{"gene":"TSPYL5","run_date":"2026-06-10T10:51:56","timeline":{"discoveries":[{"year":2010,"finding":"TSPYL5 physically interacts with USP7 (ubiquitin-specific protease 7), the deubiquitylase for p53, and reduces USP7 deubiquitylase activity toward p53, resulting in increased p53 ubiquitylation and decreased p53 protein levels. TSPYL5 thereby inhibits p53-target gene activation and overrides p53-dependent proliferation arrest and oncogene-induced senescence.","method":"Mass spectrometric analysis of TSPYL5-interacting proteins, co-immunoprecipitation, ubiquitylation assays, cell-based proliferation and senescence assays","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — mass spectrometry identification of binding partner, functional assays (ubiquitylation, proliferation arrest, senescence), multiple orthogonal methods, widely replicated by subsequent studies","pmids":["21170034"],"is_preprint":false},{"year":2019,"finding":"TSPYL5 co-localizes with ALT telomeres in PML bodies and prevents poly-ubiquitination and proteasomal degradation of POT1 (a shelterin component) exclusively in ALT+ cancer cells. USP7 depletion rescued POT1 poly-ubiquitination and loss, indicating that USP7 activates POT1 E3 ubiquitin ligase(s); PML depletion also suppressed POT1 poly-ubiquitination, suggesting interplay between USP7 and PML in triggering POT1 degradation when TSPYL5 is absent.","method":"TSPYL5 depletion (siRNA/shRNA), co-localization studies, ubiquitination assays, proteasome inhibitor experiments, USP7 and PML knockdown epistasis experiments","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal epistasis experiments (USP7 KD and PML KD rescuing POT1 loss), multiple orthogonal methods, published in high-impact journal","pmids":["31278054"],"is_preprint":false},{"year":2010,"finding":"TSPYL5 suppression in A549 lung cancer cells up-regulates PTEN, leading to down-regulation of AKT activation, and also increases p21(WAF1/Cip1) levels, causing cell growth inhibition and sensitization to gamma-radiation. Overexpression of TSPYL5 in H460 cells produced the opposite effects.","method":"siRNA knockdown, TSPYL5 overexpression, Western blotting for PTEN/AKT/p21, colony formation assay, gamma-irradiation survival assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — loss-of-function and gain-of-function with defined molecular readouts in two cell lines, single lab","pmids":["20079711"],"is_preprint":false},{"year":2013,"finding":"TSPYL5 directly binds to the CYP19A1 (aromatase) promoter I.4 and functions as a transcriptional activator of CYP19A1 expression; TSPYL5 knockdown decreased and overexpression increased aromatase expression in MCF-7 cells, lymphoblastoid cell lines, and adipocytes. A putative TSPYL5 binding motif was identified in 43 genes, suggesting broader transcription factor activity.","method":"Chromatin immunoprecipitation (ChIP), TSPYL5 knockdown and overexpression in multiple cell types, reporter assays identifying functional estrogen response element at rs2583506","journal":"Molecular endocrinology (Baltimore, Md.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP assay directly demonstrates promoter binding, functional validation across multiple cell types, single lab","pmids":["23518928"],"is_preprint":false},{"year":2017,"finding":"MUC16 regulates TSPYL5 expression through the JAK2/STAT3/glucocorticoid receptor (GR) axis in lung cancer cells; MUC16 knockdown decreased TSPYL5, and inhibition of STAT3 (Y705) led to decreased GR and TSPYL5, while MUC16-Cter overexpression rescued these signaling proteins and TSPYL5 expression.","method":"Stable shRNA knockdown of MUC16, transcriptome analysis, MUC16-Cter overexpression rescue, STAT3 inhibitor treatment, Western blotting","journal":"Clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — epistasis established via rescue and inhibitor experiments, transcriptomic identification, single lab","pmids":["28196872"],"is_preprint":false},{"year":2018,"finding":"TSPYL5 promotes human endothelial cell (EC) proliferation, migration, and tube formation by downregulating p53 expression; TSPYL5 depletion-mediated loss of EC functions was rescued by p53 inhibition, and TSPYL5 overexpression blocked adriamycin-induced senescence. In vivo, TSPYL5 overexpression promoted angiogenesis and wound healing.","method":"TSPYL5 overexpression and knockdown in HUVECs and hPSC-derived ECs, p53 inhibitor rescue, Matrigel plug angiogenesis assay, mouse wound healing model","journal":"Angiogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (p53 inhibitor rescue of TSPYL5-KD phenotype), in vivo validation, single lab","pmids":["30471052"],"is_preprint":false},{"year":2021,"finding":"AKT phosphorylates TSPYL5 at threonine-120, stabilizing it by inhibiting its ubiquitination. Phosphorylated TSPYL5-pT120 translocates to the nucleus and acts as a transcriptional activator of CSC-associated genes ALDH1 and CD44, while also suppressing PTEN transcription, creating a positive feedback loop (AKT/TSPYL5/PTEN) that maintains cancer stem cell characteristics.","method":"Phosphorylation site mutagenesis (T120 substitution), ubiquitination assays, nuclear fractionation, transcriptional reporter assays, ChIP, TSPYL5 knockdown and overexpression in NSCLC cells","journal":"Communications biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — mutagenesis of phosphorylation site combined with ubiquitination assays, nuclear fractionation, transcriptional assays, and in vivo validation; multiple orthogonal methods","pmids":["34163000"],"is_preprint":false},{"year":2023,"finding":"TSPYL5 localizes to both the cytoplasm and nucleoplasm (including the nucleolus); its nuclear/nucleolar localization is mediated by NLS/NoLS sequences in the N-terminal intrinsically disordered region (residues 4–27), while cytoplasmic localization is regulated by the ordered NAP-like domain (residues 198–382). The N-terminal disordered region (1–198 aa) contributes to exchange dynamics with the nucleoplasm and potential phase separation.","method":"Live-cell imaging of TSPYL5-EGFP fusion proteins, FRAP, deletion/truncation constructs, bioinformatics analysis of interactome","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct live imaging and FRAP with functional domain constructs, single lab, no full reconstitution","pmids":["38203210"],"is_preprint":false},{"year":2024,"finding":"Mouse Tspyl5 promotes spermatogonia proliferation by facilitating ubiquitination-mediated degradation of TRP53 (p53), which upregulates Pcna expression and enhances PCNA-mediated DNA replication. Tspyl5 knockout in aged mice caused reduced spermatogonia numbers and spermatozoa, and transcriptomics revealed downregulation of the Pcna-mediated DNA replication pathway.","method":"Tspyl5 knockout mouse model, transcriptomic analysis of spermatogonia, ubiquitination assays for TRP53 degradation, Western blotting for PCNA","journal":"Reproduction, fertility, and development","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo knockout with defined molecular readout (p53 ubiquitination and PCNA), single lab, mouse model","pmids":["38185096"],"is_preprint":false},{"year":2025,"finding":"TSPYL5 binds G3BP1 and enhances G3BP1 Ser149 phosphorylation, driving G3BP1 nuclear membrane translocation. This recruits p53 for nucleoporin RanBP2, forming a RanBP2-G3BP1-p53 complex that accelerates p53 sumoylation and nuclear export, sequestering p53 in the cytoplasm and suppressing its transcription factor function in neuroblastoma cells.","method":"Co-immunoprecipitation, phosphorylation assays, sumoylation assays, nuclear fractionation, complex reconstitution experiments, TSPYL5 overexpression and depletion in neuroblastoma cells","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP and mechanistic epistasis with multiple binding partners identified, single lab, single study","pmids":["40319028"],"is_preprint":false},{"year":2026,"finding":"TSPYL5 sequesters the deubiquitinase USP10, preventing USP10 from stabilizing PTEN, and thereby triggering proteasomal degradation of PTEN in triple-negative breast cancer (TNBC) cells. This TSPYL5-USP10-PTEN axis hyperactivates PI3K/AKT signaling and unleashes a ZEB1-driven EMT/metastatic program, even in cells retaining wild-type PTEN.","method":"Co-immunoprecipitation (TSPYL5-USP10 interaction), ubiquitination assays, PTEN stability assays, TSPYL5 overexpression driving orthotopic tumor polymetastasis in animals, single-cell and spatial transcriptomic analyses","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP identifying novel binding partner USP10, functional ubiquitination/stability assays, in vivo metastasis model; single lab, single study","pmids":["42241080"],"is_preprint":false},{"year":2020,"finding":"TSPYL5 overexpression in colorectal cancer cells activates endoplasmic reticulum stress (ERS), suppresses cell proliferation, migration, and invasion, and induces apoptosis; upregulation of caspase-1, caspase-3, Bax, ATF4, and CHOP was observed after TSPYL5 overexpression.","method":"Transfection of pcDNA3.1-TSPYL5 in HCT116 and HT29 cells, EdU proliferation assay, flow cytometry, Transwell assay, transmission electron microscopy for ERS, Western blotting","journal":"Oncology reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — overexpression with cellular phenotype readout and ERS markers, but no defined upstream molecular mechanism; single lab, no pathway placement beyond ERS induction","pmids":["32627024"],"is_preprint":false}],"current_model":"TSPYL5 is an intrinsically disordered nucleosome assembly protein (NAP) family member that suppresses p53 function through multiple mechanisms: it inhibits USP7 deubiquitylase activity toward p53 (increasing p53 ubiquitylation and degradation), drives p53 nuclear export via a G3BP1-RanBP2 sumoylation axis, and promotes p53 ubiquitination in spermatogonia to upregulate PCNA-mediated DNA replication; additionally, TSPYL5 protects POT1 from proteasomal degradation in ALT+ cancer cells by antagonizing USP7-dependent ubiquitination, is phosphorylated by AKT at T120 to translocate to the nucleus where it transcriptionally activates CSC genes and suppresses PTEN, sequesters USP10 to trigger PTEN proteasomal degradation in TNBC, and functions as a transcriptional activator of CYP19A1 (aromatase) by directly binding its promoter."},"narrative":{"mechanistic_narrative":"TSPYL5 is a nucleosome assembly protein (NAP)-family member that acts predominantly as a negative regulator of p53, governing proliferation, senescence, and stemness in normal and cancer cells [PMID:21170034, PMID:30471052]. Its founding biochemical activity is direct binding to the deubiquitylase USP7, which it inhibits to drive p53 ubiquitylation and degradation, thereby overriding p53-dependent proliferation arrest and oncogene-induced senescence [PMID:21170034]. TSPYL5 deploys this anti-p53 logic in multiple contexts: in endothelial cells it lowers p53 to promote proliferation, migration, and angiogenesis [PMID:30471052]; in spermatogonia it facilitates ubiquitination-mediated TRP53 degradation to upregulate PCNA-driven DNA replication [PMID:38185096]; and in neuroblastoma it binds G3BP1 and enhances its Ser149 phosphorylation, nucleating a RanBP2–G3BP1–p53 complex that accelerates p53 sumoylation and nuclear export to sequester p53 in the cytoplasm [PMID:40319028]. Beyond p53, TSPYL5 sustains PI3K/AKT signaling by suppressing PTEN — directly downregulating PTEN to lower AKT activity when depleted [PMID:20079711], and in triple-negative breast cancer sequestering the deubiquitinase USP10 to trigger PTEN proteasomal degradation and a ZEB1-driven EMT program [PMID:42241080]. AKT in turn phosphorylates TSPYL5 at Thr120, stabilizing it and driving