{"gene":"TSG101","run_date":"2026-06-10T10:51:56","timeline":{"discoveries":[{"year":2001,"finding":"The UEV (ubiquitin E2 variant) domain of TSG101 directly binds the PTAP tetrapeptide 'late domain' motif of HIV-1 Gag p6 protein, and also binds ubiquitin. Depletion of TSG101 by siRNA arrests HIV-1 budding at a late stage; reintroduction rescues budding. Dominant-negative VPS4 also arrests HIV-1 budding, placing TSG101 in the vacuolar protein sorting (VPS)/ESCRT pathway required for HIV-1 egress.","method":"siRNA depletion, co-immunoprecipitation, surface plasmon resonance binding assay, dominant-negative VPS4 overexpression, electron microscopy","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal methods (siRNA, binding assays, dominant-negative, EM), replicated across multiple subsequent studies","pmids":["11595185"],"is_preprint":false},{"year":2001,"finding":"TSG101 interacts with the PTAP-containing late domain of HIV-1 Gag and the PTAP-like motif of Ebola VP40 (EbVp40). Recruitment of TSG101 to assembling virions restores budding competence to late-domain-defective HIV-1, establishing that TSG101 recruitment is the essential function of the viral late domain.","method":"Co-immunoprecipitation, dominant-negative TSG101 overexpression, functional rescue assay, confocal microscopy","journal":"Nature medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction assays with functional rescue, independently replicated","pmids":["11726971"],"is_preprint":false},{"year":2001,"finding":"TSG101 binds to the PTAP motif of HIV-1 Gag p6 through its N-terminal UBC-like domain. Mutation of Tyr-110 (in place of the active-site Cys present in canonical E2 enzymes) and surrounding residues unique to TSG101 impairs p6 binding, showing that TSG101 is a catalytically inactive E2 variant whose distinctive features mediate viral protein interaction.","method":"Yeast two-hybrid, in vitro co-immunoprecipitation with purified proteins, in vivo co-immunoprecipitation in COS cells, site-directed mutagenesis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution with purified proteins + mutagenesis + multiple binding assays in one study","pmids":["11427703"],"is_preprint":false},{"year":2000,"finding":"TSG101 (mammalian VPS23) functions in late endosomal trafficking. TSG101 mutant cells show defects in cathepsin D sorting/maturation, mislocalization of mannose-6-phosphate receptor, and recycling of endocytosed EGF receptors back to the cell surface instead of lysosomal delivery, demonstrating TSG101 is required for cargo delivery to late endosomes/MVBs.","method":"Cell fractionation, Western blotting, receptor trafficking assays in TSG101 mutant cells, pulse-chase","journal":"Traffic (Copenhagen, Denmark)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal trafficking assays in TSG101-deficient cells, single lab","pmids":["11208108"],"is_preprint":false},{"year":2002,"finding":"The crystal/NMR structure of the TSG101 UEV domain reveals it resembles canonical E2 ubiquitin-conjugating enzymes but has an additional N-terminal helix, an extended beta-hairpin, and lacks two C-terminal helices. PTAP peptides bind in a hydrophobic cleft exposed by the absence of C-terminal helices; ubiquitin binds at a novel site surrounding the beta-hairpin. These two binding sites are spatially distinct and independent.","method":"NMR structure determination, chemical shift mapping, surface plasmon resonance binding assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — NMR structure with functional binding site mapping using orthogonal biochemical methods","pmids":["12006492"],"is_preprint":false},{"year":2002,"finding":"Solution structure of the TSG101 UEV domain in complex with the HIV-1 p6 PTAP peptide shows that each PTAP residue makes important contacts; the Ala-Pro dipeptide binds in a deep pocket resembling the X-Pro binding pockets of SH3 and WW domains, located above the vestigial active site.","method":"NMR solution structure determination","journal":"Nature structural biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — atomic-resolution NMR structure of the complex","pmids":["12379843"],"is_preprint":false},{"year":2004,"finding":"Crystal structure of the TSG101 UEV domain in complex with ubiquitin at 2.0 Å resolution shows TSG101 UEV contacts the Ile44 surface and an adjacent loop of ubiquitin through a highly solvated interface. Mutations disrupting this interface inhibit MVB sorting, and the ubiquitin-binding site is distinct from the PTAP-binding site, allowing independent binding of both ligands.","method":"X-ray crystallography, site-directed mutagenesis, functional MVB sorting assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with mutagenesis and functional validation","pmids":["15053872"],"is_preprint":false},{"year":2000,"finding":"TSG101 directly interacts with mammalian VPS28 (hVPS28) through the conserved C-terminal portion of TSG101 to form part of a multiprotein complex (~350 kDa). Upon expression of dominant-negative VPS4, a portion of TSG101 and hVPS28 translocate from cytosol to the surface of aberrant endosomal vacuoles.","method":"Co-immunoprecipitation, chemical cross-linking, gel filtration, confocal microscopy with dominant-negative VPS4","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — direct interaction demonstrated by co-IP and cross-linking with domain mapping, functional localization experiment","pmids":["11134028"],"is_preprint":false},{"year":2003,"finding":"TSG101 interacts with HRS (hepatocyte growth factor-regulated tyrosine kinase substrate) via the UEV domain of TSG101 binding to two proline-rich regions of HRS including a PSAP motif. Disruption of this interaction prevents delivery of EGFR to late endosomes, causes accumulation of ubiquitinated EGFR in early endosomes, and inhibits ligand-induced EGFR down-regulation.","method":"Co-immunoprecipitation, deletion/mutagenesis analysis, confocal microscopy, receptor trafficking assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with domain mapping plus functional consequence (receptor trafficking), single lab with multiple orthogonal methods","pmids":["12802020"],"is_preprint":false},{"year":2003,"finding":"HIV-1 Gag mimics the Tsg101-recruiting activity of the endosomal protein Hrs. The Tsg101 UEV domain binds the PSAP motif (residues 348-351) within Hrs, and Hrs residues 222-777 can recruit Tsg101 and rescue budding of Gag particles lacking native late domains, demonstrating that Hrs normally recruits Tsg101 to the endosomal membrane and HIV Gag usurps this activity.","method":"Co-immunoprecipitation, yeast two-hybrid, functional rescue assay, dominant-negative competition","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding assays plus functional rescue, independently supported by parallel paper (PMID 12802020)","pmids":["12900394"],"is_preprint":false},{"year":2001,"finding":"TSG101 participates in an autoregulatory loop with MDM2: the UBC domain of TSG101 interferes with ubiquitination of MDM2, stabilizing MDM2 and thereby down-regulating p53 protein levels. Conversely, elevated MDM2 promotes TSG101 degradation via the 26S proteasome.","method":"Pulse-chase analysis, Western blotting in wild-type and mutant fibroblasts, ubiquitination assays, MDM2/p53 overexpression experiments","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple biochemical methods in one lab establishing the regulatory loop, but no in vitro reconstitution of the direct inhibition of MDM2 ubiquitination","pmids":["11172000"],"is_preprint":false},{"year":2001,"finding":"TSG101 null embryos accumulate p53 protein (without change in p53 mRNA), and introduction of a p53 null mutation into tsg101-/- embryos rescues the gastrulation defect and extends survival, establishing a functional genetic link between TSG101 and the p53 pathway in vivo.","method":"Gene targeting (knockout mice), genetic epistasis (double knockout), Western blotting, in vivo embryo analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis by double-knockout with defined phenotypic rescue, rigorous in vivo model","pmids":["11172041"],"is_preprint":false},{"year":2002,"finding":"Ubiquitylation of MHC class I by the Kaposi's sarcoma herpesvirus K3 protein requires TSG101 for subsequent degradation in the late endocytic pathway. siRNA depletion of TSG101 prevents MHC class I degradation and preserves its cell surface expression in K3-expressing cells.","method":"siRNA knockdown, flow cytometry, Western blotting, confocal microscopy","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean siRNA knockdown with defined functional readout, single lab","pmids":["12006494"],"is_preprint":false},{"year":2004,"finding":"Tal (Tsg101-associated ligase) is an E3 ubiquitin ligase whose RING finger mediates multiple monoubiquitylation of TSG101. Bivalent binding of TSG101 to a tandem PTAP motif and a central region of Tal is required for Tal-mediated TSG101 ubiquitylation. Tal regulates a TSG101-associated complex responsible for sorting cargo into MVB vesicles and plasma membrane budding.","method":"Co-immunoprecipitation, in vitro ubiquitination assay, deletion/mutagenesis analysis, functional assays (EGFR endocytosis, HIV egress)","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus domain mapping and functional validation, single lab","pmids":["15256501"],"is_preprint":false},{"year":2007,"finding":"Mahogunin E3 ubiquitin ligase interacts with the UEV domain of TSG101 via its PSAP motif and catalyzes monoubiquitylation of TSG101 both in vivo and in vitro. Depletion of Mahogunin disrupts endosome-to-lysosome trafficking of EGFR, resulting in prolonged downstream signaling.","method":"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, EGFR trafficking assays","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus interaction mapping and functional knockdown assay, single lab","pmids":["17229889"],"is_preprint":false},{"year":2007,"finding":"The steadiness box (conserved C-terminal sequence) of TSG101 controls its steady-state level. Tal polyubiquitinates lysine residues in the C-terminus of TSG101 not complexed with other ESCRT-I proteins, leading to proteasomal degradation. VPS28 is a limiting factor, and ESCRT-I complex assembly protects TSG101 from Tal-mediated degradation.","method":"Ubiquitination assays, proteasome inhibitor experiments, siRNA knockdown, co-immunoprecipitation, Western blotting","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — in vitro and in vivo ubiquitination assays with domain mutagenesis and functional complementation, single lab","pmids":["18077552"],"is_preprint":false},{"year":2000,"finding":"The TSG101 protein steady-state level is controlled posttranslationally by an evolutionarily conserved C-terminal sequence termed the 'steadiness box.' Overproduction of TSG101 from adventitious constructs results in compensatory down-regulation of endogenous TSG101 protein (not mRNA), through a proteolytic feedback loop involving the steadiness box.","method":"Western blotting, Northern blotting, deletion mapping of TSG101 constructs, pulse-chase in cultured cells","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple complementary experiments mapping the regulatory sequence, functional consequences assessed, single lab","pmids":["10749147"],"is_preprint":false},{"year":1998,"finding":"TSG101 localization is cell cycle-dependent: it localizes to the nucleus and Golgi complex during interphase, and to mitotic spindles and centrosomes during mitosis. TSG101-deficient cells display multiple microtubule organizing centers, aberrant mitotic spindles, aneuploidy, and nuclear anomalies.","method":"Indirect immunofluorescence, cell fractionation, FACS analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization by immunofluorescence across cell cycle stages with functional phenotyping of deficient cells, single lab","pmids":["9465061"],"is_preprint":false},{"year":2006,"finding":"Depletion of TSG101 inhibits EGF-stimulated MVB formation (number of MVBs per unit cytoplasm), causes tubulation of the early endosome vacuolar domains, and potently inhibits EGF degradation; this is distinct from the role of Hrs, which is required for accumulation of internal vesicles within MVBs but not MVB formation itself.","method":"siRNA depletion, electron microscopy, confocal microscopy, EGF degradation assays","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — siRNA with defined ultrastructural and functional readouts comparing TSG101 and Hrs roles, single lab with multiple orthogonal methods","pmids":["16707569"],"is_preprint":false},{"year":2008,"finding":"In a cell-free reconstitution assay, the ESCRT-I subunit Tsg101 acts as a positive regulator of intraluminal vesicle (ILV) formation within late endosomes, while Alix acts as a negative regulator; ILV budding is inhibited by dominant-negative VPS4, confirming ESCRT dependence.","method":"In vitro budding reconstitution assay, dominant-negative mutant analysis, quantitative biochemistry","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of ILV budding with direct identification of Tsg101's positive regulatory role, single lab","pmids":["18768755"],"is_preprint":false},{"year":2010,"finding":"TSG101 and ubiquitin are required for cSMAC formation at the immunological synapse. siRNA knockdown of TSG101 in primary T cells impairs cSMAC formation, TCR microcluster signal termination, TCR downregulation, and segregation of TCR-MHC-peptide from PKC-theta complexes, demonstrating that ubiquitin recognition by TSG101 is a molecular checkpoint for TCR downregulation.","method":"siRNA knockdown in primary T cells, live imaging of immunological synapses, TIRF microscopy, flow cytometry","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Moderate — siRNA in primary cells with