Affinage

PDCD6IP

Programmed cell death 6-interacting protein · UniProt Q8WUM4

Length
868 aa
Mass
96.0 kDa
Annotated
2026-04-29
100 papers in source corpus 49 papers cited in narrative 49 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ALIX (PDCD6IP) is a multifunctional ESCRT-associated adaptor protein that links cargo recognition to ESCRT-III-dependent membrane fission across topologically equivalent processes including multivesicular body (MVB) biogenesis, exosome secretion, retroviral budding, and cytokinetic abscission. ALIX exists in an autoinhibited closed conformation maintained by intramolecular contacts between its proline-rich domain (PRD) and Bro1 domain; Ca²⁺-bound ALG-2 or binding partners relieve this autoinhibition, exposing the Bro1 domain for CHMP4/ESCRT-III recruitment and the V domain for recognition of YPX(n)L motifs on viral Gag proteins and GPCRs, K63-linked polyubiquitin chains, and syntenin–syndecan complexes (PMID:22547407, PMID:23201121, PMID:17350572, PMID:20929444, PMID:27462417, PMID:24637612). Active ALIX dimerizes via its V domain to bridge CHMP4B filaments, and this dimeric scaffold is required for HIV-1 budding, EGFR MVB sorting, and cytokinetic abscission where ALIX is recruited to the midbody by CEP55 and to the abscission site by syndecan-4–syntenin (PMID:19523902, PMID:17853893, PMID:32321914). Beyond endosomal trafficking, ALIX directly binds F-actin, cortactin, and polarity determinants (Par complex, ZO-1) to organize apical actomyosin–tight junction complexes, and Alix knockout in mice causes epithelial polarity defects and hydrocephalus (PMID:27336173, PMID:16966331).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1999 Medium

    The initial identification of ALIX as a Ca²⁺-dependent ALG-2-interacting protein established it as a cytosolic factor potentially linking calcium signaling to cell death pathways, raising the question of its molecular function.

    Evidence Yeast two-hybrid screen with ALG-2, colocalization, and overexpression protection from cell death

    PMID:9880530

    Open questions at the time
    • No endogenous loss-of-function data
    • Mechanism of cell death protection unknown
    • Identity of physiological downstream targets not established
  2. 2003 High

    Discovery that ALIX links retroviral Gag late domains to ESCRT-III/Tsg101 and directly binds CHMP4b via its Bro1 domain established ALIX as a central adaptor in ESCRT-dependent membrane budding, redefining it from a death-related protein to a membrane trafficking factor.

    Evidence Co-immunoprecipitation, GST pulldown, virus budding assays across HIV-1 and EIAV; fluorescence colocalization with CHMP4b at endosomes

    PMID:12860994 PMID:14505569

    Open questions at the time
    • Structural basis of ALIX–cargo and ALIX–CHMP4 interactions unknown
    • Endogenous cellular cargo not identified
  3. 2004 Medium

    Mapping of the ALG-2 binding site to ALIX's PRD PxY repeats, identification of ALIX as an inhibitor of EGFR internalization via antagonism of Cbl–CIN85, and the finding that Src phosphorylation regulates ALIX localization and interactions collectively revealed ALIX as a regulated, multi-domain scaffold at the interface of receptor trafficking and signaling.

    Evidence Mutagenesis mapping ALG-2 site (residues 794–827); siRNA knockdown affecting EGFR internalization; in vitro Src kinase assays with subcellular fractionation

    PMID:14999017 PMID:15456872 PMID:15557335

    Open questions at the time
    • Whether Src phosphorylation regulates ALIX in physiological signaling contexts not shown
    • Relationship between ALG-2 binding and EGFR trafficking not clarified
  4. 2007 High

    Crystal structures of full-length ALIX domains revealed the Bro1–V architecture and defined the molecular basis of dual cargo recognition (YPX(n)L motifs on V domain, CHMP4 helices on Bro1 concave face), while functional studies established ALIX as essential for cytokinetic abscission through CEP55-mediated midbody recruitment.

    Evidence X-ray crystallography of Bro1 and V domains with viral peptides; mutagenesis validated in budding assays; CEP55 interaction by proteomics and Co-IP; RNAi abscission failure

    PMID:17277784 PMID:17350572 PMID:17853893 PMID:18066081 PMID:18641129

    Open questions at the time
    • How ALIX activation is regulated at the midbody versus endosomes not distinguished
    • Full-length ALIX structure in autoinhibited state not solved
  5. 2008 High

    Atomic-resolution structures of Bro1–CHMP4 and ALG-2–ALIX peptide complexes defined the structural logic of both ESCRT-III recruitment and Ca²⁺-dependent activation, revealing the arginine-switch mechanism in ALG-2 that opens a hydrophobic pocket for ALIX binding.

    Evidence X-ray crystallography of Bro1–CHMP4A/B/C peptide complexes and ALG-2–ALIX(799–814) complex with mutagenesis validation

    PMID:18511562 PMID:18940611

    Open questions at the time
    • Whether ALG-2 binding alone is sufficient to relieve autoinhibition not tested
    • Structure of full-length ALG-2–ALIX complex not determined
  6. 2009 High

    SAXS and mutagenesis demonstrated that ALIX functions as a crescent-shaped dimer that bridges CHMP4B filaments, establishing dimerization as a prerequisite for its membrane-remodeling functions.

    Evidence Small-angle X-ray scattering, dimerization-disrupting mutations impairing HIV-1 budding, in vitro CHMP4B filament bridging

    PMID:19523902

    Open questions at the time
    • Whether dimerization is constitutive or regulated not resolved
    • Structural model of dimeric ALIX on membranes not available
  7. 2010 High

    Two independent studies demonstrated that ALIX is autoinhibited by intramolecular PRD–Bro1 and PRD–V domain contacts that mask both the CHMP4- and cargo-binding surfaces, resolving the question of how ALIX activity is kept in check until needed.

