Affinage

IST1

IST1 homolog · UniProt P53990

Length
364 aa
Mass
39.8 kDa
Annotated
2026-06-10
24 papers in source corpus 18 papers cited in narrative 18 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

IST1 is a divergent ESCRT-III family member that functions as a bidirectional regulator of the VPS4 AAA-ATPase and an adaptor that recruits MIT-domain effectors to sites of ESCRT-III-mediated membrane remodeling (PMID:18032582, PMID:19477918). Its N-terminal core adopts an ESCRT-III subunit-like fold, while its C-terminus presents two distinct MIT-interacting motifs (MIM1 and MIM2) that engage the MIT domains of partner proteins through structurally discriminable binding modes (PMID:19129479, PMID:19477918, PMID:25657007). IST1 inhibits VPS4 ATPase activity when bound alone—through its MIM and a conserved ELYC surface—but binding of Did2/CHMP1 converts it into a VPS4 stimulator, coupling its conformational state to ESCRT-III disassembly (PMID:26515066). At the midbody, IST1 and CHMP1 co-recruit VPS4 and are required for cytokinetic abscission, and IST1 additionally recruits the protease calpain-7 (CAPN7) and spartin to the abscission site, with CAPN7 proteolytic activity executing abscission and maintaining the NoCut checkpoint (PMID:19129479, PMID:20719964, PMID:37772788). Beyond cytokinesis, IST1 partners with CHMP1B to drive normal-topology scission of endosomal recycling carriers, controlling transferrin and mannose-6-phosphate receptor trafficking, and is recruited to endosomal subdomains alternately by SNX15 and CHMP1B (PMID:37926552, PMID:38635626). IST1 also promotes assembly of a CHMP2B/CHMP4B-containing ESCRT-III complex required for autophagosome-lysosome fusion (PMID:31223056), and engages the Ca2+-sensor ALG-2 via a Met-Pro repeat in a Ca2+-dependent manner (PMID:23649269).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 2007 High

    Established IST1 as a regulator of the VPS4 ATPase with opposing activities, answering how a single factor can both promote and restrain ESCRT machinery turnover.

    Evidence Genetic and co-immunoprecipitation analysis of Ist1 with Did2 and Vps4 in S. cerevisiae

    PMID:18032582 PMID:18032584

    Open questions at the time
    • Structural basis of the dual regulation not yet defined
    • Mechanism distinguishing the recruitment versus inhibitory pools unclear
  2. 2009 High

    Defined IST1 as a divergent ESCRT-III subunit and demonstrated its functional role in human cytokinetic abscission, distinguishing it from ESCRT functions in viral budding.

    Evidence RNAi cytokinesis phenotyping, NMR of MIM1/MIM2-VPS4 MIT interaction, midbody immunofluorescence, and HIV-1 budding assay in HeLa cells; crystal structure of Ist1NTD with Did2

    PMID:19129479 PMID:19477918

    Open questions at the time
    • How IST1-CHMP1 cooperatively recruit VPS4 mechanistically
    • Whether abscission requires VPS4 stimulation versus inhibition
  3. 2010 High

    Identified spartin and calpain-7 as IST1-specific MIT-domain effectors, expanding the abscission interactome and revealing IST1 can activate an associated protease.

    Evidence Yeast two-hybrid, SPR, structure-based mutagenesis with dominant-negative rescue for spartin; GST pulldown and in vitro autolysis assay with catalytic-dead mutant for calpain-7

    PMID:20719964 PMID:20849418

    Open questions at the time
    • Physiological substrates of calpain-7 at the midbody not identified
    • Calpain-7 activation finding is single-lab in vitro
  4. 2013 Medium

    Showed IST1 engages the Ca2+-sensor ALG-2 through a distinctive Met-Pro repeat in a Ca2+-dependent, CHMP1-enhanced manner, linking ESCRT-III to calcium signaling.

    Evidence Far-Western blotting and pulldown with deletion mutants of IST1 and GST-ALG-2

    PMID:23649269

    Open questions at the time
    • Cellular consequence of the IST1-ALG-2 interaction not established
    • Single-lab binding study without functional readout
  5. 2015 High

    Resolved the structural logic of IST1 MIM selectivity and the conformational switch governing VPS4, explaining how partner identity dictates IST1 binding mode and VPS4 outcome.