multiple defined functional readouts, single lab with orthogonal imaging and signaling assays","pmids":["20399684"],"is_preprint":false},{"year":2012,"finding":"TSG101 is recruited to the plasma membrane by binding the PSAP motif of the arrestin-domain protein ARRDC1, which drives direct plasma membrane budding to form ARRDC1-mediated microvesicles (ARMMs). This process requires VPS4 ATPase activity and is enhanced by the E3 ligase WWP2, which ubiquitinates ARRDC1.","method":"Co-immunoprecipitation, mutagenesis, live cell imaging, electron microscopy, siRNA knockdown","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with mutagenesis plus imaging and functional assays, single lab with multiple orthogonal methods","pmids":["22315426"],"is_preprint":false},{"year":2004,"finding":"TSG101 interacts with AATF and functions as a cooperative coactivator of androgen receptor-mediated transcription. TSG101 enhances monoubiquitination of the androgen receptor in a ligand-dependent manner, correlating with enhanced transactivating capacity.","method":"Co-immunoprecipitation, reporter gene assays, in vivo ubiquitination assay, dominant-negative ubiquitin overexpression","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with functional transcription assays and ubiquitination assays, single lab","pmids":["14761944"],"is_preprint":false},{"year":2002,"finding":"TSG101 binds p21(Cip1/WAF1) and increases p21 protein stability in HEK293F cells and differentiating keratinocytes. In proliferating keratinocytes, TSG101 is recruited in a p21-dependent manner to cyclin/CDK complexes and inhibits cyclin/CDK activity, causing growth suppression.","method":"Co-immunoprecipitation, pulse-chase protein stability assays, kinase activity assays, antisense RNA knockdown, cell cycle analysis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP and functional kinase assays, multiple cell types, single lab","pmids":["11943869"],"is_preprint":false},{"year":2005,"finding":"ALG-2 (a penta-EF-hand Ca2+-binding protein) directly binds the proline-rich region (PRR) of TSG101 in a Ca2+-dependent manner, as shown by GST pulldown and yeast two-hybrid. ALG-2 co-localizes with TSG101 at aberrant endosomes induced by dominant-negative SKD1/Vps4B, and this localization is Ca2+-dependent.","method":"GST pulldown, yeast two-hybrid, overlay assay with biotinylated ALG-2, immunofluorescence microscopy, deletion mapping","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — direct binding demonstrated by multiple in vitro assays with domain mapping, single lab","pmids":["16004603"],"is_preprint":false},{"year":2005,"finding":"TSG101 stabilizes the unliganded (hypophosphorylated) form of the glucocorticoid receptor (GR) by impeding its proteasomal degradation and extending its half-life. The non-phosphorylated GR (S203A/S211A mutant) shows enhanced interaction with TSG101, and siRNA depletion of TSG101 renders hypophosphorylated GR unstable.","method":"Co-immunoprecipitation, siRNA knockdown, pulse-chase analysis, proteasome inhibitor experiments, site-directed mutagenesis","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis plus functional stability assays, single lab","pmids":["15657031"],"is_preprint":false},{"year":2021,"finding":"VPS13D directly interacts with TSG101 through its adaptor-binding domain, and together they remodel lipid droplet (LD) membranes to facilitate transfer of fatty acids from LDs to mitochondria at membrane contact sites. Depletion of TSG101 or VPS13D or ESCRT-III proteins inhibits fatty acid trafficking from LDs to mitochondria.","method":"Co-immunoprecipitation, in vitro LD membrane remodeling assay, lipid transfer binding assays, siRNA knockdown, live cell imaging","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of LD membrane remodeling plus multiple siRNA knockdowns with defined functional readouts, single lab","pmids":["33623047"],"is_preprint":false},{"year":2009,"finding":"TSG101 and Hrs are required for trafficking of ubiquitylated connexin-43 (Cx43) from early endosomes to lysosomes. siRNA depletion of Hrs or TSG101 individually abrogates Cx43 trafficking to lysosomes; simultaneous depletion causes accumulation of phosphorylated and ubiquitylated Cx43 in early endosomes.","method":"siRNA knockdown, confocal microscopy, Western blotting, protein half-life measurement, microinjected Lucifer yellow transfer assays","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA with multiple orthogonal functional readouts, single lab","pmids":["19808888"],"is_preprint":false},{"year":2009,"finding":"ALG-2 bridges Alix and TSG101 as a Ca2+-dependent adaptor. ALG-2 is required for Ca2+-dependent pulldown of TSG101 by Strep-Alix; this bridge requires the ALG-2 binding site of Alix but not the PSAP motif; ALG-2 dimerization and Ca2+ binding are both required for bridging activity.","method":"Pulldown assays with purified proteins, ALG-2 knockdown, exogenous ALG-2 rescue, mutagenesis of ALG-2","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reconstitution with purified protein plus knockdown and rescue, single lab","pmids":["19520058"],"is_preprint":false},{"year":2008,"finding":"TSG101 (ESCRT-I subunit) is required for nucleocapsid release from within multivesicular endosomes to the cytoplasm during vesicular stomatitis virus (VSV) infection, specifically by controlling the back-fusion of intra-endosomal vesicles with the endosome limiting membrane, independently of its role in viral envelope fusion.","method":"siRNA knockdown, infection assays, endosomal fractionation, functional infectivity measurements","journal":"Traffic (Copenhagen, Denmark)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA with defined functional consequence and mechanistic distinction from ESCRT-0 role, single lab","pmids":["18817529"],"is_preprint":false},{"year":2020,"finding":"TSG101 interacts with SQSTM1/p62 through its PRR domain and promotes p62 aggregation, leading to recruitment of Keap1 for autophagic degradation and release of Nrf2 to the nucleus. Knockout of p62 abrogates TSG101-induced cardioprotective effects, establishing TSG101 as a regulator of the p62/Keap1/Nrf2 signaling cascade.","method":"Co-immunoprecipitation, domain deletion analysis, p62 knockout mice, transgenic and knockdown mouse models, RNA sequencing, pharmacological Nrf2 inhibition","journal":"Redox biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain mapping plus genetic epistasis in vivo, single lab","pmids":["32057709"],"is_preprint":false},{"year":2021,"finding":"TSG101 negatively regulates mitochondrial biogenesis in axons through a non-canonical, ESCRT-independent mechanism. Loss of Tsg101 activates PGC-1α/Nrf2-dependent mitochondrial biogenesis in an mTOR-independent, TFEB-dependent manner that requires the mitochondrial fission-fusion machinery. Loss of other ESCRT components does not recapitulate this phenotype.","method":"In vivo forward genetic screen (Drosophila), genetic epistasis with other ESCRT components, mitophagy and autophagy blockade, live imaging of axonal mitochondria","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo genetic screen with extensive epistasis analysis distinguishing ESCRT-independent mechanism, single lab with multiple orthogonal approaches","pmids":["33972422"],"is_preprint":false},{"year":2022,"finding":"TSG101 binds to PARP1 and is required for PARP1 activation and poly(ADP-ribose) (PAR) formation following DNA double-strand breaks. This TSG101 function is independent of its role in ESCRT-I. Absence of TSG101 impairs PAR-dependent formation of a nuclear PARP1-IKKγ signalosome, blocks IKK-NF-κB activation, causes PARP1 trapping at damage sites, and renders BRCA1/2-deficient cells synthetically lethal.","method":"Genome-wide siRNA screen, co-immunoprecipitation, PAR detection assays, NF-κB reporter assays, DNA repair assays, synthetic lethality assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — co-IP establishing direct binding plus multiple functional assays demonstrating ESCRT-independent role in DNA damage response, single lab with multiple orthogonal methods","pmids":["36124865"],"is_preprint":false},{"year":2009,"finding":"TSG101 interacts with the coiled-coil domain of GISP (G protein-coupled receptor interacting scaffold protein) and mediates lysosomal degradation of GABA(B2) receptor. GISP overexpression inhibits TSG101-dependent GABA(B2) down-regulation, and a GISP mutant lacking the TSG101-binding domain has no protective effect.","method":"Co-immunoprecipitation from rat brain, GST pulldown, deletion mapping, siRNA/overexpression in HEK293 cells, receptor degradation assays","journal":"Journal of neurochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP from native tissue plus functional receptor stability assays, single lab","pmids":["18643869"],"is_preprint":false},{"year":2009,"finding":"TSG101 interacts with multiple connexins (Cx31, Cx43, Cx45 confirmed by co-immunoprecipitation from embryonic stem cells). siRNA-mediated knockdown of TSG101 increases levels of Cx43 and Cx45, prolongs their half-life, and increases gap junction-mediated dye transfer.","method":"Yeast two-hybrid, GST pulldown, co-immunoprecipitation from HM1 ES cells, siRNA knockdown, Western blotting, Lucifer yellow transfer","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — native co-IP plus functional connexin stability assays after siRNA, single lab","pmids":["19210987"],"is_preprint":false},{"year":2012,"finding":"TSG101 interacts with the class II Rab11-FIPs (FIP3 and FIP4) through coiled-coil domains on both proteins. Point mutations in the coiled-coil regions of either TSG101 or FIP4 abrogate the interaction. Expression of TSG101 and FIP4 coiled-coil mutants causes cytokinesis defects, though the TSG101-FIP4 interaction is not required for TSG101 localization to the midbody during abscission.","method":"Proteomic screening, co-immunoprecipitation, site-directed mutagenesis, cytokinesis phenotype analysis, confocal microscopy","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP with mutagenesis plus functional cytokinesis phenotype, single lab","pmids":["22348143"],"is_preprint":false},{"year":2015,"finding":"Mahogunin (MGRN1) ubiquitinates TSG101, and this monoubiquitination is required for fusion of lysosomes with autophagosomes (via amphisomes) and late endosomes. Catalytically inactive MGRN1 or depletion of MGRN1 blocks these fusion events. Overexpression of TSG101 or its monoubiquitinated form rescues lysosomal fusion in MGRN1-deficient cells.","method":"In vivo and in vitro ubiquitination assays, siRNA knockdown, rescue by TSG101 overexpression, autophagy flux assays, lysosomal fusion assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional rescue experiments establishing the mechanistic link between TSG101 monoubiquitination and lysosomal fusion, single lab","pmids":["26539917"],"is_preprint":false},{"year":2020,"finding":"ALIX proline-rich domain (PRD) contains three tandem proline-rich motifs that compete for a single binding site on TSG101-UEV, as demonstrated by NMR relaxation dispersion and global fitting. This competitive interaction modulates ALIX function in ESCRT-mediated membrane remodeling.","method":"NMR spectroscopy (heteronuclear, relaxation dispersion), quantitative binding analysis by global fitting","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Moderate — atomic-resolution NMR with quantitative binding analysis, single lab but rigorous structural method","pmids":["32917811"],"is_preprint":false},{"year":1998,"finding":"TSG101 acts as a transcriptional suppressor through its coiled-coil domain. TSG101 represses estrogen receptor-mediated transcription and inhibits basal promoter activity; the coiled-coil domain is required for this repression and also for tumor suppressive function.","method":"Reporter gene transcription assays, deletion mutagenesis, histone acetyltransferase/deacetylase activity assays (negative result for these activities)","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional transcription assays with domain mutagenesis, single lab","pmids":["9588212"],"is_preprint":false},{"year":2023,"finding":"TSG101 interacts with HBV capsid protein HBc via the VFND motif in TSG101 and Lys-96 ubiquitination in HBc. NEDD4 (E3) and UbcH6 (E2) catalyze HBc ubiquitination required for TSG101-HBc interaction. TSG101 or NEDD4 knockdown reduces HBV particle counts in MVBs and suppresses HBV release.","method":"Co-immunoprecipitation, site-directed mutagenesis, in vitro ubiquitination assay, siRNA knockdown, transmission electron microscopy, HBV transgenic mouse model","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus site mutagenesis and functional validation in cells and in vivo, single lab with multiple orthogonal methods","pmids":["37224147"],"is_preprint":false}],"current_model":"TSG101 is a catalytically inactive ubiquitin E2 variant (UEV) that serves as the ubiquitin/PTAP-cargo-recognition subunit of the ESCRT-I complex: its UEV domain binds ubiquitylated cargo and PTAP/PSAP motifs in sorting adaptors (Hrs/HRS) and viral late domains to drive MVB/ILV biogenesis, receptor downregulation, and viral egress; it also functions independently of ESCRT-I in the nucleus (TSG101 is required for PARP1 activation and NF-κB signaling after DNA damage), in axons (suppressing PGC-1α/Nrf2-driven mitochondrial biogenesis), and at lipid droplet–mitochondria contacts (cooperating with VPS13D for fatty acid transfer), with its own steady-state levels controlled by the Tal/MGRN1-dependent monoubiquitination and proteasomal feedback loop acting through its C-terminal 'steadiness box.'"