    Evidence Systematic truncation/mutagenesis showing PRD folds back against V domain and Bro1; destabilizing mutations increase membrane association and virus budding

    PMID:20929444 PMID:21715492

    Open questions at the time
    • Kinetics and trigger of autoinhibition relief in cells not established
    • Whether different activators use different mechanisms to open ALIX not tested
  8. 2012 High

    ALIX was shown to mediate ubiquitin-independent MVB sorting of GPCRs (PAR1) via YPX(3)L motif recognition and to selectively bind K63-linked polyubiquitin chains, revealing parallel cargo-recognition mechanisms—motif-dependent and ubiquitin-dependent—that diversify ALIX's client repertoire.

    Evidence PAR1 YPX3L mutagenesis and ALIX RNAi blocking MVB sorting; in vitro K63-polyUb binding with V domain mutagenesis

    PMID:22547407 PMID:23201121

    Open questions at the time
    • Whether K63-Ub and YPXnL binding are mutually exclusive on the V domain not clarified
    • Full scope of ALIX-dependent ubiquitin-independent cargos not catalogued
  9. 2014 High

    Identification of the syndecan–syntenin–ALIX axis as the principal pathway for exosome/ILV biogenesis extended ALIX function beyond viral budding and receptor sorting to a major physiological membrane budding pathway regulated by ARF6/PLD2 and heparanase.

    Evidence RNAi, Co-IP, nanoparticle tracking, and electron microscopy in multiple studies

    PMID:24637612 PMID:25732677

    Open questions at the time
    • How LBPA cooperates with syntenin–ALIX at the membrane not mechanistically resolved
    • Relative contribution of syntenin–ALIX versus ESCRT-0/I/II to total ILV budding unknown
  10. 2015 High

    Ca²⁺-dependent ALG-2 binding was shown to fully relieve ALIX autoinhibition and promote CHMP4-dependent membrane association specifically for EGFR MVB sorting, while ALIX's role in cytokinesis and viral budding was ALG-2-independent, establishing context-specific activation mechanisms.

    Evidence Domain-specific mutagenesis with parallel readouts for EGFR sorting, cytokinesis, and EIAV budding

    PMID:27462417

    Open questions at the time
    • What activates ALIX at the midbody if not ALG-2 remains unclear
    • ATG12–ATG3 as activator (PMID:25686249) not integrated with ALG-2 mechanism
  11. 2016 High

    Alix knockout mice revealed an unexpected role in epithelial polarity: ALIX interacts with F-actin, the Par polarity complex, and ZO-1 to organize apical actomyosin–tight junction complexes, and its loss causes hydrocephalus, demonstrating essential developmental functions beyond endosomal trafficking.

    Evidence Alix knockout mouse phenotype (hydrocephalus, epithelial defects), Co-IP with Par complex and ZO-1, electron microscopy

    PMID:27336173

    Open questions at the time
    • Whether polarity defects are secondary to trafficking defects or reflect a direct structural role for ALIX not resolved
    • Which domain of ALIX mediates Par complex interaction not mapped
  12. 2020 High

    In vitro reconstitution on supported lipid bilayers showed that open/active ALIX independently recruits ESCRT-III to LBPA-containing membranes without requiring upstream ESCRTs, while the syndecan-4–syntenin–ALIX module was identified as the abscission-site ESCRT-III recruitment platform distinct from the midbody CEP55 pathway.

    Evidence Reconstitution on supported bilayers; Flemmingsome proteomics with Co-IP and live imaging of abscission

    PMID:32049272 PMID:32321914

    Open questions at the time
    • Whether LBPA directly binds ALIX or acts indirectly not determined
    • Structural basis of syndecan-4–syntenin–ALIX ternary complex not solved
  13. 2020 High

    NMR studies revealed that the ALIX PRD contains three tandem motifs competing for a single TSG101-UEV site and that the tyrosine-rich PRD tail forms reversible amyloid fibrils dissolved by Src phosphorylation, suggesting a regulated phase-transition mechanism for ALIX PRD function.

    Evidence Heteronuclear NMR relaxation dispersion, Congo Red/ThT amyloid assays, TEM, in vitro Src phosphorylation

    PMID:32917811

    Open questions at the time
    • Whether PRD amyloid formation occurs in cells not demonstrated
    • Functional consequence of amyloid dissolution on trafficking or budding not tested
  14. 2023 Medium

    Discovery that ALIX harbors a SUMO-interaction motif (SIM) that recognizes SUMOylated hnRNPA2B1 to sort circular RNA cargo into extracellular vesicles expanded the repertoire of ALIX cargo-recognition mechanisms beyond YPXnL motifs and ubiquitin.

    Evidence Co-IP, ALIX SIM mutagenesis, SUMO modification assays, EV cargo sorting in tumor models

    PMID:37925421

    Open questions at the time
    • SIM-dependent sorting not validated with purified components
    • Generality of SUMO-dependent ALIX cargo recognition beyond this circRNA not explored
    • Single study, awaits independent confirmation

Open questions

Synthesis pass · forward-looking unresolved questions
  • A full-length structure of autoinhibited ALIX, the precise mechanism by which distinct activators (ALG-2, ATG12–ATG3, binding partners) differentially open ALIX for context-specific functions, and the in vivo relevance of PRD amyloid formation remain unresolved.
  • No full-length ALIX structure in closed or open state
  • Relative contribution of ALIX to total cellular ILV/exosome budding not quantified
  • How epithelial polarity and actin-organizing roles relate mechanistically to ESCRT-scaffolding function not integrated

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0060090 molecular adaptor activity 7 GO:0008092 cytoskeletal protein binding 3 GO:0005198 structural molecule activity 2
Localization
GO:0005576 extracellular region 3 GO:0005768 endosome 3 GO:0005856 cytoskeleton 3 GO:0005829 cytosol 2 GO:0031410 cytoplasmic vesicle 2
Pathway
R-HSA-5653656 Vesicle-mediated transport 6 R-HSA-162582 Signal Transduction 4 R-HSA-1640170 Cell Cycle 4 R-HSA-1643685 Disease 4 R-HSA-5357801 Programmed Cell Death 2
Partners
ACTN1ALG2CEP55CHMP4BCIN85CTTNSDCBPTSG101
Complex memberships
CEP55–ALIX–TSG101 (midbody)ESCRT-III (via CHMP4B)syndecan–syntenin–ALIX