    Evidence Crystal structures of IST1 MIM with VPS4, LIP5, and Spartin MIT domains; ATPase and in vitro ESCRT-III disassembly assays with conformational mutants

    PMID:25657007 PMID:26515066

    Open questions at the time
    • How the conformational switch is triggered in cells
    • Whether ELYC/MIM regulation operates at all IST1 sites of action
  6. 2019 Medium

    Extended IST1 function to autophagosome-lysosome fusion by demonstrating it nucleates a CHMP2B/CHMP4B ESCRT-III complex, with a transcriptional link to tau pathology.

    Evidence Co-IP, AAV-mediated knockdown/overexpression in transgenic mice, autophagy flux assays

    PMID:31223056

    Open questions at the time
    • Direct membrane fusion mechanism not reconstituted
    • Single-lab study
  7. 2022 Medium

    Revealed that ubiquitination of IST1 controls its endosomal recruitment and cargo recycling, adding a post-translational layer to IST1 regulation.

    Evidence Ubiquitination assays, genetic analysis of Cdc48/Npl4, and cargo recycling microscopy in S. cerevisiae

    PMID:36125415

    Open questions at the time
    • Ubiquitin ligase for IST1 not identified
    • Whether mammalian IST1 is similarly regulated unknown
  8. 2023 High

    Mapped CAPN7's tandem MIT engagement of both IST1 MIM motifs and proved this interaction plus CAPN7 catalytic activity are required for abscission and NoCut checkpoint, and defined IST1-CHMP1B in endosomal carrier scission with SNX15 as an alternate recruiter.

    Evidence Crystallography of CAPN7 MIT-IST1 MIM complexes with mutagenesis and depletion/rescue abscission/checkpoint assays; siRNA cargo tracking and SNX15 co-IP

    PMID:37772788 PMID:37926552

    Open questions at the time
    • CAPN7 substrates executing abscission still unidentified
    • How SNX15 versus CHMP1B recruitment is switched in vivo
  9. 2024 Medium

    A selective chemical disruptor of the IST1-CHMP1B interface demonstrated that this copolymer is specifically required for endosomal recycling but dispensable for cytokinesis and MVB sorting, separating IST1's functional modes.

    Evidence Pseudonatural product inhibitor with transferrin recycling, LC3 lipidation, and MVB/EV biogenesis assays

    PMID:38635626

    Open questions at the time
    • Mechanism linking stalled endosomes to noncanonical LC3 lipidation unresolved
    • Single-lab chemical-tool study
  10. 2025 Medium

    Endogenous imaging and effector studies defined distinct IST1 endosomal pools with growth-factor-responsive dynamics and established IST1 as the mediator of Spastin-dependent AMPAR recycling in neurons.

    Evidence CRISPR endogenous tagging with high-speed live imaging; co-IP, siRNA rescue, and mEPSC electrophysiology for the Spastin-IST1 axis

    PMID:41060239 PMID:41296224

    Open questions at the time
    • Molecular determinants distinguishing the two endosomal pools unknown
    • Spastin-IST1 neuronal axis is single-lab

Open questions

Synthesis pass · forward-looking unresolved questions
  • How IST1's conformational/regulatory state, ubiquitination, and partner selection are coordinated to direct it among its distinct membrane-remodeling tasks (abscission, endosomal recycling, autophagy) in cells remains unresolved.
  • No unified model for how IST1 is partitioned among its functional sites
  • Trigger of the inhibitor-to-stimulator switch in vivo unknown
  • Substrate repertoire of IST1-recruited CAPN7 undefined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 3 GO:0005198 structural molecule activity 2 GO:0098772 molecular function regulator activity 2
Localization
GO:0005768 endosome 3 GO:0005856 cytoskeleton 2
Pathway
R-HSA-1640170 Cell Cycle 2 R-HSA-5653656 Vesicle-mediated transport 2 R-HSA-1852241 Organelle biogenesis and maintenance 1 R-HSA-9612973 Autophagy 1
Complex memberships
ESCRT-IIIIST1-CHMP1B copolymer