},"narrative":{"mechanistic_narrative":"TSG101 is the ubiquitin/peptide-cargo-recognition subunit of the ESCRT-I machinery that drives multivesicular body (MVB) biogenesis, receptor downregulation, and membrane scission [PMID:11208108, PMID:16707569]. Its catalytically inactive ubiquitin E2 variant (UEV) domain — structurally an E2-fold lacking the active-site cysteine and C-terminal helices — engages two spatially distinct ligands at independent sites: monoubiquitin via the Ile44 surface and PTAP/PSAP tetrapeptide motifs in a hydrophobic SH3/WW-like pocket [PMID:12006492, PMID:12379843, PMID:15053872]. Through these surfaces TSG101 recognizes ubiquitylated cargo and is recruited to endosomal membranes by the PSAP motif of the ESCRT-0 adaptor HRS, then assembles with VPS28 into the ~350 kDa ESCRT-I complex to deliver cargo such as EGFR and connexins to late endosomes/lysosomes in a VPS4-dependent manner [PMID:11134028, PMID:12802020, PMID:12900394, PMID:19808888]. This same cargo-recognition activity is usurped by enveloped viruses: the PTAP late domain of HIV-1 Gag and PTAP-like motifs of Ebola VP40 and HBV capsid directly bind the UEV domain to recruit ESCRT-I and complete budding [PMID:11595185, PMID:11726971, PMID:11427703, PMID:37224147]. TSG101 likewise mediates plasma-membrane ARMM budding via ARRDC1 and acts as a positive regulator of intraluminal vesicle formation, antagonized by ALIX, which competes for the UEV peptide-binding site and is bridged to TSG101 by Ca2+-loaded ALG-2 [PMID:18768755, PMID:22315426, PMID:32917811, PMID:19520058]. Beyond canonical sorting, TSG101 performs ESCRT-independent functions: in the nucleus it binds PARP1 and is required for PARP1 activation, NF-κB signaling after DNA damage, and synthetic lethality in BRCA1/2-deficient cells [PMID:36124865]; in axons it suppresses PGC-1α/Nrf2-driven mitochondrial biogenesis [PMID:33972422]; and at lipid droplet–mitochondria contacts it cooperates with VPS13D to remodel membranes for fatty-acid transfer [PMID:33623047]. Its own abundance is set by a C-terminal 'steadiness box' through which the E3 ligases Tal and MGRN1 monoubiquitinate TSG101, with non-ESCRT-incorporated TSG101 polyubiquitinated for proteasomal degradation unless protected by VPS28/ESCRT-I assembly [PMID:15256501, PMID:17229889, PMID:18077552]. TSG101 loss in vivo stabilizes p53, and a p53-null background rescues the gastrulation defect of TSG101-null embryos, linking it genetically to the p53 pathway [PMID:11172041].","teleology":[{"year":1998,"claim":"Established TSG101's cell-cycle-dependent subcellular distribution and a role in genome stability, raising the question of its biochemical activity.","evidence":"Immunofluorescence across the cell cycle and phenotyping of deficient cells","pmids":["9465061"],"confidence":"Medium","gaps":["No molecular mechanism for spindle/centrosome function","Relationship to later ESCRT role unresolved"]},{"year":2000,"claim":"Identified TSG101 as a VPS23 ortholog acting in late-endosomal cargo delivery and showed its abundance is set posttranslationally by a C-terminal steadiness box, defining its trafficking role and an autoregulatory layer.","evidence":"Trafficking assays in mutant cells; deletion mapping and pulse-chase; co-IP/cross-linking with VPS28","pmids":["11208108","10749147","11134028"],"confidence":"High","gaps":["Steadiness-box ligase not yet identified","Mechanism of cargo selection not yet structural"]},{"year":2001,"claim":"Defined the UEV domain as a catalytically inactive E2 variant that binds ubiquitin and PTAP late domains, establishing TSG101/ESCRT-I as the cellular machinery hijacked for HIV-1 and Ebola budding.","evidence":"siRNA depletion with rescue, binding assays, dominant-negative VPS4, EM; site-directed mutagenesis of Tyr-110","pmids":["11595185","11726971","11427703"],"confidence":"High","gaps":["Atomic basis of dual-ligand recognition not yet solved","Composition of full ESCRT-I in budding incomplete"]},{"year":2001,"claim":"Linked TSG101 to the p53/MDM2 axis in vivo, showing its loss stabilizes p53 and that p53 ablation rescues the null embryonic phenotype.","evidence":"Knockout and double-knockout mice with genetic epistasis; MDM2/p53 biochemistry","pmids":["11172041","11172000"],"confidence":"High","gaps":["Direct mechanism of MDM2 ubiquitination interference not reconstituted","Whether this is ESCRT-dependent unclear"]},{"year":2002,"claim":"Solved UEV-domain structures alone, with PTAP peptide, establishing distinct, independent binding pockets for peptide ligands and the vestigial active site.","evidence":"NMR structures with SPR/chemical-shift mapping","pmids":["12006492","12379843"],"confidence":"High","gaps":["Ubiquitin-binding geometry not yet co-crystallized (resolved later)"]},{"year":2003,"claim":"Identified HRS as the physiological PSAP adaptor recruiting TSG101 to endosomes, explaining how cellular cargo sorting and viral budding share the same recruitment logic.","evidence":"Reciprocal co-IP, yeast two-hybrid, functional rescue and EGFR trafficking assays","pmids":["12802020","12900394"],"confidence":"High","gaps":["Stoichiometry of HRS-TSG101 handoff unclear","How ubiquitin and PSAP binding are coordinated unresolved"]},{"year":2004,"claim":"Determined the UEV-ubiquitin co-crystal structure and identified Tal as an E3 ligase that monoubiquitinates TSG101, defining both the ubiquitin-recognition interface and a regulatory ubiquitination input.","evidence":"X-ray crystallography with mutagenesis/MVB assays; in vitro ubiquitination and domain mapping","pmids":["15053872","15256501"],"confidence":"High","gaps":["Functional consequence of monoubiquitination not fully defined here","Whether Tal acts on assembled vs free TSG101 unclear"]},{"year":2006,"claim":"Distinguished TSG101's role in MVB formation itself from HRS's role in ILV accumulation, separating ESCRT-I and ESCRT-0 functions.","evidence":"siRNA with EM and EGF-degradation assays","pmids":["16707569"],"confidence":"High","gaps":["Biophysical mechanism of membrane deformation not addressed"]},{"year":2007,"claim":"Showed MGRN1 (Mahogunin) monoubiquitinates TSG101 and that the steadiness box governs Tal-mediated polyubiquitination/degradation gated by VPS28 availability, defining how ESCRT-I assembly protects TSG101.","evidence":"In vitro ubiquitination, proteasome inhibition, siRNA, co-IP","pmids":["17229889","18077552"],"confidence":"High","gaps":["Switch between mono- and poly-ubiquitination not fully resolved","In vivo relevance of feedback loop untested"]},{"year":2008,"claim":"Reconstituted ILV budding in vitro and assigned TSG101 a positive regulatory role antagonized by ALIX; extended ESCRT-I function to viral nucleocapsid back-fusion.","evidence":"Cell-free budding reconstitution; VSV infection and endosomal fractionation","pmids":["18768755","18817529"],"confidence":"Medium","gaps":["Molecular basis of ALIX antagonism not yet structural (resolved 2020)","Back-fusion mechanism single-lab"]},{"year":2012,"claim":"Generalized TSG101 recruitment beyond endosomes to direct plasma-membrane ARMM budding via ARRDC1 and to midbody/cytokinesis via Rab11-FIP coiled-coil interactions.","evidence":"Co-IP, mutagenesis, live imaging, EM; cytokinesis phenotyping","pmids":["22315426","22348143"],"confidence":"Medium","gaps":["FIP4 interaction dispensable for midbody localization, leaving recruitment mechanism open","ARMM cargo selectivity unclear"]},{"year":2020,"claim":"Resolved how ALIX competes with itself and TSG101 at the UEV peptide site and uncovered a p62/Keap1/Nrf2 cardioprotective signaling role, broadening TSG101 function.","evidence":"NMR relaxation dispersion; co-IP, domain mapping, p62-knockout mice","pmids":["32917811","32057709"],"confidence":"Medium","gaps":["Physiological balance of ALIX vs cargo competition in cells unmeasured","Direct vs indirect p62 effects not fully separated"]},{"year":2021,"claim":"Defined two ESCRT-independent functions: lipid-droplet-to-mitochondria fatty-acid transfer with VPS13D, and suppression of axonal mitochondrial biogenesis via PGC-1α/Nrf2.","evidence":"In vitro LD remodeling and lipid transfer; Drosophila forward genetic screen with ESCRT epistasis","pmids":["33623047","33972422"],"confidence":"High","gaps":["How a single protein partitions between ESCRT and non-ESCRT pools unknown","Direct biochemical activity in mitochondrial biogenesis suppression undefined"]},{"year":2022,"claim":"Established a nuclear, ESCRT-independent role in which TSG101 binds and activates PARP1 to drive NF-κB signaling and confers synthetic lethality in BRCA1/2-deficient cells.","evidence":"Genome-wide siRNA screen, co-IP, PAR and NF-κB assays, synthetic-lethality assays","pmids":["36124865"],"confidence":"High","gaps":["How TSG101 mechanistically promotes PARP1 catalysis unresolved","Nuclear targeting signal/regulation not defined"]},{"year":2023,"claim":"Extended viral-egress recruitment to HBV, requiring NEDD4/UbcH6-catalyzed ubiquitination of the HBc capsid and a VFND motif in TSG101.","evidence":"In vitro ubiquitination, mutagenesis, siRNA, EM, transgenic mouse","pmids":["37224147"],"confidence":"High","gaps":["Structural basis of VFND-mediated contact undefined","Whether full ESCRT-I is required for HBV release untested"]},{"year":null,"claim":"How TSG101 is partitioned and regulated between ESCRT-I-bound cytoplasmic pools and its ESCRT-independent nuclear, axonal, and membrane-contact-site roles remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No mechanism for nuclear import/retention","No structure of full ESCRT-I human complex in the timeline","Determinants directing TSG101 to non-ESCRT functions unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0031386","term_label":"protein tag activity","supporting_discovery_ids":[0,4,6]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[8,9,21]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[10,25,32]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[26]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[3,7,18]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[7]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[21]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[17,32]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[17]},{"term_id":"GO:0005811","term_label":"lipid droplet","supporting_discovery_ids":[26]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[3,8,18,19]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[0,1,39]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[32]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[30,36]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[13,14,15]}],"complexes":["ESCRT-I"],"partners":["VPS28","HRS","ALIX","ALG-2","ARRDC1","VPS13D","PARP1","MGRN1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q99816","full_name":"Tumor susceptibility gene 101 protein","aliases":["ESCRT-I complex subunit TSG101"],"length_aa":390,"mass_kda":43.9,"function":"Component of the ESCRT-I complex, a regulator of vesicular trafficking process. Binds to ubiquitinated cargo proteins and is required for the sorting of endocytic ubiquitinated cargos into multivesicular bodies (MVBs). Mediates the association between the ESCRT-0 and ESCRT-I complex. Required for completion of cytokinesis; the function requires CEP55. May be involved in cell growth and differentiation. Acts as a negative growth regulator. Involved in the budding of many viruses through an interaction with viral proteins that contain a late-budding motif P-[ST]-A-P. This interaction is essential for viral particle budding of numerous retroviruses. Required for the exosomal release of SDCBP, CD63 and syndecan (PubMed:22660413). It may also play a role in the extracellular release of microvesicles that differ from the exosomes (PubMed:22315426)","subcellular_location":"Cytoplasm; Early endosome membrane; Late endosome membrane; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Midbody, Midbody ring; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q99816/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/TSG101","classification":"Common Essential","n_dependent_lines":1176,"n_total_lines":1208,"dependency_fraction":0.9735099337748344},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000074319","cell_line_id":"CID000787","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"nucleoplasm","grade":1}],"interactors":[{"gene":"MVB12A","stoichiometry":10.0},{"gene":"UMAD1","stoichiometry":10.0},{"gene":"VPS28","stoichiometry":10.0},{"gene":"VPS37A","stoichiometry":10.0},{"gene":"VPS37B","stoichiometry":10.0},{"gene":"UBAP1","stoichiometry":10.0},{"gene":"STAM2","stoichiometry":0.2},{"gene":"HGS","stoichiometry":0.2},{"gene":"LAMC1","stoichiometry":0.2},{"gene":"VPS37C","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000787","total_profiled":1310},"omim":[{"mim_id":"621454","title":"MULTIVESICULAR BODY SUBUNIT 12B; MVB12B","url":"https://www.omim.org/entry/621454"},{"mim_id":"621453","title":"MULTIVESICULAR BODY SUBUNIT 12A; MVB12A","url":"https://www.omim.org/entry/621453"},{"mim_id":"619788","title":"ARRESTIN DOMAIN-CONTAINING PROTEIN 4; ARRDC4","url":"https://www.omim.org/entry/619788"},{"mim_id":"619768","title":"ARRESTIN DOMAIN-CONTAINING PROTEIN 1; ARRDC1","url":"https://www.omim.org/entry/619768"},{"mim_id":"618627","title":"GERM CELL-LESS 1, SPERMATOGENESIS-ASSOCIATED; GMCL1","url":"https://www.omim.org/entry/618627"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Nucleoli","reliability":"Additional"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/TSG101"},"hgnc":{"alias_symbol":["VPS23"],"prev_symbol":["TSG10"]},"alphafold":{"accession":"Q99816","domains":[{"cath_id":"3.10.110.10","chopping":"4-142","consensus_level":"high","plddt":96.1463,"start":4,"end":142},{"cath_id":"1.10.287","chopping":"319-383","consensus_level":"high","plddt":92.8688,"start":319,"end":383}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99816","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q99816-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q99816-F1-predicted_aligned_error_v6.png","plddt_mean":82.