Evidence

Reading pass · 49 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 ALIX/AIP1 (PDCD6IP) was identified as a novel protein that interacts with the calcium-binding protein ALG-2 in a Ca2+-dependent manner; the interaction requires calcium as an indispensable requisite, and both proteins colocalize in the cytosol. Overexpression of a truncated AIP1 protects cells from trophic factor withdrawal-induced death. Yeast two-hybrid screening, colocalization, overexpression assays The Journal of biological chemistry Medium 9880530
2003 ALIX/AIP1 functions in HIV-1 and EIAV retrovirus budding by binding to a secondary late-domain (L-domain) region in HIV-1 Gag p6 and to the EIAV p9 L-domain; ALIX also interacts with Tsg101 and ESCRT-III homologs, serving to link viral Gag to the host ESCRT-III budding machinery. Co-immunoprecipitation, binding assays, virus budding assays Cell High 14505569
2003 ALIX interacts directly with CHMP4b (a yeast Snf7 homolog and ESCRT-III component) via its N-terminal Bro1 domain, and this interaction was confirmed by GST pulldown and co-immunoprecipitation; both proteins colocalize at perinuclear endosomal structures and cooperate in multivesicular body (MVB) formation. Yeast two-hybrid, GST pulldown, co-immunoprecipitation, fluorescence microscopy The Journal of biological chemistry High 12860994
2004 The ALG-2-binding site on ALIX maps to amino acid residues 794–827 in the C-terminal proline-rich region, encompassing four PxY repeats; Ca2+-bound ALG-2 interaction with ALIX is required for the punctate subcellular distribution of the ALIX C-terminal fragment. Yeast two-hybrid, co-immunoprecipitation, alanine-scanning mutagenesis, fluorescence microscopy Journal of biochemistry High 14999017
2004 ALIX/AIP1 inhibits EGFR internalization and downregulation by antagonizing the Cbl–CIN85/SETA complex; ALIX reduces Cbl-mediated ubiquitination and the interaction between Cbl and CIN85, while siRNA-mediated knockdown of Alix promotes EGFR internalization and degradation. Co-immunoprecipitation, siRNA knockdown, receptor internalization and ubiquitination assays Molecular and cellular biology Medium 15456872
2004 Alix binds to the transferrin receptor (TfR) cytosolic YTRF motif and contributes to sorting of TfR into exosomes during reticulocyte maturation; AP2 degradation via the proteasome relieves competition at this site and allows hsc70 and Alix to drive TfR exosomal sorting. Co-immunoprecipitation, peptide affinity chromatography, proteasome inhibitor experiments Traffic (Copenhagen, Denmark) Medium 15086793
2004 Paraptosis (a nonapoptotic cell death program induced by the IGF-I receptor) is specifically inhibited by AIP1/Alix, and this inhibition is distinct from apoptosis inhibition; the inhibition does not involve caspase pathways and is mediated via MAP kinases. Overexpression and dominant-negative AIP1, cell death assays, caspase activity assays Cell death and differentiation Medium 15195070
2004 Src kinase phosphorylates ALIX at its C-terminal tyrosine-rich region (via an SH3-SH2 interaction with ALIX), which causes ALIX to translocate from membrane/cytoskeleton to the cytoplasm and disrupts ALIX interactions with CIN85/SETA, EGFR, and Pyk2, antagonizing ALIX's inhibitory role in receptor internalization. In vitro kinase assays, co-immunoprecipitation, subcellular fractionation, overexpression/mutagenesis The Journal of biological chemistry Medium 15557335
2005 Depletion of Alix by RNAi causes redistribution of early endosomes from peripheral to perinuclear location and accumulation of unusual cortactin-containing actin structures, indicating Alix functions in actin-dependent intracellular positioning of endosomes; lysobisphosphatidic acid (LBPA) levels in MVEs are reduced upon Alix depletion. RNAi knockdown, immunofluorescence, immunoelectron microscopy Journal of cell science Medium 15914539
2006 ALIX directly binds filamentous actin (F-actin) through both its N-terminal Bro1 domain and C-terminal proline-rich domain; in cells, ALIX also binds cortactin (via the N-terminal half of the V domain) and α-actinin (via the C-terminal half of the V domain), and is required for lamellipodial cortactin localization and stress fiber assembly. Cell-free actin-binding assays, co-immunoprecipitation, siRNA knockdown, immunofluorescence The Journal of biological chemistry Medium 16966331
2007 Crystal structures of human ALIX reveal it is composed of an N-terminal Bro1 domain and a central V domain (two three-helix bundles forming elongated arms). YPX(n)L late-domain motifs of retroviruses bind a conserved hydrophobic pocket on the second arm of the V domain; CHMP4/ESCRT-III proteins bind a conserved hydrophobic patch on the Bro1 domain; both interactions are required for virus budding. X-ray crystallography, mutagenesis, virus budding assays Cell High 17350572
2007 The structure of the ALIX V domain bound to HIV-1 p6 LYPXnL motif was solved; Phe676 in a large hydrophobic pocket is crucial for binding; overexpression of the V domain alone inhibits HIV-1 release, and this is reversed by mutations blocking V domain–p6 interaction. X-ray crystallography, mutagenesis, virus budding inhibition assay Nature structural & molecular biology High 17277784
2007 Crystal structures of ALIX V domain in complex with YPX(n)L late domains from HIV-1 and EIAV show that distinct late domains bind the same hydrophobic site on the V domain adopting different conformations; binding affinities are context-dependent and key interface residues were validated functionally. X-ray crystallography, surface plasmon resonance, mutagenesis, virus budding assays Nature structural & molecular biology High 18066081
2007 ALIX is recruited to the midbody of dividing cells through direct binding between the central CEP55 'hinge' region and a GPP-based motif in ALIX; depletion of ALIX (and TSG101) inhibits the abscission step of cytokinesis; ALIX point mutants blocking CEP55 and CHMP4/ESCRT-III binding also block abscission. Proteomics, co-immunoprecipitation, RNAi knockdown, dominant-negative mutants, live cell imaging The EMBO journal High 17853893
2007 ALIX potently rescues HIV-1 L-domain mutant budding; this rescue depends on intact ALIX–p6 binding site and the ESCRT-III (CHMP4B)-binding surface on the Bro1 domain; ALIX C-terminal proline-rich domain (PRD) is also required but independently of TSG101/endophilin/CIN85 binding sites. Mutagenesis, virus budding assays, co-immunoprecipitation Journal of virology High 17428861
2007 ALIX is recruited to the cytokinetic midbody via Cep55 interaction through an evolutionarily conserved peptide; disruption of Cep55/Alix/ESCRT-III interactions causes aberrant midbodies and cytokinetic failure; ALIX C-terminus encodes a multimerization activity essential for Alix-dependent HIV-1 release and Tsg101 interaction. Co-immunoprecipitation, dominant-negative mutants, siRNA knockdown, cytokinesis failure assay Proceedings of the National Academy of Sciences of the United States of America High 18641129
