Affinage

CASTOR1

Cytosolic arginine sensor for mTORC1 subunit 1 · UniProt Q8WTX7

Length
329 aa
Mass
36.3 kDa
Annotated
2026-06-09
16 papers in source corpus 11 papers cited in narrative 10 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CASTOR1 is a cytosolic arginine sensor that gates amino-acid signaling to mTORC1 by controlling the GATOR2 complex (PMID:27487210, PMID:40715445, PMID:40470200). In the arginine-free state it forms a homodimer through its ACT1 and ACT3 domains and engages two MIOS WD40 β-propellers of the GATOR2 cage via a negatively charged surface distal to its ligand pocket, holding GATOR2 in an inhibited state and keeping mTORC1 low (PMID:27648300, PMID:40715445, PMID:40470200). Arginine binds at the interface of the ACT2 and ACT4 domains, and rather than driving a large global rearrangement, it orders two loop regions that sterically occlude the MIOS-binding surface, releasing CASTOR1 from GATOR2 and de-repressing GATOR1 GAP activity to activate lysosomal mTORC1 (PMID:28066558, PMID:30503338, PMID:40715445, PMID:40470200). CASTOR1 activity is additionally tuned post-translationally: AKT phosphorylates CASTOR1 at Ser14, increasing its binding to the E3 ligase RNF167, which catalyzes K29-linked polyubiquitination and proteasomal degradation of CASTOR1 to activate mTORC1 independently of arginine (PMID:33594058). CASTOR1 and its homolog CASTOR2 act as a dual-sensitivity system responding to low and high arginine respectively, together fine-tuning mTORC1 output during processes such as C2C12 myogenesis (PMID:41506264). Through this mTORC1-restraining role, CASTOR1 functions as a brake in disease-relevant settings, including KRAS-driven lung tumorigenesis and KSHV miRNA-driven mTORC1 activation (PMID:31305263, PMID:40313924).

Mechanistic history

Synthesis pass · year-by-year structured walk · 9 steps
  1. 2016 High

    Established the structural basis for how CASTOR1 senses arginine and couples that signal to mTORC1, defining it as a ligand-gated repressor of GATOR2.

    Evidence X-ray crystallography of arginine-bound CASTOR1 with structure-guided mutagenesis and binding assays

    PMID:27487210 PMID:27648300 PMID:28066558

    Open questions at the time
    • Apo-state conformation not resolved in these structures
    • Architecture of the CASTOR1–GATOR2 contact inferred from mutagenesis, not visualized
    • Did not address physiological arginine concentration thresholds
  2. 2016 High

    Defined the domain logic of CASTOR1, separating the homodimerization function (ACT1/ACT3) from the arginine-binding function (ACT2/ACT4).

    Evidence 2.5 Å crystal structure with site-directed mutagenesis of pocket and dimer-interface residues

    PMID:27648300

    Open questions at the time
    • Did not show how loop changes propagate to the GATOR2 interface
    • Functional readouts in cells limited
  3. 2018 High

    Resolved the conformational mechanism debate by showing arginine sensing is a localized loop reordering rather than a global domain swing.

    Evidence Comparison of apo (2.8 Å) and arginine-bound (2.05 Å) CASTOR1 crystal structures

    PMID:30503338

    Open questions at the time
    • How loop ordering translates into loss of GATOR2 binding not demonstrated structurally at the time
    • Single-lab structures
  4. 2019 Medium

    Showed CASTOR1 is a target of pathogen-encoded regulation, linking viral miRNAs to mTORC1 activation through suppression of an arginine sensor.

    Evidence KSHV miRNA 3′-UTR target validation, miRNA knockdown rescue, and mTORC1/colony-formation assays

    PMID:31305263

    Open questions at the time
    • Single lab
    • Relative contribution of miR-K1-5p not firmly quantified
    • In vivo relevance not tested
  5. 2021 High

    Uncovered an arginine-independent control layer in which AKT-driven phosphorylation routes CASTOR1 for ubiquitin-mediated degradation to activate mTORC1.

    Evidence Co-IP, K29-linkage ubiquitination assays, Ser14 phospho-site mutagenesis, RNF167 perturbation, and AKT inhibition

    PMID:33594058

    Open questions at the time
    • Physiological stimuli engaging this axis not fully mapped
    • Crosstalk timing with arginine sensing unresolved
  6. 2022 Medium

    Placed CASTOR1 upstream of mTOR in a cellular phenotype, showing it restrains pro-inflammatory M1 microglial polarization.

    Evidence CASTOR1 overexpression with mTOR-activator (MHY1485) epistasis and M1/M2 marker readouts in primary microglia

    PMID:36454506

    Open questions at the time
    • Single cellular model
    • Endogenous loss-of-function not tested
    • Arginine dependence in this context unexamined
  7. 2025 High

    Captured the intact arginine-free GATOR2–CASTOR1 complex, directly visualizing the MIOS contact and confirming the steric-occlusion switch model.