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TSG101","jax_strain_url":"https://www.jax.org/strain/search?query=TSG101"},"sequence":{"accession":"Q99816","fasta_url":"https://rest.uniprot.org/uniprotkb/Q99816.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q99816/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99816"}},"corpus_meta":[{"pmid":"11595185","id":"PMC_11595185","title":"Tsg101 and the vacuolar protein sorting pathway are essential for HIV-1 budding.","date":"2001","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/11595185","citation_count":1169,"is_preprint":false},{"pmid":"11726971","id":"PMC_11726971","title":"HIV-1 and Ebola virus encode small peptide motifs that recruit Tsg101 to sites of particle assembly to facilitate egress.","date":"2001","source":"Nature medicine","url":"https://pubmed.ncbi.nlm.nih.gov/11726971","citation_count":615,"is_preprint":false},{"pmid":"22315426","id":"PMC_22315426","title":"Formation and release of arrestin domain-containing protein 1-mediated microvesicles (ARMMs) at plasma membrane by recruitment of TSG101 protein.","date":"2012","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/22315426","citation_count":580,"is_preprint":false},{"pmid":"11427703","id":"PMC_11427703","title":"Tsg101, a homologue of ubiquitin-conjugating (E2) enzymes, binds the L domain in HIV type 1 Pr55(Gag).","date":"2001","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/11427703","citation_count":508,"is_preprint":false},{"pmid":"11208108","id":"PMC_11208108","title":"Mammalian tumor susceptibility gene 101 (TSG101) and the yeast homologue, Vps23p, both function in late endosomal trafficking.","date":"2000","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/11208108","citation_count":360,"is_preprint":false},{"pmid":"8616888","id":"PMC_8616888","title":"Tsg101: a novel tumor susceptibility gene isolated by controlled homozygous functional knockout of allelic loci in mammalian cells.","date":"1996","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/8616888","citation_count":310,"is_preprint":false},{"pmid":"11805336","id":"PMC_11805336","title":"Overexpression of the N-terminal domain of TSG101 inhibits HIV-1 budding by blocking late domain function.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/11805336","citation_count":307,"is_preprint":false},{"pmid":"12802020","id":"PMC_12802020","title":"TSG101 interaction with HRS mediates endosomal trafficking and receptor down-regulation.","date":"2003","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12802020","citation_count":274,"is_preprint":false},{"pmid":"12006492","id":"PMC_12006492","title":"Structure and functional interactions of the Tsg101 UEV domain.","date":"2002","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/12006492","citation_count":258,"is_preprint":false},{"pmid":"12525615","id":"PMC_12525615","title":"Overlapping motifs (PTAP and PPEY) within the Ebola virus VP40 protein function independently as late budding domains: involvement of host proteins TSG101 and VPS-4.","date":"2003","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/12525615","citation_count":234,"is_preprint":false},{"pmid":"12900394","id":"PMC_12900394","title":"HIV Gag mimics the Tsg101-recruiting activity of the human Hrs protein.","date":"2003","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/12900394","citation_count":215,"is_preprint":false},{"pmid":"12379843","id":"PMC_12379843","title":"Structure of the Tsg101 UEV domain in complex with the PTAP motif of the HIV-1 p6 protein.","date":"2002","source":"Nature structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/12379843","citation_count":209,"is_preprint":false},{"pmid":"16707569","id":"PMC_16707569","title":"Distinct roles for Tsg101 and Hrs in multivesicular body formation and inward vesiculation.","date":"2006","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/16707569","citation_count":207,"is_preprint":false},{"pmid":"15053872","id":"PMC_15053872","title":"Ubiquitin recognition by the human TSG101 protein.","date":"2004","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/15053872","citation_count":182,"is_preprint":false},{"pmid":"11134028","id":"PMC_11134028","title":"TSG101/mammalian VPS23 and mammalian VPS28 interact directly and are recruited to VPS4-induced endosomes.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11134028","citation_count":170,"is_preprint":false},{"pmid":"12006494","id":"PMC_12006494","title":"Ubiquitylation of MHC class I by the K3 viral protein signals internalization and TSG101-dependent degradation.","date":"2002","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/12006494","citation_count":170,"is_preprint":false},{"pmid":"12559917","id":"PMC_12559917","title":"Ebola virus matrix protein VP40 interaction with human cellular factors Tsg101 and Nedd4.","date":"2003","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12559917","citation_count":161,"is_preprint":false},{"pmid":"16014378","id":"PMC_16014378","title":"Depletion of TSG101 forms a mammalian \"Class E\" compartment: a multicisternal early endosome with multiple sorting defects.","date":"2005","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/16014378","citation_count":156,"is_preprint":false},{"pmid":"11172000","id":"PMC_11172000","title":"A TSG101/MDM2 regulatory loop modulates MDM2 degradation and MDM2/p53 feedback control.","date":"2001","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/11172000","citation_count":147,"is_preprint":false},{"pmid":"17982468","id":"PMC_17982468","title":"Beyond Tsg101: the role of Alix in 'ESCRTing' HIV-1.","date":"2007","source":"Nature reviews. Microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/17982468","citation_count":142,"is_preprint":false},{"pmid":"11172041","id":"PMC_11172041","title":"p53 accumulation, defective cell proliferation, and early embryonic lethality in mice lacking tsg101.","date":"2001","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/11172041","citation_count":137,"is_preprint":false},{"pmid":"20399684","id":"PMC_20399684","title":"Essential role of ubiquitin and TSG101 protein in formation and function of the central supramolecular activation cluster.","date":"2010","source":"Immunity","url":"https://pubmed.ncbi.nlm.nih.gov/20399684","citation_count":133,"is_preprint":false},{"pmid":"15256501","id":"PMC_15256501","title":"Tal, a Tsg101-specific E3 ubiquitin ligase, regulates receptor endocytosis and retrovirus budding.","date":"2004","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/15256501","citation_count":130,"is_preprint":false},{"pmid":"9019400","id":"PMC_9019400","title":"The TSG101 tumor susceptibility gene is located in chromosome 11 band p15 and is mutated in human breast cancer.","date":"1997","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/9019400","citation_count":130,"is_preprint":false},{"pmid":"15126635","id":"PMC_15126635","title":"Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding.","date":"2004","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/15126635","citation_count":128,"is_preprint":false},{"pmid":"19692479","id":"PMC_19692479","title":"Herpes simplex virus type 1 production requires a functional ESCRT-III complex but is independent of TSG101 and ALIX expression.","date":"2009","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/19692479","citation_count":127,"is_preprint":false},{"pmid":"17229889","id":"PMC_17229889","title":"Spongiform neurodegeneration-associated E3 ligase Mahogunin ubiquitylates TSG101 and regulates endosomal trafficking.","date":"2007","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/17229889","citation_count":121,"is_preprint":false},{"pmid":"19053244","id":"PMC_19053244","title":"Inhibition of HIV budding by a genetically selected cyclic peptide targeting the Gag-TSG101 interaction.","date":"2008","source":"ACS chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/19053244","citation_count":121,"is_preprint":false},{"pmid":"12482969","id":"PMC_12482969","title":"Tsg101 is essential for cell growth, proliferation, and cell survival of embryonic and adult tissues.","date":"2003","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12482969","citation_count":119,"is_preprint":false},{"pmid":"16407403","id":"PMC_16407403","title":"An essential role for SNX1 in lysosomal sorting of protease-activated receptor-1: evidence for retromer-, Hrs-, and Tsg101-independent functions of sorting nexins.","date":"2006","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/16407403","citation_count":113,"is_preprint":false},{"pmid":"33623047","id":"PMC_33623047","title":"An ESCRT-dependent step in fatty acid transfer from lipid droplets to mitochondria through VPS13D-TSG101 interactions.","date":"2021","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33623047","citation_count":113,"is_preprint":false},{"pmid":"14581525","id":"PMC_14581525","title":"PPPYVEPTAP motif is the late domain of human T-cell leukemia virus type 1 Gag and mediates its functional interaction with cellular proteins Nedd4 and Tsg101 [corrected].","date":"2003","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/14581525","citation_count":113,"is_preprint":false},{"pmid":"16352570","id":"PMC_16352570","title":"Nonstructural protein 3 of bluetongue virus assists virus release by recruiting ESCRT-I protein Tsg101.","date":"2006","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/16352570","citation_count":112,"is_preprint":false},{"pmid":"9465061","id":"PMC_9465061","title":"Cell cycle-dependent subcellular localization of the TSG101 protein and mitotic and nuclear abnormalities associated with TSG101 deficiency.","date":"1998","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/9465061","citation_count":101,"is_preprint":false},{"pmid":"17301151","id":"PMC_17301151","title":"Interaction of Tsg101 with Marburg virus VP40 depends on the PPPY motif, but not the PT/SAP motif as in the case of Ebola virus, and Tsg101 plays a critical role in the budding of Marburg virus-like particles induced by VP40, NP, and GP.","date":"2007","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/17301151","citation_count":91,"is_preprint":false},{"pmid":"18768755","id":"PMC_18768755","title":"In vitro budding of intralumenal vesicles into late endosomes is regulated by Alix and Tsg101.","date":"2008","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18768755","citation_count":89,"is_preprint":false},{"pmid":"9366528","id":"PMC_9366528","title":"Aberrant splicing of the TSG101 and FHIT genes occurs frequently in multiple malignancies and in normal tissues and mimics alterations previously described in tumours.","date":"1997","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9366528","citation_count":86,"is_preprint":false},{"pmid":"19808888","id":"PMC_19808888","title":"Ubiquitylation of the gap junction protein connexin-43 signals its trafficking from early endosomes to lysosomes in a process mediated by Hrs and Tsg101.","date":"2009","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/19808888","citation_count":86,"is_preprint":false},{"pmid":"12743307","id":"PMC_12743307","title":"Defects in human immunodeficiency virus budding and endosomal sorting induced by TSG101 overexpression.","date":"2003","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/12743307","citation_count":86,"is_preprint":false},{"pmid":"9253709","id":"PMC_9253709","title":"The breast cancer gene product TSG101: a regulator of ubiquitination?","date":"1997","source":"Journal of molecular medicine (Berlin, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/9253709","citation_count":85,"is_preprint":false},{"pmid":"14761944","id":"PMC_14761944","title":"TSG101 interacts with apoptosis-antagonizing transcription factor and enhances androgen receptor-mediated transcription by promoting its monoubiquitination.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14761944","citation_count":84,"is_preprint":false},{"pmid":"12915533","id":"PMC_12915533","title":"Tsg101 control of human immunodeficiency virus type 1 Gag trafficking and release.","date":"2003","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/12915533","citation_count":81,"is_preprint":false},{"pmid":"12205095","id":"PMC_12205095","title":"Targeted deletion of the Tsg101 gene results in cell cycle arrest at G1/S and p53-independent cell death.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12205095","citation_count":77,"is_preprint":false},{"pmid":"12101421","id":"PMC_12101421","title":"Overexpression of tumor susceptibility gene TSG101 in human papillary thyroid carcinomas.","date":"2002","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12101421","citation_count":70,"is_preprint":false},{"pmid":"16004603","id":"PMC_16004603","title":"The penta-EF-hand protein ALG-2 interacts directly with the ESCRT-I component TSG101, and Ca2+-dependently co-localizes to aberrant endosomes with dominant-negative AAA ATPase SKD1/Vps4B.","date":"2005","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/16004603","citation_count":69,"is_preprint":false},{"pmid":"23132495","id":"PMC_23132495","title":"Cryptococcus neoformans requires the ESCRT protein Vps23 for iron acquisition from heme, for capsule formation, and for virulence.","date":"2012","source":"Infection and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/23132495","citation_count":66,"is_preprint":false},{"pmid":"26066081","id":"PMC_26066081","title":"WASH and Tsg101/ALIX-dependent diversion of stress-internalized EGFR from the canonical endocytic pathway.