2007 HIV-1 ALIX also engages the nucleocapsid (NC) domain of Gag via its Bro1 domain (zinc finger-dependent, RNA-independent) allowing ALIX incorporation into virions independently of the p6 L-domain; NC zinc finger mutations reduce particle production and show functional relationship with the p6 PTAP motif. Co-immunoprecipitation, nuclease treatment, HIV budding assays, ALIX overexpression rescue Journal of virology Medium 18032513
2008 Crystal structures of the ALIX Bro1 domain bound to C-terminal peptides of CHMP4A-C reveal that CHMP4 C-terminal amphipathic helices bind across the concave surface of Bro1; key hydrophobic residues (M/L/IxxLxxW pattern) mediate binding; mutations in exposed Bro1 surface residues block ALIX-dependent HIV-1 budding. X-ray crystallography, mutagenesis, HIV-1 budding assays Proceedings of the National Academy of Sciences of the United States of America High 18511562
2008 ALG-2 crystal structure in complex with the Alix 799-814 peptide reveals a Ca2+/EF3-driven arginine switch: Ca2+ binding to EF3 moves Arg125 to open a hydrophobic pocket that accommodates the PPYP motif of Alix, explaining the Ca2+-dependence of ALG-2/Alix association. X-ray crystallography, in vitro binding assays with mutant proteins Structure (London, England : 1993) High 18940611
2008 AIP1 (DAB2IP) interacts with IRE1 via its PH domain and facilitates IRE1 dimerization, a critical step for IRE1 signaling; AIP1-deficient cells and mice show selectively impaired ER stress-induced IRE1-JNK/XBP-1 signaling but normal PERK-CHOP signaling. AIP1 knockout mice, co-immunoprecipitation, reconstitution with deletion mutants, in vivo signaling assays The Journal of biological chemistry High 18281285
2009 ALIX dimerizes via its V domain in solution; SAXS modeling reveals a crescent-shaped dimeric conformation; dimeric ALIX colocalizes with and bridges CHMP4B filaments; dimerization-disrupting mutations impair HIV-1 budding, indicating dimeric ALIX is the active form that interacts with ESCRT-III CHMP4 polymers. SAXS, mutagenesis, co-localization, HIV-1 budding assay, in vitro CHMP4B filament bridging Structure (London, England : 1993) High 19523902
2009 ALG-2 acts as a Ca2+-dependent adaptor that bridges Alix and TSG101; the ALG-2-binding site on Alix (not the PSAP/TSG101-UEV binding site) is required for Ca2+-dependent TSG101 pulldown; ALG-2 dimerization and Ca2+ binding are both required for bridging. Strep-pulldown with mutants, ALG-2 knockdown, recombinant protein reconstitution Biochemical and biophysical research communications Medium 19520058
2010 The ALIX C-terminal proline-rich region (PRR) folds back against upstream domains and auto-inhibits V domain binding to viral late domains; mutations that destabilize this closed conformation open the V domain, increase ALIX membrane association, and enhance virus budding. Mutagenesis, in vitro binding assays, SAXS/structural modeling, virus budding assays Journal of virology High 21715492
2010 An intramolecular interaction between Patch 2 in the Bro1 domain and the TSG101-docking site in the ALIX PRD locks ALIX in a closed, autoinhibited conformation that prevents CHMP4 and retroviral Gag binding; relieving this intramolecular interaction (by expressing binding partners or deleting interaction sites) enables ALIX to interact with CHMP4 and facilitates membrane association and EIAV budding. ALIX truncation and mutagenesis, co-immunoprecipitation, EIAV budding assay The Biochemical journal High 20929444
2010 SIVmac239 and SIVagmTan-1 possess divergent ALIX-binding late domains distinct from canonical YPX(n)L; crystal structures reveal that anchoring tyrosines and adjacent hydrophobic residues contact the same ALIX V domain hydrophobic pocket used by HIV-1/EIAV late domains. X-ray crystallography, late-domain mapping, virus budding assays Journal of virology High 20962096
2011 Alix Bro1 domain Surface 1 (concave face, CHMP4 binding site), Surface 2 (narrow end), and a unique extended loop exposing Phe105 all independently contribute to ALIX's ability to stimulate HIV-1 budding, as established by mutagenesis guided by the BROX crystal structure. X-ray crystallography (BROX), ALIX mutagenesis, HIV-1 budding assays PloS one Medium 22162750
2012 ALIX binds the V domain to a YPX(3)L motif in the GPCR PAR1 and mediates ubiquitin-independent ESCRT-III-dependent MVB/lysosomal sorting of PAR1, providing an alternative MVB sorting pathway that bypasses ubiquitin-binding ESCRTs. Co-immunoprecipitation, RNAi knockdown, mutagenesis of YPX3L motif, MVB sorting assays The Journal of cell biology High 22547407
2012 ALIX is a Lys63-specific polyubiquitin binding protein; the V domain directly and selectively binds K63-linked polyUb chains (preference for chains ≥3 Ub) via two putative Ub-binding sites on a single α-helical surface; mutations in these sites impair polyUb binding and lentivirus budding. Affinity capture/mass spectrometry, in vitro binding assays, mutagenesis, virus budding assays Developmental cell High 23201121
2012 Alix is ubiquitinated by the Ozz-E3 ligase in skeletal muscle; interaction with Ozz changes Alix conformation and promotes its ubiquitination, regulating Alix levels at cortactin-positive actin remodeling subcompartments; Alix knockdown reduces F-actin levels and impairs cell motility in myotubes. Co-immunoprecipitation, ubiquitination assays, Ozz-/- mouse model, siRNA knockdown The Journal of biological chemistry Medium 22334701
2012 AP-3 adaptor complex binds a tyrosine-based motif in the PAR1 cytoplasmic tail and functions upstream of ALIX to facilitate PAR1 lysosomal sorting; AP-3 promotes PAR1–ALIX interaction, demonstrating a sequential AP-3→ALIX→ESCRT-III pathway for ubiquitin-independent MVB sorting. Co-immunoprecipitation, RNAi knockdown, mutagenesis, receptor sorting assays Molecular biology of the cell Medium 22833563
2014 The syndecan–syntenin–ALIX pathway controls exosome biogenesis and intraluminal budding; ALIX interaction with syntenin and LBPA is required for ILV budding into MVBs; ARF6 and PLD2 regulate this pathway by controlling ILV budding. RNAi knockdown, co-immunoprecipitation, nanoparticle tracking, electron microscopy Nature communications High 24637612
2015 The ATG12–ATG3 conjugate interacts with Alix (PDCD6IP) and controls multiple Alix-dependent processes including late endosome distribution, exosome biogenesis, and viral budding; cells lacking ATG12–ATG3 phenocopy Alix loss-of-function in terms of basal autophagic flux and perinuclear late endosome accumulation. Co-immunoprecipitation, KO cell lines, functional assays for autophagy/exosomes/virus budding Nature cell biology High 25686249
2015 Ca2+-dependent ALG-2 interaction with ALIX completely relieves ALIX's intramolecular autoinhibition and promotes CHMP4-dependent ALIX membrane association; EGFR activation increases ALG-2–ALIX interaction and ALIX membrane association; inhibiting ALG-2-mediated ALIX activation blocks MVB sorting of activated EGFR but not cytokinetic abscission or EIAV budding. Co-immunoprecipitation, mutagenesis, EGFR MVB sorting assays, domain-specific inhibition Cell discovery High 27462417