its nuclear translocation where it transcriptionally activates the stem-cell genes ALDH1 and CD44 while repressing PTEN, forming a self-reinforcing AKT/TSPYL5/PTEN feedback loop [PMID:34163000]. TSPYL5 also functions as a sequence-specific transcriptional activator, binding the CYP19A1 (aromatase) promoter I.4 to upregulate aromatase expression [PMID:23518928]. In ALT+ cancer cells it localizes to telomeric PML bodies and protects the shelterin component POT1 from USP7- and PML-dependent ubiquitination and degradation [PMID:31278054]. TSPYL5 is an intrinsically disordered protein whose N-terminal disordered region (residues 4–27) carries NLS/NoLS signals directing nuclear/nucleolar localization, while its ordered NAP-like domain directs cytoplasmic localization [PMID:38203210].","teleology":[{"year":2010,"claim":"Established the founding molecular mechanism: how TSPYL5 controls cell fate by linking it to the p53 ubiquitylation machinery rather than acting through an unknown route.","evidence":"Mass spectrometry of TSPYL5 interactors, co-IP, ubiquitylation assays, and proliferation/senescence assays identifying USP7 inhibition and p53 destabilization","pmids":["21170034"],"confidence":"High","gaps":["Does not define the structural basis of USP7 inhibition","Does not establish whether TSPYL5 acts catalytically or stoichiometrically on USP7"]},{"year":2010,"claim":"Connected TSPYL5 to a second tumor-suppressor axis by showing it regulates the PTEN/AKT pathway and the radiation response, broadening its role beyond p53.","evidence":"siRNA knockdown and overexpression in lung cancer lines with PTEN/AKT/p21 Western blots, colony formation, and gamma-irradiation survival assays","pmids":["20079711"],"confidence":"Medium","gaps":["Mechanism by which TSPYL5 regulates PTEN not defined at this stage","Single lab, two cell lines"]},{"year":2013,"claim":"Demonstrated TSPYL5 has direct sequence-specific transcriptional activity, establishing it as a DNA-binding transactivator rather than solely a protein-interaction scaffold.","evidence":"ChIP showing binding to CYP19A1 promoter I.4, knockdown/overexpression across multiple cell types, and reporter assays","pmids":["23518928"],"confidence":"Medium","gaps":["No structural definition of the DNA-binding interface","The broader 43-gene putative target set remains uncharacterized functionally"]},{"year":2017,"claim":"Placed TSPYL5 downstream of an upstream signaling cascade, identifying how its expression is controlled in cancer.","evidence":"MUC16 shRNA knockdown, transcriptome analysis, MUC16-Cter rescue, and STAT3 inhibition in lung cancer cells","pmids":["28196872"],"confidence":"Medium","gaps":["Direct vs indirect transcriptional control of TSPYL5 not resolved","Single lab"]},{"year":2018,"claim":"Extended the anti-p53 function to a physiological vascular context, showing TSPYL5 promotes angiogenesis by suppressing p53.","evidence":"Overexpression/knockdown in HUVECs and hPSC-ECs with p53-inhibitor rescue, Matrigel plug, and mouse wound-healing models","pmids":["30471052"],"confidence":"Medium","gaps":["Does not establish whether the USP7 mechanism operates in endothelial cells","Single lab"]},{"year":2019,"claim":"Revealed a p53-independent role at telomeres, showing TSPYL5 protects POT1 from degradation specifically in ALT+ cancer cells.","evidence":"TSPYL5 depletion, PML-body co-localization, ubiquitination and proteasome-inhibitor assays, and USP7/PML knockdown epistasis","pmids":["31278054"],"confidence":"High","gaps":["Identity of the POT1 E3 ligase activated by USP7 not established","Why protection is ALT-specific not mechanistically resolved"]},{"year":2021,"claim":"Identified the post-translational switch governing TSPYL5 nuclear function, defining a feedback loop that sustains cancer stemness.","evidence":"T120 phosphosite mutagenesis, ubiquitination assays, nuclear fractionation, transcriptional reporters and ChIP in NSCLC cells","pmids":["34163000"],"confidence":"High","gaps":["Direct vs indirect PTEN promoter repression not fully separated from ALDH1/CD44 activation","Identity of the E3 ligase counteracted by T120 phosphorylation unknown"]},{"year":2023,"claim":"Mapped the determinants of TSPYL5 subcellular partitioning, linking its disordered N-terminus to nuclear/nucleolar targeting and its NAP domain to cytoplasmic retention.","evidence":"Live-cell imaging of EGFP fusions, FRAP, and deletion/truncation constructs","pmids":["38203210"],"confidence":"Medium","gaps":["Phase separation only inferred, not reconstituted","Functional consequence of nucleolar pool not defined"]},{"year":2024,"claim":"Provided in vivo genetic evidence for a physiological role, showing Tspyl5 drives spermatogonia proliferation via p53 degradation and PCNA upregulation.","evidence":"Tspyl5 knockout mice, spermatogonia transcriptomics, TRP53 ubiquitination assays, and PCNA Westerns","pmids":["38185096"],"confidence":"Medium","gaps":["Does not confirm the USP7 mechanism operates in spermatogonia","Single lab, mouse model"]},{"year":2025,"claim":"Defined a new p53-suppressing mechanism via spatial regulation, showing TSPYL5 drives p53 nuclear export and sumoylation through a G3BP1-RanBP2 axis.","evidence":"Co-IP, phosphorylation and sumoylation assays, nuclear fractionation, and complex reconstitution in neuroblastoma cells","pmids":["40319028"],"confidence":"Medium","gaps":["Direct kinase mediating G3BP1 Ser149 phosphorylation not identified","Single lab, single study"]},{"year":2026,"claim":"Identified USP10 as a new TSPYL5 partner explaining PTEN destabilization in TNBC even with wild-type PTEN, linking TSPYL5 to EMT and metastasis.","evidence":"Co-IP of TSPYL5-USP10, ubiquitination/stability assays, orthotopic metastasis models, and single-cell/spatial transcriptomics","pmids":["42241080"],"confidence":"Medium","gaps":["Reciprocal validation of the sequestration mechanism limited","Single lab, single study"]},{"year":null,"claim":"It remains unresolved how TSPYL5's distinct activities — USP7/USP10 deubiquitylase modulation, sequence-specific transcription, telomeric POT1 protection, and p53 spatial control — are coordinated as a single biochemical mechanism versus context-dependent moonlighting functions.