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26066081","citation_count":66,"is_preprint":false},{"pmid":"18077552","id":"PMC_18077552","title":"Regulation of Tsg101 expression by the steadiness box: a role of Tsg101-associated ligase.","date":"2007","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18077552","citation_count":64,"is_preprint":false},{"pmid":"16118794","id":"PMC_16118794","title":"SIMPLE interacts with NEDD4 and TSG101: evidence for a role in lysosomal sorting and implications for Charcot-Marie-Tooth disease.","date":"2005","source":"Journal of neuroscience research","url":"https://pubmed.ncbi.nlm.nih.gov/16118794","citation_count":64,"is_preprint":false},{"pmid":"15210712","id":"PMC_15210712","title":"Cell cycle arrest and cell death are controlled by p53-dependent and p53-independent mechanisms in Tsg101-deficient cells.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15210712","citation_count":58,"is_preprint":false},{"pmid":"10749147","id":"PMC_10749147","title":"TSG101 protein steady-state level is regulated posttranslationally by an evolutionarily conserved COOH-terminal sequence.","date":"2000","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/10749147","citation_count":57,"is_preprint":false},{"pmid":"18817529","id":"PMC_18817529","title":"The ESCRT-I subunit TSG101 controls endosome-to-cytosol release of viral RNA.","date":"2008","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/18817529","citation_count":54,"is_preprint":false},{"pmid":"11943869","id":"PMC_11943869","title":"Negative regulation of cell growth and differentiation by TSG101 through association with p21(Cip1/WAF1).","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/11943869","citation_count":52,"is_preprint":false},{"pmid":"17369844","id":"PMC_17369844","title":"Tsg101 is upregulated in a subset of invasive human breast cancers and its targeted overexpression in transgenic mice reveals weak oncogenic properties for mammary cancer initiation.","date":"2007","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/17369844","citation_count":52,"is_preprint":false},{"pmid":"9588212","id":"PMC_9588212","title":"A putative tumor suppressor, TSG101, acts as a transcriptional suppressor through its coiled-coil domain.","date":"1998","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/9588212","citation_count":51,"is_preprint":false},{"pmid":"13679614","id":"PMC_13679614","title":"Association of Japanese encephalitis virus NS3 protein with microtubules and tumour susceptibility gene 101 (TSG101) protein.","date":"2003","source":"The Journal of general virology","url":"https://pubmed.ncbi.nlm.nih.gov/13679614","citation_count":50,"is_preprint":false},{"pmid":"25330247","id":"PMC_25330247","title":"Interaction with Tsg101 is necessary for the efficient transport and release of nucleocapsids in marburg virus-infected cells.","date":"2014","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/25330247","citation_count":50,"is_preprint":false},{"pmid":"9661875","id":"PMC_9661875","title":"Perturbation of TSG101 protein affects cell cycle progression.","date":"1998","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/9661875","citation_count":49,"is_preprint":false},{"pmid":"14991575","id":"PMC_14991575","title":"Reduction of TSG101 protein has a negative impact on tumor cell growth.","date":"2004","source":"International journal of cancer","url":"https://pubmed.ncbi.nlm.nih.gov/14991575","citation_count":49,"is_preprint":false},{"pmid":"19520058","id":"PMC_19520058","title":"Penta-EF-hand protein ALG-2 functions as a Ca2+-dependent adaptor that bridges Alix and TSG101.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19520058","citation_count":47,"is_preprint":false},{"pmid":"32057709","id":"PMC_32057709","title":"Tsg101 positively regulates P62-Keap1-Nrf2 pathway to protect hearts against oxidative damage.","date":"2020","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/32057709","citation_count":46,"is_preprint":false},{"pmid":"11973141","id":"PMC_11973141","title":"Tsg101: HIV-1's ticket to ride.","date":"2002","source":"Trends in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/11973141","citation_count":46,"is_preprint":false},{"pmid":"9444960","id":"PMC_9444960","title":"Frequent abnormalities of TSG101 transcripts in human prostate cancer.","date":"1997","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9444960","citation_count":46,"is_preprint":false},{"pmid":"19703557","id":"PMC_19703557","title":"Abnormal regulation of TSG101 in mice with spongiform neurodegeneration.","date":"2009","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/19703557","citation_count":45,"is_preprint":false},{"pmid":"16138902","id":"PMC_16138902","title":"The functionally exchangeable L domains in RSV and HIV-1 Gag direct particle release through pathways linked by Tsg101.","date":"2005","source":"Traffic (Copenhagen, Denmark)","url":"https://pubmed.ncbi.nlm.nih.gov/16138902","citation_count":44,"is_preprint":false},{"pmid":"16571793","id":"PMC_16571793","title":"Vpu and Tsg101 regulate intracellular targeting of the human immunodeficiency virus type 1 core protein precursor Pr55gag.","date":"2006","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/16571793","citation_count":43,"is_preprint":false},{"pmid":"15657031","id":"PMC_15657031","title":"Stabilization of the unliganded glucocorticoid receptor by TSG101.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15657031","citation_count":41,"is_preprint":false},{"pmid":"26539917","id":"PMC_26539917","title":"Mahogunin regulates fusion between amphisomes/MVBs and lysosomes via ubiquitination of TSG101.","date":"2015","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/26539917","citation_count":39,"is_preprint":false},{"pmid":"17940959","id":"PMC_17940959","title":"Involvement of vacuolar protein sorting pathway in Ebola virus release independent of TSG101 interaction.","date":"2007","source":"The Journal of infectious diseases","url":"https://pubmed.ncbi.nlm.nih.gov/17940959","citation_count":38,"is_preprint":false},{"pmid":"9840940","id":"PMC_9840940","title":"Genomic architecture and transcriptional activation of the mouse and human tumor susceptibility gene TSG101: common types of shorter transcripts are true alternative splice variants.","date":"1998","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9840940","citation_count":38,"is_preprint":false},{"pmid":"17182691","id":"PMC_17182691","title":"Role of the TSG101 gene in Epstein-Barr virus late gene transcription.","date":"2006","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/17182691","citation_count":37,"is_preprint":false},{"pmid":"19210987","id":"PMC_19210987","title":"The TSG101 protein binds to connexins and is involved in connexin degradation.","date":"2009","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/19210987","citation_count":36,"is_preprint":false},{"pmid":"27203423","id":"PMC_27203423","title":"Nipah Virus C Protein Recruits Tsg101 to Promote the Efficient Release of Virus in an ESCRT-Dependent Pathway.","date":"2016","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/27203423","citation_count":36,"is_preprint":false},{"pmid":"32075127","id":"PMC_32075127","title":"The Multifaceted Roles of the Tumor Susceptibility Gene 101 (TSG101) in Normal Development and Disease.","date":"2020","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/32075127","citation_count":34,"is_preprint":false},{"pmid":"36076942","id":"PMC_36076942","title":"The Neuroprotective Effects of Exosomes Derived from TSG101-Overexpressing Human Neural Stem Cells in a Stroke Model.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/36076942","citation_count":34,"is_preprint":false},{"pmid":"27764233","id":"PMC_27764233","title":"ESCRT-I Protein Tsg101 Plays a Role in the Post-macropinocytic Trafficking and Infection of Endothelial Cells by Kaposi's Sarcoma-Associated Herpesvirus.","date":"2016","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/27764233","citation_count":33,"is_preprint":false},{"pmid":"31390267","id":"PMC_31390267","title":"Circadian variation in the release of small extracellular vesicles can be normalized by vesicle number or TSG101.","date":"2019","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/31390267","citation_count":33,"is_preprint":false},{"pmid":"37224147","id":"PMC_37224147","title":"Hepatitis B virus hijacks TSG101 to facilitate egress via multiple vesicle bodies.","date":"2023","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/37224147","citation_count":31,"is_preprint":false},{"pmid":"20088757","id":"PMC_20088757","title":"TSG101: a novel anti-HIV-1 drug target.","date":"2010","source":"Current medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20088757","citation_count":30,"is_preprint":false},{"pmid":"18555885","id":"PMC_18555885","title":"Tsg101 can replace Nedd4 function in ASV Gag release but not membrane targeting.","date":"2008","source":"Virology","url":"https://pubmed.ncbi.nlm.nih.gov/18555885","citation_count":30,"is_preprint":false},{"pmid":"24672030","id":"PMC_24672030","title":"A unique N-terminal sequence in the Carnation Italian ringspot virus p36 replicase-associated protein interacts with the host cell ESCRT-I component Vps23.","date":"2014","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/24672030","citation_count":29,"is_preprint":false},{"pmid":"21117030","id":"PMC_21117030","title":"Down-regulation of TSG101 by small interfering RNA inhibits the proliferation of breast cancer cells through the MAPK/ERK signal pathway.","date":"2011","source":"Histology and histopathology","url":"https://pubmed.ncbi.nlm.nih.gov/21117030","citation_count":29,"is_preprint":false},{"pmid":"32917811","id":"PMC_32917811","title":"Proline-rich domain of human ALIX contains multiple TSG101-UEV interaction sites and forms phosphorylation-mediated reversible amyloids.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32917811","citation_count":28,"is_preprint":false},{"pmid":"23933150","id":"PMC_23933150","title":"RAB7 and TSG101 are required for the constitutive recycling of unliganded EGFRs via distinct mechanisms.","date":"2013","source":"Molecular and cellular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/23933150","citation_count":27,"is_preprint":false},{"pmid":"33328308","id":"PMC_33328308","title":"The ESCRT-I Subunit Tsg101 Plays Novel Dual Roles in Entry and Replication of Classical Swine Fever Virus.","date":"2021","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/33328308","citation_count":27,"is_preprint":false},{"pmid":"17182674","id":"PMC_17182674","title":"The C-terminal portion of the Hrs protein interacts with Tsg101 and interferes with human immunodeficiency virus type 1 Gag particle production.","date":"2006","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/17182674","citation_count":27,"is_preprint":false},{"pmid":"18600204","id":"PMC_18600204","title":"Overexpression of WNT2 and TSG101 genes in colorectal carcinoma.","date":"2008","source":"Tropical biomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/18600204","citation_count":26,"is_preprint":false},{"pmid":"25488808","id":"PMC_25488808","title":"HIV-1 nucleocapsid and ESCRT-component Tsg101 interplay prevents HIV from turning into a DNA-containing virus.","date":"2014","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/25488808","citation_count":26,"is_preprint":false},{"pmid":"22348143","id":"PMC_22348143","title":"Tumor susceptibility gene 101 (TSG101) is a novel binding-partner for the class II Rab11-FIPs.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22348143","citation_count":26,"is_preprint":false},{"pmid":"36124865","id":"PMC_36124865","title":"TSG101 associates with PARP1 and is essential for PARylation and DNA damage-induced NF-κB activation.","date":"2022","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/36124865","citation_count":25,"is_preprint":false},{"pmid":"18643869","id":"PMC_18643869","title":"GISP binding to TSG101 increases GABA receptor stability by down-regulating ESCRT-mediated lysosomal degradation.","date":"2008","source":"Journal of neurochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18643869","citation_count":25,"is_preprint":false},{"pmid":"32895884","id":"PMC_32895884","title":"MiR-17-5p-mediated endoplasmic reticulum stress promotes acute myocardial ischemia injury through targeting Tsg101.","date":"2020","source":"Cell stress & chaperones","url":"https://pubmed.ncbi.nlm.nih.gov/32895884","citation_count":25,"is_preprint":false},{"pmid":"30450735","id":"PMC_30450735","title":"TSG101 promotes the proliferation, migration and invasion of hepatocellular carcinoma cells by regulating the PEG10.","date":"2018","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/30450735","citation_count":24,"is_preprint":false},{"pmid":"33972422","id":"PMC_33972422","title":"TSG101 negatively regulates mitochondrial biogenesis in axons.","date":"2021","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/33972422","citation_count":23,"is_preprint":false},{"pmid":"9331081","id":"PMC_9331081","title":"Identification of cellular TSG101 protein in multiple human breast cancer cell lines.","date":"1997","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/9331081","citation_count":23,"is_preprint":false},{"pmid":"23276921","id":"PMC_23276921","title":"Role of TSG101 in cancer.","date":"2013","source":"Frontiers in bioscience (Landmark edition)","url":"https://pubmed.ncbi.nlm.nih.gov/23276921","citation_count":22,"is_preprint":false},{"pmid":"22479596","id":"PMC_22479596","title":"A knockout of the Tsg101 gene leads to decreased expression of ErbB receptor tyrosine kinases and induction of autophagy prior to cell death.