2015 ALIX mediates MVB sorting and silencing (signal attenuation) of activated ubiquitinated EGFR via V domain interaction with ubiquitinated EGFR and Bro1 domain interaction with membrane-bound CHMP4; inhibiting either ALIX dimerization or CHMP4 interaction blocks EGFR MVB sorting and promotes sustained ERK1/2 activation. EGF stimulation, co-immunoprecipitation, siRNA knockdown, signaling assays The Biochemical journal Medium 25510652
2015 Heparanase stimulates the syndecan–syntenin–ALIX exosome pathway by trimming heparan sulfate chains on syndecans, fostering endosomal membrane budding; exosomal secretion of syntenin-1, syndecan, and CD63 depends on the syntenin–ALIX interaction. RNAi knockdown, exosome quantification, co-immunoprecipitation, electron microscopy Cell research Medium 25732677
2015 Nedd4-1 ubiquitin ligase interacts with ALIX (via Co-IP and yeast two-hybrid) and is recruited to facilitate HIV-1 release through the LYPXnL/ALIX pathway; Nedd4-1 catalytic activity ubiquitinates ALIX itself, and RNAi knockdown of Nedd4-1 eliminates ALIX ubiquitination and impairs ALIX-dependent HIV-1 release. Co-immunoprecipitation, yeast two-hybrid, RNAi knockdown, ubiquitination assay, virus budding assays Journal of virology Medium 20519395
2016 ALIX mediates ubiquitin-independent lysosomal sorting of the P2Y1 purinergic GPCR through a YPX3L motif in the receptor's second intracellular loop; ALIX depletion or mutation of the YPX3L motif prevents P2Y1 sorting into MVB lumen and degradation. RNAi knockdown, site-directed mutagenesis, MVB sorting assays PloS one Medium 27301021
2016 Alix is required for clathrin-independent endocytosis (CIE) of fluid phase and specific cargo (cholera toxin) but not for clathrin-mediated endocytosis; Alix acts with endophilin-A to promote CIE, and is required for fast endocytosis and downstream signaling of the IL-2 receptor. Alix knockout cells, endocytosis assays, siRNA knockdown, signaling assays Scientific reports Medium 27244115
2016 Alix interacts with F-actin, the Par complex, and ZO-1 to mediate assembly of the apically restricted actomyosin–tight junction complex; Alix knockout in mice causes structural epithelial defects (asymmetric cell shape, misplaced cilia, blebbing microvilli), lateral ventricle enlargement, and hydrocephalus. Alix knockout mouse model, co-immunoprecipitation, immunofluorescence, electron microscopy Nature communications High 27336173
2016 Alix interacts with Ago2 (Argonaute 2) in extracellular vesicles; Alix knockdown decreases miRNA levels in EVs without affecting EV number, indicating Alix plays a role in miRNA enrichment/packaging during EV biogenesis. Co-immunoprecipitation, Alix siRNA knockdown, RT-qPCR of EV miRNAs International journal of molecular medicine Medium 26935291
2018 ALIX depletion causes defective PD-L1 trafficking through the MVB and reduces exosomal PD-L1 secretion, leading to redistribution of PD-L1 to the cell surface; ALIX also controls EGFR activity; these effects create an immunosuppressive phenotype dependent on EGFR signaling. siRNA screen, RNAi knockdown, PD-L1 trafficking assays, EGFR phosphorylation assays Cell reports Medium 30021161
2018 Alix undergoes S-palmitoylation (S-acylation), a lipid post-translational modification that influences its interaction with tetraspanin CD9 and maintains the structural organization of exosome-like small extracellular vesicle membranes. Biochemical S-palmitoylation assays, co-immunoprecipitation, biophysical vesicle membrane analysis Biochimica et biophysica acta. General subjects Medium 30251702
2019 During necroptosis, phosphorylated MLKL is removed from membranes by ALIX-syntenin-1-mediated exocytosis (as well as flotillin-mediated endocytosis); ALIX and syntenin-1 are found in MLKL membrane immunoprecipitates by protein cross-linking/affinity purification. Protein cross-linking, affinity purification, functional necroptosis assays Science signaling Medium 31138766
2019 In Drosophila, ALIX is recruited to the cytokinetic midbody via its V domain interaction with centralspindlin component Pavarotti (MKLP1), through a LxxLF motif in Pavarotti binding the conserved hydrophobic pocket of the ALIX V domain (the same pocket used by viral YPXnL/LxxLF motifs); ALIX and ESCRT-III Shrub cooperate for abscission in germline stem cells. Co-immunoprecipitation, mutagenesis, Drosophila genetics, live imaging, cytokinesis assays Current biology : CB High 31607533
2020 An active/open form of ALIX efficiently recruits ESCRT-III proteins to endosomes independent of other ESCRTs but requires lysobisphosphatidic acid (LBPA) in vivo; this ALIX–ESCRT-III pathway can be reconstituted on supported lipid bilayers in vitro and promotes tetraspanin sorting to exosomes. In vitro reconstitution on supported bilayers, RNAi knockdown, endosome fractionation, immunofluorescence The Journal of cell biology High 32049272
2020 ALIX–syntenin–syndecan-4 constitutes a plasma membrane-to-ESCRT module at the cytokinetic abscission site; direct interactions among these three proteins are essential for proper recruitment of ESCRT-III machinery at the abscission site (not at the midbody); depletion of any of the three delays abscission. Proteomics (Flemmingsome), co-immunoprecipitation, RNAi knockdown, live imaging of abscission Nature communications High 32321914
2020 ALIX PRD contains three tandem proline-rich motifs that compete for a single binding site on TSG101-UEV, as demonstrated by NMR relaxation dispersion; the C-terminal tyrosine-rich fragment of ALIX-PRD forms reversible amyloid fibrils that are dissolved by Src kinase-mediated hyperphosphorylation. Heteronuclear NMR spectroscopy, relaxation dispersion analysis, dye-binding assays (Congo Red, ThT), transmission electron microscopy, Src kinase in vitro phosphorylation Proceedings of the National Academy of Sciences of the United States of America High 32917811
2020 Alix and Syntenin-1 are essential for proper subcellular localization and efficient EV secretion of amyloid precursor protein (APP) via an ESCRT-independent pathway; knockdown of either protein alters APP localization to ER and endolysosomal compartments, respectively. Inducible gene knockdown, subcellular fractionation, EV isolation, immunofluorescence BMC molecular and cell biology Medium 32731849
2023 SUMOylated hnRNPA2B1 (at K108, driven by UBC9 upregulated by circTLCD4-RWDD3) is recognized by the SUMO-interaction motif (SIM) of ALIX, which activates ALIX to recruit ESCRT-III, facilitating sorting of circTLCD4-RWDD3 into extracellular vesicles; mutation of ALIX SIM or K108 of hnRNPA2B1 blocks EV packaging. Co-immunoprecipitation, mutagenesis of ALIX SIM, SUMO modification assays, EV cargo sorting assays, in vivo tumor models Signal transduction and targeted therapy Medium 37925421