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structure of TSPYL5 or its complexes","No unifying biochemical model linking its deubiquitylase-regulatory and transcriptional activities","Whether NAP-family nucleosome assembly activity is biochemically active is untested in the corpus"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[3,6]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[3]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,10]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[10]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[7]},{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[6,7]},{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[7]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[6]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,6,10]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[3,6]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,10]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,5]}],"complexes":["RanBP2-G3BP1-p53 complex"],"partners":["USP7","USP10","G3BP1","POT1","RANBP2","P53"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86VY4","full_name":"Testis-specific Y-encoded-like protein 5","aliases":[],"length_aa":417,"mass_kda":45.1,"function":"Involved in modulation of cell growth and cellular response to gamma radiation probably via regulation of the Akt signaling pathway. Involved in regulation of p53/TP53. Suppresses p53/TP53 protein levels and promotes its ubiquitination; the function is dependent on USP7 and independent on MDM2. Proposed to displace p53/TP53 from interaction with USP7","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/Q86VY4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/TSPYL5","classification":"Common Essential","n_dependent_lines":600,"n_total_lines":1208,"dependency_fraction":0.4966887417218543},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/TSPYL5","total_profiled":1310},"omim":[{"mim_id":"614721","title":"TSPY-LIKE 5; TSPYL5","url":"https://www.omim.org/entry/614721"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Golgi apparatus","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"testis","ntpm":41.8}],"url":"https://www.proteinatlas.org/search/TSPYL5"},"hgnc":{"alias_symbol":["KIAA1750"],"prev_symbol":[]},"alphafold":{"accession":"Q86VY4","domains":[{"cath_id":"3.30.1120.90","chopping":"250-386","consensus_level":"high","plddt":86.992,"start":250,"end":386},{"cath_id":"1.20.5","chopping":"208-248","consensus_level":"medium","plddt":92.4295,"start":208,"end":248}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86VY4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86VY4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86VY4-F1-predicted_aligned_error_v6.png","plddt_mean":61.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TSPYL5","jax_strain_url":"https://www.jax.org/strain/search?query=TSPYL5"},"sequence":{"accession":"Q86VY4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86VY4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86VY4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86VY4"}},"corpus_meta":[{"pmid":"21170034","id":"PMC_21170034","title":"TSPYL5 suppresses p53 levels and function by physical interaction with USP7.","date":"2010","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/21170034","citation_count":106,"is_preprint":false},{"pmid":"28196872","id":"PMC_28196872","title":"MUC16 Regulates TSPYL5 for Lung Cancer Cell Growth and Chemoresistance by Suppressing p53.","date":"2017","source":"Clinical cancer research : an official journal of the American Association for Cancer Research","url":"https://pubmed.ncbi.nlm.nih.gov/28196872","citation_count":89,"is_preprint":false},{"pmid":"31278054","id":"PMC_31278054","title":"TSPYL5 Depletion Induces Specific Death of ALT Cells through USP7-Dependent Proteasomal Degradation of POT1.","date":"2019","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/31278054","citation_count":47,"is_preprint":false},{"pmid":"23518928","id":"PMC_23518928","title":"TSPYL5 SNPs: association with plasma estradiol concentrations and aromatase expression.","date":"2013","source":"Molecular endocrinology (Baltimore, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/23518928","citation_count":45,"is_preprint":false},{"pmid":"18059362","id":"PMC_18059362","title":"Gene silencing of TSPYL5 mediated by aberrant promoter methylation in gastric cancers.","date":"2007","source":"Laboratory investigation; a journal of technical methods and pathology","url":"https://pubmed.ncbi.nlm.nih.gov/18059362","citation_count":40,"is_preprint":false},{"pmid":"20079711","id":"PMC_20079711","title":"TSPYL5 is involved in cell growth and the resistance to radiation in A549 cells via the regulation of p21(WAF1/Cip1) and PTEN/AKT pathway.","date":"2010","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/20079711","citation_count":37,"is_preprint":false},{"pmid":"26386860","id":"PMC_26386860","title":"Hypermethylation of ACP1, BMP4, and TSPYL5 in Hepatocellular Carcinoma and Their Potential Clinical Significance.","date":"2015","source":"Digestive diseases and sciences","url":"https://pubmed.ncbi.nlm.nih.gov/26386860","citation_count":34,"is_preprint":false},{"pmid":"28716051","id":"PMC_28716051","title":"miR-380-5p-mediated repression of TEP1 and TSPYL5 interferes with telomerase activity and favours the emergence of an \"ALT-like\" phenotype in diffuse malignant peritoneal mesothelioma cells.","date":"2017","source":"Journal of hematology & oncology","url":"https://pubmed.ncbi.nlm.nih.gov/28716051","citation_count":28,"is_preprint":false},{"pmid":"30471052","id":"PMC_30471052","title":"TSPYL5-mediated inhibition of p53 promotes human endothelial cell function.","date":"2018","source":"Angiogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/30471052","citation_count":25,"is_preprint":false},{"pmid":"34163000","id":"PMC_34163000","title":"Targeting therapy-resistant lung cancer stem cells via disruption of the AKT/TSPYL5/PTEN positive-feedback 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by Targeting TSPYL5.","date":"2017","source":"DNA and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/29240449","citation_count":11,"is_preprint":false},{"pmid":"32627024","id":"PMC_32627024","title":"TSPYL5 activates endoplasmic reticulum stress to inhibit cell proliferation, migration and invasion in colorectal cancer.","date":"2020","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/32627024","citation_count":10,"is_preprint":false},{"pmid":"38185096","id":"PMC_38185096","title":"Mouse Tspyl5 promotes spermatogonia proliferation through enhancing Pcna-mediated DNA replication.","date":"2024","source":"Reproduction, fertility, and development","url":"https://pubmed.ncbi.nlm.nih.gov/38185096","citation_count":3,"is_preprint":false},{"pmid":"38203210","id":"PMC_38203210","title":"Insights into the Cellular Localization and Functional Properties of TSPYL5 Protein.","date":"2023","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38203210","citation_count":2,"is_preprint":false},{"pmid":"37675714","id":"PMC_37675714","title":"TSPYL5 inhibits the tumorigenesis of colorectal cancer cells in vivo by triggering DNA damage.","date":"2023","source":"Journal of cancer research and therapeutics","url":"https://pubmed.ncbi.nlm.nih.gov/37675714","citation_count":2,"is_preprint":false},{"pmid":"31692925","id":"PMC_31692925","title":"Specific death of ALT cells through TSPYL5 depletion.","date":"2019","source":"Molecular & cellular oncology","url":"https://pubmed.ncbi.nlm.nih.gov/31692925","citation_count":2,"is_preprint":false},{"pmid":"40319028","id":"PMC_40319028","title":"TSPYL5-driven G3BP1 nuclear membrane translocation facilitates p53 cytoplasm sequestration via accelerating RanBP2-mediated p53 sumoylation and nuclear export in neuroblastoma.","date":"2025","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/40319028","citation_count":1,"is_preprint":false},{"pmid":"41254209","id":"PMC_41254209","title":"Integration of RASSF1A and TSPYL5 methylation and AFP protein as plasma biomarker for hepatocellular carcinoma diagnosis.","date":"2025","source":"Journal of cancer research and clinical oncology","url":"https://pubmed.ncbi.nlm.nih.gov/41254209","citation_count":1,"is_preprint":false},{"pmid":"42241080","id":"PMC_42241080","title":"TSPYL5 Promotes Triple-Negative Breast Cancer Metastasis by Antagonizing USP10-Mediated PTEN Stabilization to Unleash a ZEB1-Dependent EMT Program.","date":"2026","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/42241080","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.12.30.24319800","title":"Autoantibodies to Arginine-rich Sequences Mimicking Epstein-Barr Virus in Post-COVID and Myalgic Encephalomyelitis/Chronic Fatigue Syndrome","date":"2024-12-31","source":"bioRxiv","url":"https://doi.org/10.1101/2024.12.30.24319800","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13847,"output_tokens":3578,"usd":0.047606,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11132,"output_tokens":4061,"usd":0.078592,"stage2_stop_reason":"end_turn"},"total_usd":0.126198,"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\": 2010,\n      \"finding\": \"TSPYL5 physically interacts with USP7 (ubiquitin-specific protease 7), the deubiquitylase for p53, and reduces USP7 deubiquitylase activity toward p53, resulting in increased p53 ubiquitylation and decreased p53 protein levels. TSPYL5 thereby inhibits p53-target gene activation and overrides p53-dependent proliferation arrest and oncogene-induced senescence.\",\n      \"method\": \"Mass spectrometric analysis of TSPYL5-interacting proteins, co-immunoprecipitation, ubiquitylation assays, cell-based proliferation and senescence assays\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — mass spectrometry identification of binding partner, functional assays (ubiquitylation, proliferation arrest, senescence), multiple orthogonal methods, widely replicated by subsequent studies\",\n      \"pmids\": [\"21170034\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TSPYL5 co-localizes with ALT telomeres in PML bodies and prevents poly-ubiquitination and proteasomal degradation of POT1 (a shelterin component) exclusively in ALT+ cancer cells. USP7 depletion rescued POT1 poly-ubiquitination and loss, indicating that USP7 activates POT1 E3 ubiquitin ligase(s); PML depletion also suppressed POT1 poly-ubiquitination, suggesting interplay between USP7 and PML in triggering POT1 degradation when TSPYL5 is absent.\",\n      \"method\": \"TSPYL5 depletion (siRNA/shRNA), co-localization studies, ubiquitination assays, proteasome inhibitor experiments, USP7 and PML knockdown epistasis experiments\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal epistasis experiments (USP7 KD and PML KD rescuing POT1 loss), multiple orthogonal methods, published in high-impact journal\",\n      \"pmids\": [\"31278054\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"TSPYL5 suppression in A549 lung cancer cells up-regulates PTEN, leading to down-regulation of AKT activation, and also increases p21(WAF1/Cip1) levels, causing cell growth inhibition and sensitization to gamma-radiation. Overexpression of TSPYL5 in H460 cells produced the opposite effects.