation of Get3 by Get4. Nature structural & molecular biology 51 24727835
2012 Nucleotide-dependent mechanism of Get3 as elucidated from free energy calculations. Proceedings of the National Academy of Sciences of the United States of America 49 22547793
2009 Structural insight into the membrane insertion of tail-anchored proteins by Get3. Genes to cells : devoted to molecular & cellular mechanisms 49 20015340
2012 Get3 is a holdase chaperone and moves to deposition sites for aggregated proteins when membrane targeting is blocked. Journal of cell science 48 23203805
2006 Targeted disruption of the mouse Asna1 gene results in embryonic lethality. FEBS letters 47 16797549
2006 The conserved ATPase Get3/Arr4 modulates the activity of membrane-associated proteins in Saccharomyces cerevisiae. Genetics 46 16816426
2011 Tail-anchor targeting by a Get3 tetramer: the structure of an archaeal homologue. The EMBO journal 36 22124326
2010 Structures of Get3, Get4, and Get5 provide new models for TA membrane protein targeting. Structure (London, England : 1993) 35 20696390
2014 VAPB/ALS8 interacts with FFAT-like proteins including the p97 cofactor FAF1 and the ASNA1 ATPase. BMC biology 31 24885147
2011 Post-translational membrane insertion of tail-anchored transmembrane EF-hand Ca2+ sensor calneurons requires the TRC40/Asna1 protein chaperone. The Journal of biological chemistry 28 21878631
2015 Emery-Dreifuss muscular dystrophy mutations impair TRC40-mediated targeting of emerin to the inner nuclear membrane. Journal of cell science 27 26675233
2016 Tail-anchored Protein Insertion in Mammals: FUNCTION AND RECIPROCAL INTERACTIONS OF THE TWO SUBUNITS OF THE TRC40 RECEPTOR. The Journal of biological chemistry 26 27226539
2015 Asna1/TRC40 Controls β-Cell Function and Endoplasmic Reticulum Homeostasis by Ensuring Retrograde Transport. Diabetes 26 26438609
2012 Get1 stabilizes an open dimer conformation of get3 ATPase by binding two distinct interfaces. Journal of molecular biology 24 22684149
2010 ASNA-1 activity modulates sensitivity to cisplatin. Cancer research 22 20966125
2019 Retro-2 protects cells from ricin toxicity by inhibiting ASNA1-mediated ER targeting and insertion of tail-anchored proteins. eLife 21 31674906
2008 ASNA1, an ATPase targeting tail-anchored proteins, regulates melanoma cell growth and sensitivity to cisplatin and arsenite. Cancer chemotherapy and pharmacology 20 18478230
2019 The natural history of Get3-like chaperones. Traffic (Copenhagen, Denmark) 19 30972921
2019 A trap mutant reveals the physiological client spectrum of TRC40. Journal of cell science 19 31182645
2019 The WRB Subunit of the Get3 Receptor is Required for the Correct Integration of its Partner CAML into the ER. Scientific reports 17 31417168
2022 Structurally derived universal mechanism for the catalytic cycle of the tail-anchored targeting factor Get3. Nature structural & molecular biology 15 35851188
2011 Interaction surface and topology of Get3-Get4-Get5 protein complex, involved in targeting tail-anchored proteins to endoplasmic reticulum. The Journal of biological chemistry 13 22190685
2009 Increased sensitivity to platinating agents and arsenite in human ovarian cancer by downregulation of ASNA1. Oncology reports 13 19724867
2001 Genomic organization and chromosomal localization of the Asna1 gene, a mouse homologue of a bacterial arsenic-translocating ATPase gene. Gene 13 11470536
2021 Molecular characterization and analysis of the ATPase ASNA1 homolog gene of Eimeria tenella in a drug sensitive strain and drug resistant strains. International journal for parasitology. Drugs and drug resistance 11 33639573
2019 Biallelic Variants in ASNA1, Encoding a Cytosolic Targeting Factor of Tail-Anchored Proteins, Cause Rapidly Progressive Pediatric Cardiomyopathy. Circulation. Genomic and precision medicine 11 31461301
2022 From guide to guard-activation mechanism of the stress-sensing chaperone Get3. Molecular cell 10 35839781
2016 Asna1/TRC40 that mediates membrane insertion of tail-anchored proteins is required for efficient release of Herpes simplex virus 1 virions. Virology journal 8 27765046
2018 The ATPase activity of Asna1/TRC40 is required for pancreatic progenitor cell survival. Development (Cambridge, England) 7 29180572
2021 Alternative redox forms of ASNA-1 separate insulin signaling from tail-anchored protein targeting and cisplatin resistance in C. elegans. Scientific reports 6 33883621
2024 A distinct dimer configuration of a diatom Get3 forming a tetrameric complex with its tail-anchored membrane cargo. BMC biology 3 38867239
2022 Identification of C. elegans ASNA-1 domains and tissue requirements that differentially influence platinum sensitivity and growth control. PLoS genetics 2 36480541
2011 It takes two to Get3. Structure (London, England : 1993) 2 22000508
2009 Preliminary X-ray crystallographic studies of yeast Get3. Acta crystallographica. Section F, Structural biology and crystallization communications 1 19407384
2026 Diversity of GET3 chaperones in tail-anchored protein insertion. Journal of experimental botany 0 41764646
2026 GET3 regulates apoptosis via tail-anchoring of MCL1. Cell death and differentiation 0 42098326
2025 Tail Anchored protein insertion mediated by CAML and TRC40 links to neuromuscular function in mice. PLoS genetics 0 39823474
2025 Get4/5-mediated remodeling of Get3's substrate-binding chamber: Insights into tail-anchored protein targeting by the GET pathway. The Journal of biological chemistry 0 40902977
2025 ASNA1 is essential for cardiac development and function by regulating tail-anchored protein stability and vesicular transport in cardiomyocytes. PLoS genetics 0 41370295

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