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2003 AIP1/ALIX is a binding partner for HIV-1 p6 and EIAV p9 functioning in virus budding. Cell 689 14505569
2007 Human ESCRT and ALIX proteins interact with proteins of the midbody and function in cytokinesis. The EMBO journal 594 17853893
2014 Syntenin-ALIX exosome biogenesis and budding into multivesicular bodies are controlled by ARF6 and PLD2. Nature communications 451 24637612
2020 ALIX- and ESCRT-III-dependent sorting of tetraspanins to exosomes. The Journal of cell biology 353 32049272
2004 Paraptosis: mediation by MAP kinases and inhibition by AIP-1/Alix. Cell death and differentiation 314 15195070
2007 Structural and biochemical studies of ALIX/AIP1 and its role in retrovirus budding. Cell 272 17350572
2015 Heparanase activates the syndecan-syntenin-ALIX exosome pathway. Cell research 270 25732677
2015 ATG12-ATG3 interacts with Alix to promote basal autophagic flux and late endosome function. Nature cell biology 258 25686249
2013 ALIX and the multivesicular endosome: ALIX in Wonderland. Trends in cell biology 254 24287454
1999 Cloning of AIP1, a novel protein that associates with the apoptosis-linked gene ALG-2 in a Ca2+-dependent reaction. The Journal of biological chemistry 201 9880530
2008 ALIX-CHMP4 interactions in the human ESCRT pathway. Proceedings of the National Academy of Sciences of the United States of America 194 18511562
2003 The ALG-2-interacting protein Alix associates with CHMP4b, a human homologue of yeast Snf7 that is involved in multivesicular body sorting. The Journal of biological chemistry 184 12860994
2008 Differential requirements for Alix and ESCRT-III in cytokinesis and HIV-1 release. Proceedings of the National Academy of Sciences of the United States of America 182 18641129
2004 Degradation of AP2 during reticulocyte maturation enhances binding of hsc70 and Alix to a common site on TFR for sorting into exosomes. Traffic (Copenhagen, Denmark) 166 15086793
2006 The multiple personalities of Alix. Journal of cell science 156 16868030
2003 Coordinated regulation of actin filament turnover by a high-molecular-weight Srv2/CAP complex, cofilin, profilin, and Aip1. Current biology : CB 149 14680631
2008 Actin disassembly by cofilin, coronin, and Aip1 occurs in bursts and is inhibited by barbed-end cappers. The Journal of cell biology 143 18663144
2007 Beyond Tsg101: the role of Alix in 'ESCRTing' HIV-1. Nature reviews. Microbiology 142 17982468
2012 ALIX binds a YPX(3)L motif of the GPCR PAR1 and mediates ubiquitin-independent ESCRT-III/MVB sorting. The Journal of cell biology 138 22547407
2016 Role of Alix in miRNA packaging during extracellular vesicle biogenesis. International journal of molecular medicine 134 26935291
2007 Structural basis for viral late-domain binding to Alix. Nature structural & molecular biology 132 17277784
2007 Structural and functional studies of ALIX interactions with YPX(n)L late domains of HIV-1 and EIAV. Nature structural & molecular biology 128 18066081
2007 Potent rescue of human immunodeficiency virus type 1 late domain mutants by ALIX/AIP1 depends on its CHMP4 binding site. Journal of virology 127 17428861
2003 AIP1 mediates TNF-alpha-induced ASK1 activation by facilitating dissociation of ASK1 from its inhibitor 14-3-3. The Journal of clinical investigation 127 12813029
2009 Herpes simplex virus type 1 production requires a functional ESCRT-III complex but is independent of TSG101 and ALIX expression. Journal of virology 126 19692479
2018 ALIX Regulates Tumor-Mediated Immunosuppression by Controlling EGFR Activity and PD-L1 Presentation. Cell reports 125 30021161
2004 The actin-interacting protein AIP1 is essential for actin organization and plant development. Current biology : CB 124 14738737
2006 Rapid actin monomer-insensitive depolymerization of Listeria actin comet tails by cofilin, coronin, and Aip1. The Journal of cell biology 120 17060499
2007 Caspase-11 regulates cell migration by promoting Aip1-Cofilin-mediated actin depolymerization. Nature cell biology 114 17293856
2009 A crescent-shaped ALIX dimer targets ESCRT-III CHMP4 filaments. Structure (London, England : 1993) 111 19523902
1999 XAIP1: a Xenopus homologue of yeast actin interacting protein 1 (AIP1), which induces disassembly of actin filaments cooperatively with ADF/cofilin family proteins. Journal of cell science 109 10212149
2001 Yeast PalA/AIP1/Alix homolog Rim20p associates with a PEST-like region and is required for its proteolytic cleavage. Journal of bacteriology 107 11698381
2004 Alix/AIP1 antagonizes epidermal growth factor receptor downregulation by the Cbl-SETA/CIN85 complex. Molecular and cellular biology 101 15456872
2008 AIP1 is critical in transducing IRE1-mediated endoplasmic reticulum stress response. The Journal of biological chemistry 98 18281285
2005 Alix regulates cortical actin and the spatial distribution of endosomes. Journal of cell science 98 15914539