\",\n      \"method\": \"siRNA knockdown, TSPYL5 overexpression, Western blotting for PTEN/AKT/p21, colony formation assay, gamma-irradiation survival assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — loss-of-function and gain-of-function with defined molecular readouts in two cell lines, single lab\",\n      \"pmids\": [\"20079711\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"TSPYL5 directly binds to the CYP19A1 (aromatase) promoter I.4 and functions as a transcriptional activator of CYP19A1 expression; TSPYL5 knockdown decreased and overexpression increased aromatase expression in MCF-7 cells, lymphoblastoid cell lines, and adipocytes. A putative TSPYL5 binding motif was identified in 43 genes, suggesting broader transcription factor activity.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), TSPYL5 knockdown and overexpression in multiple cell types, reporter assays identifying functional estrogen response element at rs2583506\",\n      \"journal\": \"Molecular endocrinology (Baltimore, Md.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP assay directly demonstrates promoter binding, functional validation across multiple cell types, single lab\",\n      \"pmids\": [\"23518928\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"MUC16 regulates TSPYL5 expression through the JAK2/STAT3/glucocorticoid receptor (GR) axis in lung cancer cells; MUC16 knockdown decreased TSPYL5, and inhibition of STAT3 (Y705) led to decreased GR and TSPYL5, while MUC16-Cter overexpression rescued these signaling proteins and TSPYL5 expression.\",\n      \"method\": \"Stable shRNA knockdown of MUC16, transcriptome analysis, MUC16-Cter overexpression rescue, STAT3 inhibitor treatment, Western blotting\",\n      \"journal\": \"Clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — epistasis established via rescue and inhibitor experiments, transcriptomic identification, single lab\",\n      \"pmids\": [\"28196872\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TSPYL5 promotes human endothelial cell (EC) proliferation, migration, and tube formation by downregulating p53 expression; TSPYL5 depletion-mediated loss of EC functions was rescued by p53 inhibition, and TSPYL5 overexpression blocked adriamycin-induced senescence. In vivo, TSPYL5 overexpression promoted angiogenesis and wound healing.\",\n      \"method\": \"TSPYL5 overexpression and knockdown in HUVECs and hPSC-derived ECs, p53 inhibitor rescue, Matrigel plug angiogenesis assay, mouse wound healing model\",\n      \"journal\": \"Angiogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (p53 inhibitor rescue of TSPYL5-KD phenotype), in vivo validation, single lab\",\n      \"pmids\": [\"30471052\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"AKT phosphorylates TSPYL5 at threonine-120, stabilizing it by inhibiting its ubiquitination. Phosphorylated TSPYL5-pT120 translocates to the nucleus and acts as a transcriptional activator of CSC-associated genes ALDH1 and CD44, while also suppressing PTEN transcription, creating a positive feedback loop (AKT/TSPYL5/PTEN) that maintains cancer stem cell characteristics.\",\n      \"method\": \"Phosphorylation site mutagenesis (T120 substitution), ubiquitination assays, nuclear fractionation, transcriptional reporter assays, ChIP, TSPYL5 knockdown and overexpression in NSCLC cells\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — mutagenesis of phosphorylation site combined with ubiquitination assays, nuclear fractionation, transcriptional assays, and in vivo validation; multiple orthogonal methods\",\n      \"pmids\": [\"34163000\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TSPYL5 localizes to both the cytoplasm and nucleoplasm (including the nucleolus); its nuclear/nucleolar localization is mediated by NLS/NoLS sequences in the N-terminal intrinsically disordered region (residues 4–27), while cytoplasmic localization is regulated by the ordered NAP-like domain (residues 198–382). The N-terminal disordered region (1–198 aa) contributes to exchange dynamics with the nucleoplasm and potential phase separation.\",\n      \"method\": \"Live-cell imaging of TSPYL5-EGFP fusion proteins, FRAP, deletion/truncation constructs, bioinformatics analysis of interactome\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct live imaging and FRAP with functional domain constructs, single lab, no full reconstitution\",\n      \"pmids\": [\"38203210\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Mouse Tspyl5 promotes spermatogonia proliferation by facilitating ubiquitination-mediated degradation of TRP53 (p53), which upregulates Pcna expression and enhances PCNA-mediated DNA replication. Tspyl5 knockout in aged mice caused reduced spermatogonia numbers and spermatozoa, and transcriptomics revealed downregulation of the Pcna-mediated DNA replication pathway.\",\n      \"method\": \"Tspyl5 knockout mouse model, transcriptomic analysis of spermatogonia, ubiquitination assays for TRP53 degradation, Western blotting for PCNA\",\n      \"journal\": \"Reproduction, fertility, and development\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo knockout with defined molecular readout (p53 ubiquitination and PCNA), single lab, mouse model\",\n      \"pmids\": [\"38185096\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TSPYL5 binds G3BP1 and enhances G3BP1 Ser149 phosphorylation, driving G3BP1 nuclear membrane translocation. This recruits p53 for nucleoporin RanBP2, forming a RanBP2-G3BP1-p53 complex that accelerates p53 sumoylation and nuclear export, sequestering p53 in the cytoplasm and suppressing its transcription factor function in neuroblastoma cells.