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22479596","citation_count":22,"is_preprint":false},{"pmid":"19542561","id":"PMC_19542561","title":"Proline-rich sequence recognition: II. Proteomics analysis of Tsg101 ubiquitin-E2-like variant (UEV) interactions.","date":"2009","source":"Molecular & cellular proteomics : MCP","url":"https://pubmed.ncbi.nlm.nih.gov/19542561","citation_count":22,"is_preprint":false},{"pmid":"15033475","id":"PMC_15033475","title":"Physical and functional interactions between Daxx and TSG101.","date":"2004","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/15033475","citation_count":21,"is_preprint":false},{"pmid":"9773405","id":"PMC_9773405","title":"Aberrant splicing of the TSG101 tumor suppressor gene in human breast and ovarian cancers.","date":"1998","source":"Journal of the Society for Gynecologic Investigation","url":"https://pubmed.ncbi.nlm.nih.gov/9773405","citation_count":21,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":47257,"output_tokens":9709,"usd":0.143703,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":20057,"output_tokens":5132,"usd":0.114293,"stage2_stop_reason":"end_turn"},"total_usd":0.257996,"stage1_batch_id":"msgbatch_017qqVz33Ga63SCDwzb35XPb","stage2_batch_id":"msgbatch_01Scr2Ei9x4HJMiRGxwrmvLK","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2001,\n      \"finding\": \"The UEV (ubiquitin E2 variant) domain of TSG101 directly binds the PTAP tetrapeptide 'late domain' motif of HIV-1 Gag p6 protein, and also binds ubiquitin. Depletion of TSG101 by siRNA arrests HIV-1 budding at a late stage; reintroduction rescues budding. Dominant-negative VPS4 also arrests HIV-1 budding, placing TSG101 in the vacuolar protein sorting (VPS)/ESCRT pathway required for HIV-1 egress.\",\n      \"method\": \"siRNA depletion, co-immunoprecipitation, surface plasmon resonance binding assay, dominant-negative VPS4 overexpression, electron microscopy\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal methods (siRNA, binding assays, dominant-negative, EM), replicated across multiple subsequent studies\",\n      \"pmids\": [\"11595185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"TSG101 interacts with the PTAP-containing late domain of HIV-1 Gag and the PTAP-like motif of Ebola VP40 (EbVp40). Recruitment of TSG101 to assembling virions restores budding competence to late-domain-defective HIV-1, establishing that TSG101 recruitment is the essential function of the viral late domain.\",\n      \"method\": \"Co-immunoprecipitation, dominant-negative TSG101 overexpression, functional rescue assay, confocal microscopy\",\n      \"journal\": \"Nature medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction assays with functional rescue, independently replicated\",\n      \"pmids\": [\"11726971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"TSG101 binds to the PTAP motif of HIV-1 Gag p6 through its N-terminal UBC-like domain. Mutation of Tyr-110 (in place of the active-site Cys present in canonical E2 enzymes) and surrounding residues unique to TSG101 impairs p6 binding, showing that TSG101 is a catalytically inactive E2 variant whose distinctive features mediate viral protein interaction.\",\n      \"method\": \"Yeast two-hybrid, in vitro co-immunoprecipitation with purified proteins, in vivo co-immunoprecipitation in COS cells, site-directed mutagenesis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution with purified proteins + mutagenesis + multiple binding assays in one study\",\n      \"pmids\": [\"11427703\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"TSG101 (mammalian VPS23) functions in late endosomal trafficking. TSG101 mutant cells show defects in cathepsin D sorting/maturation, mislocalization of mannose-6-phosphate receptor, and recycling of endocytosed EGF receptors back to the cell surface instead of lysosomal delivery, demonstrating TSG101 is required for cargo delivery to late endosomes/MVBs.\",\n      \"method\": \"Cell fractionation, Western blotting, receptor trafficking assays in TSG101 mutant cells, pulse-chase\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal trafficking assays in TSG101-deficient cells, single lab\",\n      \"pmids\": [\"11208108\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The crystal/NMR structure of the TSG101 UEV domain reveals it resembles canonical E2 ubiquitin-conjugating enzymes but has an additional N-terminal helix, an extended beta-hairpin, and lacks two C-terminal helices. PTAP peptides bind in a hydrophobic cleft exposed by the absence of C-terminal helices; ubiquitin binds at a novel site surrounding the beta-hairpin. These two binding sites are spatially distinct and independent.\",\n      \"method\": \"NMR structure determination, chemical shift mapping, surface plasmon resonance binding assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — NMR structure with functional binding site mapping using orthogonal biochemical methods\",\n      \"pmids\": [\"12006492\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Solution structure of the TSG101 UEV domain in complex with the HIV-1 p6 PTAP peptide shows that each PTAP residue makes important contacts; the Ala-Pro dipeptide binds in a deep pocket resembling the X-Pro binding pockets of SH3 and WW domains, located above the vestigial active site.\",\n      \"method\": \"NMR solution structure determination\",\n      \"journal\": \"Nature structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — atomic-resolution NMR structure of the complex\",\n      \"pmids\": [\"12379843\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Crystal structure of the TSG101 UEV domain in complex with ubiquitin at 2.0 Å resolution shows TSG101 UEV contacts the Ile44 surface and an adjacent loop of ubiquitin through a highly solvated interface. Mutations disrupting this interface inhibit MVB sorting, and the ubiquitin-binding site is distinct from the PTAP-binding site, allowing independent binding of both ligands.\",\n      \"method\": \"X-ray crystallography, site-directed mutagenesis, functional MVB sorting assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with mutagenesis and functional validation\",\n      \"pmids\": [\"15053872\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"TSG101 directly interacts with mammalian VPS28 (hVPS28) through the conserved C-terminal portion of TSG101 to form part of a multiprotein complex (~350 kDa). Upon expression of dominant-negative VPS4, a portion of TSG101 and hVPS28 translocate from cytosol to the surface of aberrant endosomal vacuoles.\",\n      \"method\": \"Co-immunoprecipitation, chemical cross-linking, gel filtration, confocal microscopy with dominant-negative VPS4\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct interaction demonstrated by co-IP and cross-linking with domain mapping, functional localization experiment\",\n      \"pmids\": [\"11134028\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"TSG101 interacts with HRS (hepatocyte growth factor-regulated tyrosine kinase substrate) via the UEV domain of TSG101 binding to two proline-rich regions of HRS including a PSAP motif. Disruption of this interaction prevents delivery of EGFR to late endosomes, causes accumulation of ubiquitinated EGFR in early endosomes, and inhibits ligand-induced EGFR down-regulation.\",\n      \"method\": \"Co-immunoprecipitation, deletion/mutagenesis analysis, confocal microscopy, receptor trafficking assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with domain mapping plus functional consequence (receptor trafficking), single lab with multiple orthogonal methods\",\n      \"pmids\": [\"12802020\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"HIV-1 Gag mimics the Tsg101-recruiting activity of the endosomal protein Hrs. The Tsg101 UEV domain binds the PSAP motif (residues 348-351) within Hrs, and Hrs residues 222-777 can recruit Tsg101 and rescue budding of Gag particles lacking native late domains, demonstrating that Hrs normally recruits Tsg101 to the endosomal membrane and HIV Gag usurps this activity.\",\n      \"method\": \"Co-immunoprecipitation, yeast two-hybrid, functional rescue assay, dominant-negative competition\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding assays plus functional rescue, independently supported by parallel paper (PMID 12802020)\",\n      \"pmids\": [\"12900394\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"TSG101 participates in an autoregulatory loop with MDM2: the UBC domain of TSG101 interferes with ubiquitination of MDM2, stabilizing MDM2 and thereby down-regulating p53 protein levels. Conversely, elevated MDM2 promotes TSG101 degradation via the 26S proteasome.\",\n      \"method\": \"Pulse-chase analysis, Western blotting in wild-type and mutant fibroblasts, ubiquitination assays, MDM2/p53 overexpression experiments\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple biochemical methods in one lab establishing the regulatory loop, but no in vitro reconstitution of the direct inhibition of MDM2 ubiquitination\",\n      \"pmids\": [\"11172000\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"TSG101 null embryos accumulate p53 protein (without change in p53 mRNA), and introduction of a p53 null mutation into tsg101-/- embryos rescues the gastrulation defect and extends survival, establishing a functional genetic link between TSG101 and the p53 pathway in vivo.\",\n      \"method\": \"Gene targeting (knockout mice), genetic epistasis (double knockout), Western blotting, in vivo embryo analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis by double-knockout with defined phenotypic rescue, rigorous in vivo model\",\n      \"pmids\": [\"11172041\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Ubiquitylation of MHC class I by the Kaposi's sarcoma herpesvirus K3 protein requires TSG101 for subsequent degradation in the late endocytic pathway. siRNA depletion of TSG101 prevents MHC class I degradation and preserves its cell surface expression in K3-expressing cells.\",\n      \"method\": \"siRNA knockdown, flow cytometry, Western blotting, confocal microscopy\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean siRNA knockdown with defined functional readout, single lab\",\n      \"pmids\": [\"12006494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Tal (Tsg101-associated ligase) is an E3 ubiquitin ligase whose RING finger mediates multiple monoubiquitylation of TSG101. Bivalent binding of TSG101 to a tandem PTAP motif and a central region of Tal is required for Tal-mediated TSG101 ubiquitylation. Tal regulates a TSG101-associated complex responsible for sorting cargo into MVB vesicles and plasma membrane budding.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, deletion/mutagenesis analysis, functional assays (EGFR endocytosis, HIV egress)\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus domain mapping and functional validation, single lab\",\n      \"pmids\": [\"15256501\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Mahogunin E3 ubiquitin ligase interacts with the UEV domain of TSG101 via its PSAP motif and catalyzes monoubiquitylation of TSG101 both in vivo and in vitro. Depletion of Mahogunin disrupts endosome-to-lysosome trafficking of EGFR, resulting in prolonged downstream signaling.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, EGFR trafficking assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus interaction mapping and functional knockdown assay, single lab\",\n      \"pmids\": [\"17229889\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"The steadiness box (conserved C-terminal sequence) of TSG101 controls its steady-state level. Tal polyubiquitinates lysine residues in the C-terminus of TSG101 not complexed with other ESCRT-I proteins, leading to proteasomal degradation. VPS28 is a limiting factor, and ESCRT-I complex assembly protects TSG101 from Tal-mediated degradation.\",\n      \"method\": \"Ubiquitination assays, proteasome inhibitor experiments, siRNA knockdown, co-immunoprecipitation, Western blotting\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — in vitro and in vivo ubiquitination assays with domain mutagenesis and functional complementation, single lab\",\n      \"pmids\": [\"18077552\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"The TSG101 protein steady-state level is controlled posttranslationally by an evolutionarily conserved C-terminal sequence termed the 'steadiness box.' Overproduction of TSG101 from adventitious constructs results in compensatory down-regulation of endogenous TSG101 protein (not mRNA), through a proteolytic feedback loop involving the steadiness box.\",\n      \"method\": \"Western blotting, Northern blotting, deletion mapping of TSG101 constructs, pulse-chase in cultured cells\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple complementary experiments mapping the regulatory sequence, functional consequences assessed, single lab\",\n      \"pmids\": [\"10749147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"TSG101 localization is cell cycle-dependent: it localizes to the nucleus and Golgi complex during interphase, and to mitotic spindles and centrosomes during mitosis. TSG101-deficient cells display multiple microtubule organizing centers, aberrant mitotic spindles, aneuploidy, and nuclear anomalies.\",\n      \"method\": \"Indirect immunofluorescence, cell fractionation, FACS analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization by immunofluorescence across cell cycle stages with functional phenotyping of deficient cells, single lab\",\n      \"pmids\": [\"9465061\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Depletion of TSG101 inhibits EGF-stimulated MVB formation (number of MVBs per unit cytoplasm), causes tubulation of the early endosome vacuolar domains, and potently inhibits EGF degradation; this is distinct from the role of Hrs, which is required for accumulation of internal vesicles within MVBs but not MVB formation itself.