2012 Galectin-3 regulates intracellular trafficking of EGFR through Alix and promotes keratinocyte migration. The Journal of investigative dermatology 97 22785133
2007 Mutations in the cofilin partner Aip1/Wdr1 cause autoinflammatory disease and macrothrombocytopenia. Blood 95 17515402
2019 Flotillin-mediated endocytosis and ALIX-syntenin-1-mediated exocytosis protect the cell membrane from damage caused by necroptosis. Science signaling 92 31138766
2007 Human immunodeficiency virus type 1 Gag engages the Bro1 domain of ALIX/AIP1 through the nucleocapsid. Journal of virology 92 18032513
2015 ESCRT-III-Associated Protein ALIX Mediates High-Affinity Phosphate Transporter Trafficking to Maintain Phosphate Homeostasis in Arabidopsis. The Plant cell 83 26342016
2010 The ESCRT-associated protein Alix recruits the ubiquitin ligase Nedd4-1 to facilitate HIV-1 release through the LYPXnL L domain motif. Journal of virology 81 20519395
2009 Functional role of Alix in HIV-1 replication. Virology 77 19596386
2006 Alix, making a link between apoptosis-linked gene-2, the endosomal sorting complexes required for transport, and neuronal death in vivo. The Journal of neuroscience : the official journal of the Society for Neuroscience 77 16407552
2020 The Flemmingsome reveals an ESCRT-to-membrane coupling via ALIX/syntenin/syndecan-4 required for completion of cytokinesis. Nature communications 76 32321914
2008 AIP1 functions as an endogenous inhibitor of VEGFR2-mediated signaling and inflammatory angiogenesis in mice. The Journal of clinical investigation 76 19033661
2007 RIP1-mediated AIP1 phosphorylation at a 14-3-3-binding site is critical for tumor necrosis factor-induced ASK1-JNK/p38 activation. The Journal of biological chemistry 76 17389591
2010 Loss of Aip1 reveals a role in maintaining the actin monomer pool and an in vivo oligomer assembly pathway. The Journal of cell biology 70 20231387
2012 ALIX is a Lys63-specific polyubiquitin binding protein that functions in retrovirus budding. Developmental cell 67 23201121
2014 Galectin-3 promotes HIV-1 budding via association with Alix and Gag p6. Glycobiology 66 24996823
2014 Aip1 destabilizes cofilin-saturated actin filaments by severing and accelerating monomer dissociation from ends. Current biology : CB 66 25448002
2015 WASH and Tsg101/ALIX-dependent diversion of stress-internalized EGFR from the canonical endocytic pathway. Nature communications 65 26066081
2015 Arabidopsis ALIX is required for the endosomal localization of the deubiquitinating enzyme AMSH3. Proceedings of the National Academy of Sciences of the United States of America 64 26324913
2004 The penta-EF-hand protein ALG-2 interacts with a region containing PxY repeats in Alix/AIP1, which is required for the subcellular punctate distribution of the amino-terminal truncation form of Alix/AIP1. Journal of biochemistry 62 14999017
2007 A novel requirement for C. elegans Alix/ALX-1 in RME-1-mediated membrane transport. Current biology : CB 61 17997305
2008 Structural basis for Ca2+ -dependent formation of ALG-2/Alix peptide complex: Ca2+/EF3-driven arginine switch mechanism. Structure (London, England : 1993) 60 18940611
1999 Cooperation of two actin-binding proteins, cofilin and Aip1, in Saccharomyces cerevisiae. Genes to cells : devoted to molecular & cellular mechanisms 60 10231390
2004 Src phosphorylation of Alix/AIP1 modulates its interaction with binding partners and antagonizes its activities. The Journal of biological chemistry 59 15557335
2017 Functions of actin-interacting protein 1 (AIP1)/WD repeat protein 1 (WDR1) in actin filament dynamics and cytoskeletal regulation. Biochemical and biophysical research communications 58 29056508
2007 The flare gene, which encodes the AIP1 protein of Drosophila, functions to regulate F-actin disassembly in pupal epidermal cells. Genetics 58 17565945
2006 Involvement of the conserved adaptor protein Alix in actin cytoskeleton assembly. The Journal of biological chemistry 58 16966331
2019 Arabidopsis ALIX Regulates Stomatal Aperture and Turnover of Abscisic Acid Receptors. The Plant cell 56 31363038
2006 Do Alix and ALG-2 really control endosomes for better or for worse? Biology of the cell 56 16354163
2014 Aip1 promotes actin filament severing by cofilin and regulates constriction of the cytokinetic contractile ring. The Journal of biological chemistry 54 25451933
2006 HD-PTP and Alix share some membrane-traffic related proteins that interact with their Bro1 domains or proline-rich regions. Archives of biochemistry and biophysics 54 17174262
2015 ALIX Rescues Budding of a Double PTAP/PPEY L-Domain Deletion Mutant of Ebola VP40: A Role for ALIX in Ebola Virus Egress. The Journal of infectious diseases 52 25786915
2006 Interactions between Nef and AIP1 proliferate multivesicular bodies and facilitate egress of HIV-1. Retrovirology 52 16764724
2005 Functions of early (AP-2) and late (AIP1/ALIX) endocytic proteins in equine infectious anemia virus budding. The Journal of biological chemistry 52 16215227