\",\n      \"method\": \"Co-immunoprecipitation, phosphorylation assays, sumoylation assays, nuclear fractionation, complex reconstitution experiments, TSPYL5 overexpression and depletion in neuroblastoma cells\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP and mechanistic epistasis with multiple binding partners identified, single lab, single study\",\n      \"pmids\": [\"40319028\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TSPYL5 sequesters the deubiquitinase USP10, preventing USP10 from stabilizing PTEN, and thereby triggering proteasomal degradation of PTEN in triple-negative breast cancer (TNBC) cells. This TSPYL5-USP10-PTEN axis hyperactivates PI3K/AKT signaling and unleashes a ZEB1-driven EMT/metastatic program, even in cells retaining wild-type PTEN.\",\n      \"method\": \"Co-immunoprecipitation (TSPYL5-USP10 interaction), ubiquitination assays, PTEN stability assays, TSPYL5 overexpression driving orthotopic tumor polymetastasis in animals, single-cell and spatial transcriptomic analyses\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP identifying novel binding partner USP10, functional ubiquitination/stability assays, in vivo metastasis model; single lab, single study\",\n      \"pmids\": [\"42241080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TSPYL5 overexpression in colorectal cancer cells activates endoplasmic reticulum stress (ERS), suppresses cell proliferation, migration, and invasion, and induces apoptosis; upregulation of caspase-1, caspase-3, Bax, ATF4, and CHOP was observed after TSPYL5 overexpression.\",\n      \"method\": \"Transfection of pcDNA3.1-TSPYL5 in HCT116 and HT29 cells, EdU proliferation assay, flow cytometry, Transwell assay, transmission electron microscopy for ERS, Western blotting\",\n      \"journal\": \"Oncology reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — overexpression with cellular phenotype readout and ERS markers, but no defined upstream molecular mechanism; single lab, no pathway placement beyond ERS induction\",\n      \"pmids\": [\"32627024\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TSPYL5 is an intrinsically disordered nucleosome assembly protein (NAP) family member that suppresses p53 function through multiple mechanisms: it inhibits USP7 deubiquitylase activity toward p53 (increasing p53 ubiquitylation and degradation), drives p53 nuclear export via a G3BP1-RanBP2 sumoylation axis, and promotes p53 ubiquitination in spermatogonia to upregulate PCNA-mediated DNA replication; additionally, TSPYL5 protects POT1 from proteasomal degradation in ALT+ cancer cells by antagonizing USP7-dependent ubiquitination, is phosphorylated by AKT at T120 to translocate to the nucleus where it transcriptionally activates CSC genes and suppresses PTEN, sequesters USP10 to trigger PTEN proteasomal degradation in TNBC, and functions as a transcriptional activator of CYP19A1 (aromatase) by directly binding its promoter.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TSPYL5 is a nucleosome assembly protein (NAP)-family member that acts predominantly as a negative regulator of p53, governing proliferation, senescence, and stemness in normal and cancer cells [#0, #5]. Its founding biochemical activity is direct binding to the deubiquitylase USP7, which it inhibits to drive p53 ubiquitylation and degradation, thereby overriding p53-dependent proliferation arrest and oncogene-induced senescence [#0]. TSPYL5 deploys this anti-p53 logic in multiple contexts: in endothelial cells it lowers p53 to promote proliferation, migration, and angiogenesis [#5]; in spermatogonia it facilitates ubiquitination-mediated TRP53 degradation to upregulate PCNA-driven DNA replication [#8]; and in neuroblastoma it binds G3BP1 and enhances its Ser149 phosphorylation, nucleating a RanBP2–G3BP1–p53 complex that accelerates p53 sumoylation and nuclear export to sequester p53 in the cytoplasm [#9]. Beyond p53, TSPYL5 sustains PI3K/AKT signaling by suppressing PTEN — directly downregulating PTEN to lower AKT activity when depleted [#2], and in triple-negative breast cancer sequestering the deubiquitinase USP10 to trigger PTEN proteasomal degradation and a ZEB1-driven EMT program [#10]. AKT in turn phosphorylates TSPYL5 at Thr120, stabilizing it and driving its nuclear translocation where it transcriptionally activates the stem-cell genes ALDH1 and CD44 while repressing PTEN, forming a self-reinforcing AKT/TSPYL5/PTEN feedback loop [#6]. TSPYL5 also functions as a sequence-specific transcriptional activator, binding the CYP19A1 (aromatase) promoter I.4 to upregulate aromatase expression [#3]. In ALT+ cancer cells it localizes to telomeric PML bodies and protects the shelterin component POT1 from USP7- and PML-dependent ubiquitination and degradation [#1]. TSPYL5 is an intrinsically disordered protein whose N-terminal disordered region (residues 4–27) carries NLS/NoLS signals directing nuclear/nucleolar localization, while its ordered NAP-like domain directs cytoplasmic localization [#7].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established the founding molecular mechanism: how TSPYL5 controls cell fate by linking it to the p53 ubiquitylation machinery rather than acting through an unknown route.\",\n      \"evidence\": \"Mass spectrometry of TSPYL5 interactors, co-IP, ubiquitylation assays, and proliferation/senescence assays identifying USP7 inhibition and p53 destabilization\",\n      \"pmids\": [\"21170034\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not define the structural basis of USP7 inhibition\", \"Does not establish whether TSPYL5 acts catalytically or stoichiometrically on USP7\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Connected TSPYL5 to a second tumor-suppressor axis by showing it regulates the PTEN/AKT pathway and the radiation response, broadening its role beyond p53.\",\n      \"evidence\": \"siRNA knockdown and overexpression in lung cancer lines with PTEN/AKT/p21 Western blots, colony formation, and gamma-irradiation survival assays\",\n      \"pmids\": [\"20079711\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which TSPYL5 regulates PTEN not defined at this stage\", \"Single lab, two cell lines\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Demonstrated TSPYL5 has direct sequence-specific transcriptional activity, establishing it as a DNA-binding transactivator rather than solely a protein-interaction scaffold.