\",\n      \"method\": \"siRNA depletion, electron microscopy, confocal microscopy, EGF degradation assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA with defined ultrastructural and functional readouts comparing TSG101 and Hrs roles, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"16707569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"In a cell-free reconstitution assay, the ESCRT-I subunit Tsg101 acts as a positive regulator of intraluminal vesicle (ILV) formation within late endosomes, while Alix acts as a negative regulator; ILV budding is inhibited by dominant-negative VPS4, confirming ESCRT dependence.\",\n      \"method\": \"In vitro budding reconstitution assay, dominant-negative mutant analysis, quantitative biochemistry\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of ILV budding with direct identification of Tsg101's positive regulatory role, single lab\",\n      \"pmids\": [\"18768755\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"TSG101 and ubiquitin are required for cSMAC formation at the immunological synapse. siRNA knockdown of TSG101 in primary T cells impairs cSMAC formation, TCR microcluster signal termination, TCR downregulation, and segregation of TCR-MHC-peptide from PKC-theta complexes, demonstrating that ubiquitin recognition by TSG101 is a molecular checkpoint for TCR downregulation.\",\n      \"method\": \"siRNA knockdown in primary T cells, live imaging of immunological synapses, TIRF microscopy, flow cytometry\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA in primary cells with multiple defined functional readouts, single lab with orthogonal imaging and signaling assays\",\n      \"pmids\": [\"20399684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TSG101 is recruited to the plasma membrane by binding the PSAP motif of the arrestin-domain protein ARRDC1, which drives direct plasma membrane budding to form ARRDC1-mediated microvesicles (ARMMs). This process requires VPS4 ATPase activity and is enhanced by the E3 ligase WWP2, which ubiquitinates ARRDC1.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis, live cell imaging, electron microscopy, siRNA knockdown\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with mutagenesis plus imaging and functional assays, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"22315426\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"TSG101 interacts with AATF and functions as a cooperative coactivator of androgen receptor-mediated transcription. TSG101 enhances monoubiquitination of the androgen receptor in a ligand-dependent manner, correlating with enhanced transactivating capacity.\",\n      \"method\": \"Co-immunoprecipitation, reporter gene assays, in vivo ubiquitination assay, dominant-negative ubiquitin overexpression\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with functional transcription assays and ubiquitination assays, single lab\",\n      \"pmids\": [\"14761944\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"TSG101 binds p21(Cip1/WAF1) and increases p21 protein stability in HEK293F cells and differentiating keratinocytes. In proliferating keratinocytes, TSG101 is recruited in a p21-dependent manner to cyclin/CDK complexes and inhibits cyclin/CDK activity, causing growth suppression.\",\n      \"method\": \"Co-immunoprecipitation, pulse-chase protein stability assays, kinase activity assays, antisense RNA knockdown, cell cycle analysis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP and functional kinase assays, multiple cell types, single lab\",\n      \"pmids\": [\"11943869\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"ALG-2 (a penta-EF-hand Ca2+-binding protein) directly binds the proline-rich region (PRR) of TSG101 in a Ca2+-dependent manner, as shown by GST pulldown and yeast two-hybrid. ALG-2 co-localizes with TSG101 at aberrant endosomes induced by dominant-negative SKD1/Vps4B, and this localization is Ca2+-dependent.\",\n      \"method\": \"GST pulldown, yeast two-hybrid, overlay assay with biotinylated ALG-2, immunofluorescence microscopy, deletion mapping\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — direct binding demonstrated by multiple in vitro assays with domain mapping, single lab\",\n      \"pmids\": [\"16004603\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"TSG101 stabilizes the unliganded (hypophosphorylated) form of the glucocorticoid receptor (GR) by impeding its proteasomal degradation and extending its half-life. The non-phosphorylated GR (S203A/S211A mutant) shows enhanced interaction with TSG101, and siRNA depletion of TSG101 renders hypophosphorylated GR unstable.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, pulse-chase analysis, proteasome inhibitor experiments, site-directed mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis plus functional stability assays, single lab\",\n      \"pmids\": [\"15657031\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"VPS13D directly interacts with TSG101 through its adaptor-binding domain, and together they remodel lipid droplet (LD) membranes to facilitate transfer of fatty acids from LDs to mitochondria at membrane contact sites. Depletion of TSG101 or VPS13D or ESCRT-III proteins inhibits fatty acid trafficking from LDs to mitochondria.\",\n      \"method\": \"Co-immunoprecipitation, in vitro LD membrane remodeling assay, lipid transfer binding assays, siRNA knockdown, live cell imaging\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of LD membrane remodeling plus multiple siRNA knockdowns with defined functional readouts, single lab\",\n      \"pmids\": [\"33623047\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TSG101 and Hrs are required for trafficking of ubiquitylated connexin-43 (Cx43) from early endosomes to lysosomes. siRNA depletion of Hrs or TSG101 individually abrogates Cx43 trafficking to lysosomes; simultaneous depletion causes accumulation of phosphorylated and ubiquitylated Cx43 in early endosomes.\",\n      \"method\": \"siRNA knockdown, confocal microscopy, Western blotting, protein half-life measurement, microinjected Lucifer yellow transfer assays\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA with multiple orthogonal functional readouts, single lab\",\n      \"pmids\": [\"19808888\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"ALG-2 bridges Alix and TSG101 as a Ca2+-dependent adaptor. ALG-2 is required for Ca2+-dependent pulldown of TSG101 by Strep-Alix; this bridge requires the ALG-2 binding site of Alix but not the PSAP motif; ALG-2 dimerization and Ca2+ binding are both required for bridging activity.\",\n      \"method\": \"Pulldown assays with purified proteins, ALG-2 knockdown, exogenous ALG-2 rescue, mutagenesis of ALG-2\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reconstitution with purified protein plus knockdown and rescue, single lab\",\n      \"pmids\": [\"19520058\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"TSG101 (ESCRT-I subunit) is required for nucleocapsid release from within multivesicular endosomes to the cytoplasm during vesicular stomatitis virus (VSV) infection, specifically by controlling the back-fusion of intra-endosomal vesicles with the endosome limiting membrane, independently of its role in viral envelope fusion.\",\n      \"method\": \"siRNA knockdown, infection assays, endosomal fractionation, functional infectivity measurements\",\n      \"journal\": \"Traffic (Copenhagen, Denmark)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA with defined functional consequence and mechanistic distinction from ESCRT-0 role, single lab\",\n      \"pmids\": [\"18817529\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TSG101 interacts with SQSTM1/p62 through its PRR domain and promotes p62 aggregation, leading to recruitment of Keap1 for autophagic degradation and release of Nrf2 to the nucleus. Knockout of p62 abrogates TSG101-induced cardioprotective effects, establishing TSG101 as a regulator of the p62/Keap1/Nrf2 signaling cascade.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion analysis, p62 knockout mice, transgenic and knockdown mouse models, RNA sequencing, pharmacological Nrf2 inhibition\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain mapping plus genetic epistasis in vivo, single lab\",\n      \"pmids\": [\"32057709\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TSG101 negatively regulates mitochondrial biogenesis in axons through a non-canonical, ESCRT-independent mechanism. Loss of Tsg101 activates PGC-1α/Nrf2-dependent mitochondrial biogenesis in an mTOR-independent, TFEB-dependent manner that requires the mitochondrial fission-fusion machinery. Loss of other ESCRT components does not recapitulate this phenotype.\",\n      \"method\": \"In vivo forward genetic screen (Drosophila), genetic epistasis with other ESCRT components, mitophagy and autophagy blockade, live imaging of axonal mitochondria\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo genetic screen with extensive epistasis analysis distinguishing ESCRT-independent mechanism, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"33972422\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TSG101 binds to PARP1 and is required for PARP1 activation and poly(ADP-ribose) (PAR) formation following DNA double-strand breaks. This TSG101 function is independent of its role in ESCRT-I. Absence of TSG101 impairs PAR-dependent formation of a nuclear PARP1-IKKγ signalosome, blocks IKK-NF-κB activation, causes PARP1 trapping at damage sites, and renders BRCA1/2-deficient cells synthetically lethal.\",\n      \"method\": \"Genome-wide siRNA screen, co-immunoprecipitation, PAR detection assays, NF-κB reporter assays, DNA repair assays, synthetic lethality assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — co-IP establishing direct binding plus multiple functional assays demonstrating ESCRT-independent role in DNA damage response, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"36124865\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TSG101 interacts with the coiled-coil domain of GISP (G protein-coupled receptor interacting scaffold protein) and mediates lysosomal degradation of GABA(B2) receptor. GISP overexpression inhibits TSG101-dependent GABA(B2) down-regulation, and a GISP mutant lacking the TSG101-binding domain has no protective effect.\",\n      \"method\": \"Co-immunoprecipitation from rat brain, GST pulldown, deletion mapping, siRNA/overexpression in HEK293 cells, receptor degradation assays\",\n      \"journal\": \"Journal of neurochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP from native tissue plus functional receptor stability assays, single lab\",\n      \"pmids\": [\"18643869\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TSG101 interacts with multiple connexins (Cx31, Cx43, Cx45 confirmed by co-immunoprecipitation from embryonic stem cells). siRNA-mediated knockdown of TSG101 increases levels of Cx43 and Cx45, prolongs their half-life, and increases gap junction-mediated dye transfer.\",\n      \"method\": \"Yeast two-hybrid, GST pulldown, co-immunoprecipitation from HM1 ES cells, siRNA knockdown, Western blotting, Lucifer yellow transfer\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — native co-IP plus functional connexin stability assays after siRNA, single lab\",\n      \"pmids\": [\"19210987\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"TSG101 interacts with the class II Rab11-FIPs (FIP3 and FIP4) through coiled-coil domains on both proteins. Point mutations in the coiled-coil regions of either TSG101 or FIP4 abrogate the interaction. Expression of TSG101 and FIP4 coiled-coil mutants causes cytokinesis defects, though the TSG101-FIP4 interaction is not required for TSG101 localization to the midbody during abscission.\",\n      \"method\": \"Proteomic screening, co-immunoprecipitation, site-directed mutagenesis, cytokinesis phenotype analysis, confocal microscopy\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP with mutagenesis plus functional cytokinesis phenotype, single lab\",\n      \"pmids\": [\"22348143\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Mahogunin (MGRN1) ubiquitinates TSG101, and this monoubiquitination is required for fusion of lysosomes with autophagosomes (via amphisomes) and late endosomes. Catalytically inactive MGRN1 or depletion of MGRN1 blocks these fusion events. Overexpression of TSG101 or its monoubiquitinated form rescues lysosomal fusion in MGRN1-deficient cells.\",\n      \"method\": \"In vivo and in vitro ubiquitination assays, siRNA knockdown, rescue by TSG101 overexpression, autophagy flux assays, lysosomal fusion assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional rescue experiments establishing the mechanistic link between TSG101 monoubiquitination and lysosomal fusion, single lab\",\n      \"pmids\": [\"26539917\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ALIX proline-rich domain (PRD) contains three tandem proline-rich motifs that compete for a single binding site on TSG101-UEV, as demonstrated by NMR relaxation dispersion and global fitting. This competitive interaction modulates ALIX function in ESCRT-mediated membrane remodeling.\",\n      \"method\": \"NMR spectroscopy (heteronuclear, relaxation dispersion), quantitative binding analysis by global fitting\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — atomic-resolution NMR with quantitative binding analysis, single lab but rigorous structural method\",\n      \"pmids\": [\"32917811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"TSG101 acts as a transcriptional suppressor through its coiled-coil domain. TSG101 represses estrogen receptor-mediated transcription and inhibits basal promoter activity; the coiled-coil domain is required for this repression and also for tumor suppressive function.