Evidence

Reading pass · 18 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2007 Yeast Ist1 has a dual role in regulating Vps4: it localizes to the ESCRT machinery via Did2 where it positively recruits Vps4, and it also negatively regulates Vps4 by forming an Ist1-Vps4 heterodimer in which Vps4 cannot bind the ESCRT machinery. Genetic and biochemical analysis in S. cerevisiae, including co-immunoprecipitation and mutant analysis Molecular biology of the cell High 18032582
2007 Yeast Ist1 forms a specific physical complex with Did2, and both Ist1-Did2 and Vta1-Vps60 compose two independent functional units that modulate late steps in MVB sorting; Ist1 endosomal recruitment depends on ESCRT-III. Synthetic genetic analysis (double mutants), co-immunoprecipitation, endosomal localization assays in S. cerevisiae Molecular biology of the cell High 18032584
2009 Human IST1 is required for efficient cytokinetic abscission in HeLa cells; IST1 contains two distinct MIT-interacting motifs (MIM1 and MIM2) at its C-terminus that bind different grooves of the VPS4 MIT domain; IST1 and CHMP1 co-recruit VPS4 to the midbody, and depletion of either blocks VPS4 recruitment and abscission. IST1 depletion does not inhibit HIV-1 budding. RNAi knockdown with cytokinesis phenotype readout, NMR spectroscopy, mutagenesis, midbody localization by immunofluorescence, viral budding assay Molecular biology of the cell High 19129479
2009 The crystal structure of the Ist1 N-terminal domain reveals an ESCRT-III subunit-like fold, identifying Ist1 as a divergent ESCRT-III family member; Ist1NTD specifically binds the Did2 C-terminal MIM1 via a novel MIM-binding structural motif, revealing a mechanism for intermolecular ESCRT-III subunit association. X-ray crystallography of Ist1NTD alone and co-crystallized with Did2 fragment; binding assays Molecular biology of the cell High 19477918
2010 The SPG20 protein spartin is recruited to the midbody through its MIT domain binding to IST1 (but not other CHMP proteins); Ist1 depletion significantly reduces spartin at midbodies; a structure-based mutation (F24D) in spartin MIT domain blocks spartin-IST1 interaction and prevents midbody localization, acting as dominant-negative to impair cytokinesis. Yeast two-hybrid, surface plasmon resonance, siRNA knockdown, immunofluorescence co-localization, dominant-negative rescue experiments Molecular biology of the cell High 20719964
2010 Human IST1 interacts with the tandem MIT domains of calpain 7 via both MIM1 and MIM2, and the IST1 MIM (GST-MIM fusion) enhances autolytic activity of purified calpain 7 in vitro; autolysis requires the catalytic Cys290 and is abolished by N-ethylmaleimide. GST pulldown, co-immunoprecipitation with deletion/point mutants, in vitro autolysis assay with purified proteins, catalytic-dead mutant (C290S) The FEBS journal Medium 20849418
2011 Calpain-7 binds CHMP1B at its second α-helical region (not the canonical MIM1 region), and forms a ternary complex with IST1; coexpression of CHMP1B and IST1 enhances calpain-7 autolysis in cells; ternary complex formation increases calpain-7 recruitment to membrane/organelle fractions. Co-immunoprecipitation with deletion mutants, in vitro pulldown, cell-based autolysis assay (HEK293T), subcellular fractionation Journal of biochemistry Medium 21616915
2013 Mammalian IST1 possesses a distinctive Met-Pro repeat sequence that is essential for Ca2+-dependent interaction with the EF-hand Ca2+-binding protein ALG-2; this interaction is enhanced by co-expression with CHMP1 proteins; IST1 binds ALG-2 by a different mode than other known ALG-2-interacting proteins. Far-Western blotting with biotinylated ALG-2, pulldown assays with deletion mutants of IST1 and GST-ALG-2 mutants Bioscience, biotechnology, and biochemistry Medium 23649269
2015 Crystal structures of IST1 MIM complexed with MIT domains of VPS4, LIP5, and Spartin reveal two distinct binding mechanisms (MIM1 mode vs. MIM3 mode); two phenylalanine residues in IST1 MIM discriminate MIM1 vs. MIM3 binding; structural features in both MIT and MIM determine specificity. X-ray crystallography of three binary complexes, mutagenesis validating key residues The Journal of biological chemistry High 25657007
2015 Ist1 inhibition of Vps4 ATPase activity involves both the MIM and a conserved ELYC surface; an open conformation of the Ist1 core together with MIM interaction stimulates Vps4; binding of Did2 converts Ist1 from a Vps4 inhibitor to a stimulator, and this shift corresponds to altered ESCRT-III disassembly in vitro. ATPase activity assays, in vitro ESCRT-III disassembly assay, mutagenesis of Ist1 conformational elements The Journal of biological chemistry High 26515066