    Evidence Cryo-EM of the GATOR2–CASTOR1 complex with structural and functional validation

    PMID:40470200 PMID:40715445

    Open questions at the time
    • Dynamics of the binding/release transition not directly observed
    • Stoichiometry relative to full GATOR2 cage occupancy not fully quantified
  8. 2025 Medium

    Provided in vivo evidence that CASTOR1 acts as a tumor suppressor that brakes mTORC1 and interacts with KRAS/ERK signaling.

    Evidence CASTOR1-KO × KRAS G12D mouse model, organoids, phospho-protein readouts, and drug-combination rescue (preprint)

    PMID:40313924

    Open questions at the time
    • Preprint, not peer-reviewed
    • Single lab
    • Direct biochemical basis of AKT/ERK crosstalk not established
  9. 2026 High

    Defined CASTOR1 and CASTOR2 as a dual-sensitivity arginine-sensing pair, extending the single-sensor model to a graded mTORC1 readout with a developmental output.

    Evidence Biochemical binding, conformational analysis, arginine dose-response, and C2C12 knockdown with myogenesis readouts

    PMID:41506264

    Open questions at the time
    • Single lab
    • In vivo myogenesis relevance not tested
    • Quantitative crossover point between the two sensors not fully defined

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the arginine-sensing, AKT/RNF167 degradation, and KRAS/ERK crosstalk inputs are integrated to set mTORC1 output in specific tissues remains unresolved.
  • No unified model linking post-translational degradation with structural ligand sensing in vivo
  • Tissue-specific physiological roles beyond muscle, microglia, and lung unmapped

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140299 molecular sensor activity 6 GO:0098772 molecular function regulator activity 2
Localization
GO:0005829 cytosol 2
Pathway
R-HSA-162582 Signal Transduction 3

Evidence

Reading pass · 10 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2016 Crystal structure of arginine-bound CASTOR1 at 1.8 Å resolution reveals that homodimeric CASTOR1 binds arginine at the interface of two ACT domains, enabling allosteric control of the adjacent GATOR2-binding site; arginine binding triggers dissociation from GATOR2, thereby activating mTORC1 downstream. X-ray crystallography (1.8 Å), structure-guided mutagenesis, biochemical binding assays Nature High 27487210
2016 Crystal structure of human CASTOR1–arginine complex identifies an arginine-binding pocket between the NTD and CTD (ACT) domains; a surface patch on CASTOR1-NTD opposite this pocket mediates direct physical interaction with GATOR2 subunit Mios; mutation of key pocket residues abolishes or diminishes arginine binding, and mutation of the surface patch disrupts CASTOR1 recognition and inhibition of GATOR2. X-ray crystallography, in vitro pull-down assay, site-directed mutagenesis, normal mode analysis Cell discovery High 28066558
2016 Crystal structure of CASTOR1–arginine complex at 2.5 Å shows four tandem ACT domains; ACT1 and ACT3 mediate homodimerization via conserved helix residues, while ACT2 and ACT4 form the arginine-binding pocket; mutagenesis of key binding-pocket residues validates their functional roles in arginine-dependent mTORC1 activation. X-ray crystallography (2.5 Å), site-directed mutagenesis, biochemical assays Cell discovery High 27648300
2018 Comparison of arginine-bound (2.05 Å) and ligand-free (2.8 Å) CASTOR1 crystal structures reveals near-identical conformations except in two loop regions, indicating that CASTOR1 does not undergo large conformational change upon arginine binding; arginine sensing is thus interpreted as a subtle loop-based mechanism rather than a global domain rearrangement. X-ray crystallography (apo and holo structures), structural comparison Biochemical and biophysical research communications High 30503338
2025 Cryo-EM structure of the GATOR2–CASTOR1 complex (arginine-free) shows that two MIOS WD40 β-propellers of the GATOR2 cage engage both subunits of a single CASTOR1 homodimer at a negatively charged interface distal to the arginine pocket; arginine-triggered loop ordering in CASTOR1 sterically blocks this MIOS-binding interface, switching off GATOR2 binding and thereby de-repressing GATOR1 and activating mTORC1. Cryo-electron microscopy, structural analysis, functional validation Nature structural & molecular biology High 40470200 40715445
2021 E3 ubiquitin ligase RNF167 catalyzes K29-linked polyubiquitination and proteasomal degradation of CASTOR1; AKT phosphorylates CASTOR1 at Ser14, which markedly increases CASTOR1 binding to RNF167 (promoting its ubiquitination and degradation) while simultaneously decreasing CASTOR1 affinity for MIOS, resulting in mTORC1 activation independent of arginine. Co-immunoprecipitation, ubiquitination assays, phosphorylation site mutagenesis, RNF167 knockdown/overexpression, AKT inhibitor treatment Nature communications High 33594058
2019 KSHV-encoded miRNAs miR-K4-5p (and likely miR-K1-5p) directly target the CASTOR1 3′-UTR to suppress CASTOR1 expression; knockdown of these miRNAs restores CASTOR1 protein levels and attenuates mTORC1 activation, establishing a viral miRNA–CASTOR1–GATOR2–mTORC1 axis. miRNA target validation (direct targeting of CASTOR1 3′-UTR), miRNA knockdown, CASTOR1/CASTOR2 overexpression, mTORC1 activity assays, soft-agar colony formation The Journal of clinical investigation Medium 31305263
2026 CASTOR1 and CASTOR2 both bind arginine and interact with GATOR2 component Mios; arginine binding induces conformational changes at the ACT2–ACT4 interface causing dissociation from Mios; CASTOR1 responds to low arginine concentrations and CASTOR2 to high arginine concentrations, enabling dual-sensor fine-tuning of mTORC1 activity; in C2C12 muscle cells, CASTOR proteins regulate mTORC1 and myogenesis in an arginine-level-dependent manner. Biochemical binding assays, structural analysis of conformational changes, arginine dose-response experiments, C2C12 cell knockdown with mTORC1 and myogenesis readouts Molecular cell High 41506264
2022 In LPS/IFN-γ-activated microglia, CASTOR1 overexpression inhibits M1 polarization by suppressing mTOR signaling; mTOR activator MHY1485 rescues M1 polarization in CASTOR1-overexpressing cells, placing CASTOR1 upstream of mTOR in microglial polarization. CASTOR1 overexpression, mTOR activator (MHY1485) epistasis, M1/M2 marker expression assays in primary microglia Metabolic brain disease Medium 36454506
2025 CASTOR1 genetic ablation in a KRAS-driven GEMM (LSL-KRAS G12D) enhances lung tumor incidence and accelerates progression; mechanistically, CASTOR1 loss amplifies mTORC1 signaling (elevated p-4EBP1, p-S6) and augments AKT and ERK activation, revealing crosstalk between PI3K/AKT/mTORC1 and KRAS/ERK pathways; mTORC1 and PI3K inhibitors sensitize CASTOR1-deficient resistant tumors to KRAS G12D-targeted therapy. Genetically engineered mouse model (CASTOR1 KO × KRAS G12D), tumor-derived organoids, phospho-protein assays, drug combination rescue bioRxivpreprint Medium 40313924