2018 Palmitoylation is a post-translational modification of Alix regulating the membrane organization of exosome-like small extracellular vesicles. Biochimica et biophysica acta. General subjects 49 30251702
2016 ALIX Regulates the Ubiquitin-Independent Lysosomal Sorting of the P2Y1 Purinergic Receptor via a YPX3L Motif. PloS one 49 27301021
2012 AP-3 regulates PAR1 ubiquitin-independent MVB/lysosomal sorting via an ALIX-mediated pathway. Molecular biology of the cell 48 22833563
2011 AIP1 prevents graft arteriosclerosis by inhibiting interferon-γ-dependent smooth muscle cell proliferation and intimal expansion. Circulation research 48 21700930
2009 Penta-EF-hand protein ALG-2 functions as a Ca2+-dependent adaptor that bridges Alix and TSG101. Biochemical and biophysical research communications 47 19520058
2005 AIP1/WDR1 supports mitotic cell rounding. Biochemical and biophysical research communications 47 15629458
2015 ALIX and ESCRT-III coordinately control cytokinetic abscission during germline stem cell division in vivo. PLoS genetics 46 25635693
2023 SUMOylation-triggered ALIX activation modulates extracellular vesicles circTLCD4-RWDD3 to promote lymphatic metastasis of non-small cell lung cancer. Signal transduction and targeted therapy 44 37925421
2013 AIP1 suppresses atherosclerosis by limiting hyperlipidemia-induced inflammation and vascular endothelial dysfunction. Arteriosclerosis, thrombosis, and vascular biology 44 23413429
2016 Alix-mediated assembly of the actomyosin-tight junction polarity complex preserves epithelial polarity and epithelial barrier. Nature communications 42 27336173
2011 Activation of the retroviral budding factor ALIX. Journal of virology 42 21715492
2007 Recruitment of Alix/AIP1 to the plasma membrane by Sendai virus C protein facilitates budding of virus-like particles. Virology 42 18028977
2010 Identification and structural characterization of the ALIX-binding late domains of simian immunodeficiency virus SIVmac239 and SIVagmTan-1. Journal of virology 41 20962096
2002 Hp95 promotes anoikis and inhibits tumorigenicity of HeLa cells. Oncogene 41 12360406
2021 Plasma-Derived Exosomal ALIX as a Novel Biomarker for Diagnosis and Classification of Pancreatic Cancer. Frontiers in oncology 40 34026608
2001 Overexpression of Hp95 induces G1 phase arrest in confluent HeLa cells. Differentiation; research in biological diversity 39 11683497
2006 POSH, a scaffold protein for JNK signaling, binds to ALG-2 and ALIX in Drosophila. FEBS letters 38 16698022
2020 Detection of serum/salivary exosomal Alix in patients with oral squamous cell carcinoma. Oral diseases 36 32688445
2009 An order of magnitude faster AIP1-associated actin disruption than nucleation by the Arp2/3 complex in lamellipodia. PloS one 35 19290054
2016 ALG-2 interacting protein-X (Alix) is essential for clathrin-independent endocytosis and signaling. Scientific reports 34 27244115
2015 ALG-2 activates the MVB sorting function of ALIX through relieving its intramolecular interaction. Cell discovery 34 27462417
2010 Decoding the intrinsic mechanism that prohibits ALIX interaction with ESCRT and viral proteins. The Biochemical journal 34 20929444
2020 Alix and Syntenin-1 direct amyloid precursor protein trafficking into extracellular vesicles. BMC molecular and cell biology 33 32731849
2012 Both internalization and AIP1 association are required for tumor necrosis factor receptor 2-mediated JNK signaling. Arteriosclerosis, thrombosis, and vascular biology 33 22743059
2016 Exosomes in colorectal carcinoma formation: ALIX under the magnifying glass. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 31 27150162
2015 Unravelling the pivotal role of Alix in MVB sorting and silencing of the activated EGFR. The Biochemical journal 31 25510652
2019 ALIX increases protein content and protective function of iPSC-derived exosomes. Journal of molecular medicine (Berlin, Germany) 30 30944935
2020 Proline-rich domain of human ALIX contains multiple TSG101-UEV interaction sites and forms phosphorylation-mediated reversible amyloids. Proceedings of the National Academy of Sciences of the United States of America 27 32917811
2019 Centralspindlin Recruits ALIX to the Midbody during Cytokinetic Abscission in Drosophila via a Mechanism Analogous to Virus Budding. Current biology : CB 27 31607533
2014 AIP1 mediates vascular endothelial cell growth factor receptor-3-dependent angiogenic and lymphangiogenic responses. Arteriosclerosis, thrombosis, and vascular biology 27 24407031
2012 Alix protein is substrate of Ozz-E3 ligase and modulates actin remodeling in skeletal muscle. The Journal of biological chemistry 27 22334701
2022 AIP1 suppresses neovascularization by inhibiting the NOX4-induced NLRP3/NLRP6 imbalance in a murine corneal alkali burn model. Cell communication and signaling : CCS 26 35524333
2011 Structure of the Bro1 domain protein BROX and functional analyses of the ALIX Bro1 domain in HIV-1 budding. PloS one 26 22162750