2019 IST1 facilitates association of CHMP2B and CHMP4B/SNF7-2 to form the ESCRT-III complex required for autophagosome-lysosome fusion; lack of IST1 impedes ESCRT-III complex formation and blocks autophagosome-lysosome fusion; MAPT/tau accumulation suppresses IST1 transcription via the ANP32A-INHAT pathway. Co-immunoprecipitation, AAV-mediated overexpression/knockdown in transgenic mice, autophagy flux assays (LC3-II, p62, autophagosome imaging) Autophagy Medium 31223056
2022 In yeast, Ist1 is ubiquitinated, and this ubiquitination is required for proper endosomal recruitment of Ist1 and for recycling of nutrient transporters from endosomes back to the plasma membrane; the AAA-ATPase Cdc48 and its adaptor Npl4 are required for recycling, potentially through regulation of ubiquitinated Ist1. Ubiquitination assays, genetic mutant analysis, fluorescence microscopy of cargo recycling in S. cerevisiae The Journal of cell biology Medium 36125415
2023 Human IST1, together with its binding partner CHMP1B, contributes to scission of early endosomal carriers; depleting IST1 impairs transferrin receptor delivery from early/sorting endosomes to the endocytic recycling compartment and impairs mannose-6-phosphate receptor export; IST1 interacts with the MIT domain-containing sorting nexin SNX15 on endosomes; SNX15 and CHMP1B alternately recruit IST1 to a clathrin-containing subdomain or the base of endosomal tubules. siRNA depletion, live-cell microscopy, kinetic and spatial cargo tracking, co-immunoprecipitation with SNX15 Traffic (Copenhagen, Denmark) Medium 37926552
2023 CAPN7 (Calpain-7) tandem MIT domains bind simultaneously to two distinct MIM motifs on IST1; structure-guided point mutations in either CAPN7 MIT domain disrupt IST1 binding in vitro and in cells; the CAPN7-IST1 interaction is required for CAPN7 recruitment to midbodies, efficient cytokinetic abscission, and NoCut checkpoint arrest; CAPN7 proteolytic activity is also required for abscission and checkpoint maintenance. X-ray crystallography of CAPN7 MIT-IST1 MIM complexes, mutagenesis, co-immunoprecipitation, siRNA depletion/rescue experiments, abscission and checkpoint assays eLife High 37772788
2024 A pseudonatural product compound specifically disrupts the IST1-CHMP1B interaction, inhibiting IST1-CHMP1B copolymer formation required for normal-topology membrane scission; this inhibition rapidly blocks transferrin receptor recycling, causing accumulation in stalled sorting endosomes; stalled endosomes become decorated by lipidated LC3, linking noncanonical LC3 lipidation to IST1-CHMP1B complex disruption. The compound has no impact on cytokinesis, MVB sorting, or extracellular vesicle biogenesis. Chemical inhibitor treatment, transferrin recycling assay, LC3 lipidation assay, live-cell imaging, biogenesis assays for MVBs and extracellular vesicles Proceedings of the National Academy of Sciences of the United States of America Medium 38635626
2025 Endogenous IST1 exists in at least two distinct pools on endosomes (one transient, one relatively stable); upon growth factor stimulation, the stable pool becomes more mobile and the transient pool accumulates more rapidly; ESCRT-III dynamics are distinct from those of other ESCRT complexes, and an intrinsic amount of time is required for ESCRT-mediated ILV biogenesis. CRISPR tagging of endogenous Ist1, high-speed live-cell imaging, fluorescence microscopy The Journal of cell biology Medium 41060239
2025 DeSUMOylated Spastin (Spastin-K427R) exhibits enhanced binding to IST1 compared to wild-type Spastin; co-overexpression of IST1 with Spastin enhances synaptic transmission and spine maturation; IST1 knockdown reduces GluA1 surface levels and abolishes Spastin's effects on AMPAR recycling, indicating IST1 is a key mediator of Spastin-dependent AMPAR endosomal recycling. Co-immunoprecipitation, siRNA knockdown, overexpression, immunofluorescence, electrophysiology (mEPSC recording) Molecular neurobiology Medium 41296224
2025 In mammalian cytokinetic abscission, CHMP2A knockout causes minimal disruption to IST1 and CHMP2B localization at the abscission site, while CHMP4B, CHMP3, and CHMP1B show progressively severe organization defects, supporting a hierarchical ESCRT-III assembly model in which IST1 is among the least dependent on CHMP2A. CHMP2A knockout, live-cell imaging, structured illumination microscopy (SIM), correlative light-electron microscopy (CLEM) bioRxivpreprint Low bio_10.1101_2025.06.24.661003