Source papers

Stage 0 corpus · 16 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2016 Mechanism of arginine sensing by CASTOR1 upstream of mTORC1. Nature 258 27487210
2016 Structural mechanism for the arginine sensing and regulation of CASTOR1 in the mTORC1 signaling pathway. Cell discovery 49 28066558
2021 RNF167 activates mTORC1 and promotes tumorigenesis by targeting CASTOR1 for ubiquitination and degradation. Nature communications 41 33594058
2019 Kaposi sarcoma-associated herpesvirus miRNAs suppress CASTOR1-mediated mTORC1 inhibition to promote tumorigenesis. The Journal of clinical investigation 37 31305263
2016 Structural insight into the arginine-binding specificity of CASTOR1 in amino acid-dependent mTORC1 signaling. Cell discovery 34 27648300
2024 L-arginine alleviates heat stress-induced mammary gland injury through modulating CASTOR1-mTORC1 axis mediated mitochondrial homeostasis. The Science of the total environment 16 38552976
2025 piR-16404 drives ferroptotic liver injury via CASTOR1/mTORC1/GPX4 dysregulation in HepG2 cells and mice: a novel toxicity mechanism of N, N-dimethylformamide. Archives of toxicology 13 40982001
2018 Crystal structures of arginine sensor CASTOR1 in arginine-bound and ligand free states. Biochemical and biophysical research communications 13 30503338
2025 Structural basis for mTORC1 regulation by the CASTOR1-GATOR2 complex. Nature structural & molecular biology 9 40715445
2022 Downregulation of CASTOR1 Inhibits Heat-Stress-Induced Apoptosis and Promotes Casein and Lipid Synthesis in Mammary Epithelial Cells. Journal of agricultural and food chemistry 7 35442666
2022 Castor1 overexpression regulates microglia M1/M2 polarization via inhibiting mTOR pathway. Metabolic brain disease 7 36454504
2025 Upregulating mTOR/S6 K Pathway by CASTOR1 Promotes Astrocyte Proliferation and Myelination in Gpam-/--induced mouse model of cerebral palsy. Molecular neurobiology 3 40234290
2025 CASTOR1: A Novel Tumor Suppressor Linking mTORC1 and KRAS Pathways in Tumorigenesis and Resistance to KRAS-Targeted Therapies in Non-Small Cell Lung Cancer. bioRxiv : the preprint server for biology 1 40313924
2026 CASTOR1 and CASTOR2 respond to different arginine levels to regulate mTORC1 activity. Molecular cell 0 41506264
2026 Putative identification of CASTOR1 as one of the targets of ganoderic acid a via thermal proteome profiling and molecular docking. Frontiers in pharmacology 0 42028441
2025 Structural basis for mTORC1 regulation by the CASTOR1-GATOR2 complex. Research square 0 40470200

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