\",\n      \"evidence\": \"ChIP showing binding to CYP19A1 promoter I.4, knockdown/overexpression across multiple cell types, and reporter assays\",\n      \"pmids\": [\"23518928\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural definition of the DNA-binding interface\", \"The broader 43-gene putative target set remains uncharacterized functionally\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Placed TSPYL5 downstream of an upstream signaling cascade, identifying how its expression is controlled in cancer.\",\n      \"evidence\": \"MUC16 shRNA knockdown, transcriptome analysis, MUC16-Cter rescue, and STAT3 inhibition in lung cancer cells\",\n      \"pmids\": [\"28196872\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect transcriptional control of TSPYL5 not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Extended the anti-p53 function to a physiological vascular context, showing TSPYL5 promotes angiogenesis by suppressing p53.\",\n      \"evidence\": \"Overexpression/knockdown in HUVECs and hPSC-ECs with p53-inhibitor rescue, Matrigel plug, and mouse wound-healing models\",\n      \"pmids\": [\"30471052\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not establish whether the USP7 mechanism operates in endothelial cells\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Revealed a p53-independent role at telomeres, showing TSPYL5 protects POT1 from degradation specifically in ALT+ cancer cells.\",\n      \"evidence\": \"TSPYL5 depletion, PML-body co-localization, ubiquitination and proteasome-inhibitor assays, and USP7/PML knockdown epistasis\",\n      \"pmids\": [\"31278054\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the POT1 E3 ligase activated by USP7 not established\", \"Why protection is ALT-specific not mechanistically resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified the post-translational switch governing TSPYL5 nuclear function, defining a feedback loop that sustains cancer stemness.\",\n      \"evidence\": \"T120 phosphosite mutagenesis, ubiquitination assays, nuclear fractionation, transcriptional reporters and ChIP in NSCLC cells\",\n      \"pmids\": [\"34163000\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct vs indirect PTEN promoter repression not fully separated from ALDH1/CD44 activation\", \"Identity of the E3 ligase counteracted by T120 phosphorylation unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Mapped the determinants of TSPYL5 subcellular partitioning, linking its disordered N-terminus to nuclear/nucleolar targeting and its NAP domain to cytoplasmic retention.\",\n      \"evidence\": \"Live-cell imaging of EGFP fusions, FRAP, and deletion/truncation constructs\",\n      \"pmids\": [\"38203210\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Phase separation only inferred, not reconstituted\", \"Functional consequence of nucleolar pool not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided in vivo genetic evidence for a physiological role, showing Tspyl5 drives spermatogonia proliferation via p53 degradation and PCNA upregulation.\",\n      \"evidence\": \"Tspyl5 knockout mice, spermatogonia transcriptomics, TRP53 ubiquitination assays, and PCNA Westerns\",\n      \"pmids\": [\"38185096\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not confirm the USP7 mechanism operates in spermatogonia\", \"Single lab, mouse model\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a new p53-suppressing mechanism via spatial regulation, showing TSPYL5 drives p53 nuclear export and sumoylation through a G3BP1-RanBP2 axis.\",\n      \"evidence\": \"Co-IP, phosphorylation and sumoylation assays, nuclear fractionation, and complex reconstitution in neuroblastoma cells\",\n      \"pmids\": [\"40319028\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct kinase mediating G3BP1 Ser149 phosphorylation not identified\", \"Single lab, single study\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified USP10 as a new TSPYL5 partner explaining PTEN destabilization in TNBC even with wild-type PTEN, linking TSPYL5 to EMT and metastasis.\",\n      \"evidence\": \"Co-IP of TSPYL5-USP10, ubiquitination/stability assays, orthotopic metastasis models, and single-cell/spatial transcriptomics\",\n      \"pmids\": [\"42241080\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reciprocal validation of the sequestration mechanism limited\", \"Single lab, single study\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how TSPYL5's distinct activities — USP7/USP10 deubiquitylase modulation, sequence-specific transcription, telomeric POT1 protection, and p53 spatial control — are coordinated as a single biochemical mechanism versus context-dependent moonlighting functions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structure of TSPYL5 or its complexes\", \"No unifying biochemical model linking its deubiquitylase-regulatory and transcriptional activities\", \"Whether NAP-family nucleosome assembly activity is biochemically active is untested in the corpus\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [3, 6]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 10]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [10]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [6, 7]},\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 6, 10]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [3, 6]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 10]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 5]}\n    ],\n    \"complexes\": [\"RanBP2-G3BP1-p53 complex\"],\n    \"partners\": [\"USP7\", \"USP10\", \"G3BP1\", \"POT1\", \"RanBP2\", \"p53\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}