Source papers

Stage 0 corpus · 24 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2019 MAPT/Tau accumulation represses autophagy flux by disrupting IST1-regulated ESCRT-III complex formation: a vicious cycle in Alzheimer neurodegeneration. Autophagy 163 31223056
2007 Ist1 regulates Vps4 localization and assembly. Molecular biology of the cell 116 18032582
2009 Biochemical analyses of human IST1 and its function in cytokinesis. Molecular biology of the cell 114 19129479
2007 Novel Ist1-Did2 complex functions at a late step in multivesicular body sorting. Molecular biology of the cell 109 18032584
2009 Structural basis of Ist1 function and Ist1-Did2 interaction in the multivesicular body pathway and cytokinesis. Molecular biology of the cell 78 19477918
2010 SPG20 protein spartin is recruited to midbodies by ESCRT-III protein Ist1 and participates in cytokinesis. Molecular biology of the cell 67 20719964
2016 Role of SKD1 Regulators LIP5 and IST1-LIKE1 in Endosomal Sorting and Plant Development. Plant physiology 60 26983994
1998 Expression of Mel-CAM in implantation site intermediate trophoblastic cell line, IST-1, limits its migration on uterine smooth muscle cells. Journal of cell science 51 9701564
2015 Distinct mechanisms of recognizing endosomal sorting complex required for transport III (ESCRT-III) protein IST1 by different microtubule interacting and trafficking (MIT) domains. The Journal of biological chemistry 34 25657007
2010 Autolytic activity of human calpain 7 is enhanced by ESCRT-III-related protein IST1 through MIT-MIM interaction. The FEBS journal 34 20849418
1999 IST1 insertional inactivation of the resB gene: implications for phenotypic switching in Thiobacillus ferrooxidans. FEMS microbiology letters 22 10386372
2022 Recycling of cell surface membrane proteins from yeast endosomes is regulated by ubiquitinated Ist1. The Journal of cell biology 19 36125415
2015 Conformational Changes in the Endosomal Sorting Complex Required for the Transport III Subunit Ist1 Lead to Distinct Modes of ATPase Vps4 Regulation. The Journal of biological chemistry 17 26515066
2024 A chemical inhibitor of IST1-CHMP1B interaction impairs endosomal recycling and induces noncanonical LC3 lipidation. Proceedings of the National Academy of Sciences of the United States of America 11 38635626
2011 Calpain-7 binds to CHMP1B at its second α-helical region and forms a ternary complex with IST1. Journal of biochemistry 11 21616915
2023 IST1 regulates select recycling pathways. Traffic (Copenhagen, Denmark) 10 37926552
2023 The Calpain-7 protease functions together with the ESCRT-III protein IST1 within the midbody to regulate the timing and completion of abscission. eLife 9 37772788
2013 Mammalian ESCRT-III-related protein IST1 has a distinctive met-pro repeat sequence that is essential for interaction with ALG-2 in the presence of Ca2+. Bioscience, biotechnology, and biochemistry 9 23649269
2024 Reduced circ_lrrc49 in trigeminal ganglion contributes to neuropathic pain in mice by downregulating Ist1 and impairing autophagy. Journal of neurochemistry 5 38348636
2025 DeSUMOylation of Spastin Enhances AMPA Receptor Recycling and Synaptic Plasticity via IST1-Dependent Endosomal Sorting. Molecular neurobiology 1 41296224
2025 Insulin receptor substrate family member IST-1 regulates the development of Caenorhabditis elegans age-1 and aap-1 mutants. microPublication biology 0 40642152
2025 Analysis of native Ist1 dynamics reveals multiple pools of ESCRT-III on endosomes. The Journal of cell biology 0 41060239
2025 IST1 alleviated acrylamide-induced apoptosis and cognitive impairment through regulating autophagy flux blockage in SH-SY5Y cells and C57BL/6J mice. Ecotoxicology and environmental safety 0 41187541
2023 IST1 regulates select endosomal recycling pathways. bioRxiv : the preprint server for biology 0 37577466

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