\",\n      \"method\": \"Reporter gene transcription assays, deletion mutagenesis, histone acetyltransferase/deacetylase activity assays (negative result for these activities)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional transcription assays with domain mutagenesis, single lab\",\n      \"pmids\": [\"9588212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TSG101 interacts with HBV capsid protein HBc via the VFND motif in TSG101 and Lys-96 ubiquitination in HBc. NEDD4 (E3) and UbcH6 (E2) catalyze HBc ubiquitination required for TSG101-HBc interaction. TSG101 or NEDD4 knockdown reduces HBV particle counts in MVBs and suppresses HBV release.\",\n      \"method\": \"Co-immunoprecipitation, site-directed mutagenesis, in vitro ubiquitination assay, siRNA knockdown, transmission electron microscopy, HBV transgenic mouse model\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus site mutagenesis and functional validation in cells and in vivo, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"37224147\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TSG101 is a catalytically inactive ubiquitin E2 variant (UEV) that serves as the ubiquitin/PTAP-cargo-recognition subunit of the ESCRT-I complex: its UEV domain binds ubiquitylated cargo and PTAP/PSAP motifs in sorting adaptors (Hrs/HRS) and viral late domains to drive MVB/ILV biogenesis, receptor downregulation, and viral egress; it also functions independently of ESCRT-I in the nucleus (TSG101 is required for PARP1 activation and NF-κB signaling after DNA damage), in axons (suppressing PGC-1α/Nrf2-driven mitochondrial biogenesis), and at lipid droplet–mitochondria contacts (cooperating with VPS13D for fatty acid transfer), with its own steady-state levels controlled by the Tal/MGRN1-dependent monoubiquitination and proteasomal feedback loop acting through its C-terminal 'steadiness box.'\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TSG101 is the ubiquitin/peptide-cargo-recognition subunit of the ESCRT-I machinery that drives multivesicular body (MVB) biogenesis, receptor downregulation, and membrane scission [#3, #18]. Its catalytically inactive ubiquitin E2 variant (UEV) domain — structurally an E2-fold lacking the active-site cysteine and C-terminal helices — engages two spatially distinct ligands at independent sites: monoubiquitin via the Ile44 surface and PTAP/PSAP tetrapeptide motifs in a hydrophobic SH3/WW-like pocket [#4, #5, #6]. Through these surfaces TSG101 recognizes ubiquitylated cargo and is recruited to endosomal membranes by the PSAP motif of the ESCRT-0 adaptor HRS, then assembles with VPS28 into the ~350 kDa ESCRT-I complex to deliver cargo such as EGFR and connexins to late endosomes/lysosomes in a VPS4-dependent manner [#7, #8, #9, #27]. This same cargo-recognition activity is usurped by enveloped viruses: the PTAP late domain of HIV-1 Gag and PTAP-like motifs of Ebola VP40 and HBV capsid directly bind the UEV domain to recruit ESCRT-I and complete budding [#0, #1, #2, #39]. TSG101 likewise mediates plasma-membrane ARMM budding via ARRDC1 and acts as a positive regulator of intraluminal vesicle formation, antagonized by ALIX, which competes for the UEV peptide-binding site and is bridged to TSG101 by Ca2+-loaded ALG-2 [#19, #21, #37, #28]. Beyond canonical sorting, TSG101 performs ESCRT-independent functions: in the nucleus it binds PARP1 and is required for PARP1 activation, NF-\\u03baB signaling after DNA damage, and synthetic lethality in BRCA1/2-deficient cells [#32]; in axons it suppresses PGC-1\\u03b1/Nrf2-driven mitochondrial biogenesis [#31]; and at lipid droplet\\u2013mitochondria contacts it cooperates with VPS13D to remodel membranes for fatty-acid transfer [#26]. Its own abundance is set by a C-terminal 'steadiness box' through which the E3 ligases Tal and MGRN1 monoubiquitinate TSG101, with non-ESCRT-incorporated TSG101 polyubiquitinated for proteasomal degradation unless protected by VPS28/ESCRT-I assembly [#13, #14, #15]. TSG101 loss in vivo stabilizes p53, and a p53-null background rescues the gastrulation defect of TSG101-null embryos, linking it genetically to the p53 pathway [#11].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Established TSG101's cell-cycle-dependent subcellular distribution and a role in genome stability, raising the question of its biochemical activity.\",\n      \"evidence\": \"Immunofluorescence across the cell cycle and phenotyping of deficient cells\",\n      \"pmids\": [\"9465061\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular mechanism for spindle/centrosome function\", \"Relationship to later ESCRT role unresolved\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Identified TSG101 as a VPS23 ortholog acting in late-endosomal cargo delivery and showed its abundance is set posttranslationally by a C-terminal steadiness box, defining its trafficking role and an autoregulatory layer.\",\n      \"evidence\": \"Trafficking assays in mutant cells; deletion mapping and pulse-chase; co-IP/cross-linking with VPS28\",\n      \"pmids\": [\"11208108\", \"10749147\", \"11134028\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Steadiness-box ligase not yet identified\", \"Mechanism of cargo selection not yet structural\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Defined the UEV domain as a catalytically inactive E2 variant that binds ubiquitin and PTAP late domains, establishing TSG101/ESCRT-I as the cellular machinery hijacked for HIV-1 and Ebola budding.\",\n      \"evidence\": \"siRNA depletion with rescue, binding assays, dominant-negative VPS4, EM; site-directed mutagenesis of Tyr-110\",\n      \"pmids\": [\"11595185\", \"11726971\", \"11427703\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Atomic basis of dual-ligand recognition not yet solved\", \"Composition of full ESCRT-I in budding incomplete\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Linked TSG101 to the p53/MDM2 axis in vivo, showing its loss stabilizes p53 and that p53 ablation rescues the null embryonic phenotype.\",\n      \"evidence\": \"Knockout and double-knockout mice with genetic epistasis; MDM2/p53 biochemistry\",\n      \"pmids\": [\"11172041\", \"11172000\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct mechanism of MDM2 ubiquitination interference not reconstituted\", \"Whether this is ESCRT-dependent unclear\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Solved UEV-domain structures alone, with PTAP peptide, establishing distinct, independent binding pockets for peptide ligands and the vestigial active site.\",\n      \"evidence\": \"NMR structures with SPR/chemical-shift mapping\",\n      \"pmids\": [\"12006492\", \"12379843\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitin-binding geometry not yet co-crystallized (resolved later)\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Identified HRS as the physiological PSAP adaptor recruiting TSG101 to endosomes, explaining how cellular cargo sorting and viral budding share the same recruitment logic.\",\n      \"evidence\": \"Reciprocal co-IP, yeast two-hybrid, functional rescue and EGFR trafficking assays\",\n      \"pmids\": [\"12802020\", \"12900394\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of HRS-TSG101 handoff unclear\", \"How ubiquitin and PSAP binding are coordinated unresolved\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Determined the UEV-ubiquitin co-crystal structure and identified Tal as an E3 ligase that monoubiquitinates TSG101, defining both the ubiquitin-recognition interface and a regulatory ubiquitination input.\",\n      \"evidence\": \"X-ray crystallography with mutagenesis/MVB assays; in vitro ubiquitination and domain mapping\",\n      \"pmids\": [\"15053872\", \"15256501\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of monoubiquitination not fully defined here\", \"Whether Tal acts on assembled vs free TSG101 unclear\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Distinguished TSG101's role in MVB formation itself from HRS's role in ILV accumulation, separating ESCRT-I and ESCRT-0 functions.\",\n      \"evidence\": \"siRNA with EM and EGF-degradation assays\",\n      \"pmids\": [\"16707569\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Biophysical mechanism of membrane deformation not addressed\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Showed MGRN1 (Mahogunin) monoubiquitinates TSG101 and that the steadiness box governs Tal-mediated polyubiquitination/degradation gated by VPS28 availability, defining how ESCRT-I assembly protects TSG101.\",\n      \"evidence\": \"In vitro ubiquitination, proteasome inhibition, siRNA, co-IP\",\n      \"pmids\": [\"17229889\", \"18077552\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Switch between mono- and poly-ubiquitination not fully resolved\", \"In vivo relevance of feedback loop untested\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Reconstituted ILV budding in vitro and assigned TSG101 a positive regulatory role antagonized by ALIX; extended ESCRT-I function to viral nucleocapsid back-fusion.\",\n      \"evidence\": \"Cell-free budding reconstitution; VSV infection and endosomal fractionation\",\n      \"pmids\": [\"18768755\", \"18817529\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular basis of ALIX antagonism not yet structural (resolved 2020)\", \"Back-fusion mechanism single-lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Generalized TSG101 recruitment beyond endosomes to direct plasma-membrane ARMM budding via ARRDC1 and to midbody/cytokinesis via Rab11-FIP coiled-coil interactions.\",\n      \"evidence\": \"Co-IP, mutagenesis, live imaging, EM; cytokinesis phenotyping\",\n      \"pmids\": [\"22315426\", \"22348143\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"FIP4 interaction dispensable for midbody localization, leaving recruitment mechanism open\", \"ARMM cargo selectivity unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Resolved how ALIX competes with itself and TSG101 at the UEV peptide site and uncovered a p62/Keap1/Nrf2 cardioprotective signaling role, broadening TSG101 function.\",\n      \"evidence\": \"NMR relaxation dispersion; co-IP, domain mapping, p62-knockout mice\",\n      \"pmids\": [\"32917811\", \"32057709\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological balance of ALIX vs cargo competition in cells unmeasured\", \"Direct vs indirect p62 effects not fully separated\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined two ESCRT-independent functions: lipid-droplet-to-mitochondria fatty-acid transfer with VPS13D, and suppression of axonal mitochondrial biogenesis via PGC-1\\u03b1/Nrf2.\",\n      \"evidence\": \"In vitro LD remodeling and lipid transfer; Drosophila forward genetic screen with ESCRT epistasis\",\n      \"pmids\": [\"33623047\", \"33972422\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a single protein partitions between ESCRT and non-ESCRT pools unknown\", \"Direct biochemical activity in mitochondrial biogenesis suppression undefined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established a nuclear, ESCRT-independent role in which TSG101 binds and activates PARP1 to drive NF-\\u03baB signaling and confers synthetic lethality in BRCA1/2-deficient cells.\",\n      \"evidence\": \"Genome-wide siRNA screen, co-IP, PAR and NF-\\u03baB assays, synthetic-lethality assays\",\n      \"pmids\": [\"36124865\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How TSG101 mechanistically promotes PARP1 catalysis unresolved\", \"Nuclear targeting signal/regulation not defined\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended viral-egress recruitment to HBV, requiring NEDD4/UbcH6-catalyzed ubiquitination of the HBc capsid and a VFND motif in TSG101.\",\n      \"evidence\": \"In vitro ubiquitination, mutagenesis, siRNA, EM, transgenic mouse\",\n      \"pmids\": [\"37224147\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of VFND-mediated contact undefined\", \"Whether full ESCRT-I is required for HBV release untested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How TSG101 is partitioned and regulated between ESCRT-I-bound cytoplasmic pools and its ESCRT-independent nuclear, axonal, and membrane-contact-site roles remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mechanism for nuclear import/retention\", \"No structure of full ESCRT-I human complex in the timeline\", \"Determinants directing TSG101 to non-ESCRT functions unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0031386\", \"supporting_discovery_ids\": [0, 4, 6]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [8, 9, 21]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [10, 25, 32]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [26]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [3, 7, 18]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [21]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [17, 32]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [17]},\n      {\"term_id\": \"GO:0005811\", \"supporting_discovery_ids\": [26]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [3, 8, 18, 19]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [0, 1, 39]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [32]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [30, 36]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [13, 14, 15]}\n    ],\n    \"complexes\": [\"ESCRT-I\"],\n    \"partners\": [\"VPS28\", \"HRS\", \"ALIX\", \"ALG-2\", \"ARRDC1\", \"VPS13D\", \"PARP1\", \"MGRN1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}