{"gene":"S100A16","run_date":"2026-06-10T07:46:28","timeline":{"discoveries":[{"year":2006,"finding":"Human and mouse recombinant S100A16 form homodimers that bind two Ca²⁺ ions exclusively via the C-terminal EF-hand of each subunit (the N-terminal EF-hand is non-functional due to absence of the conserved glutamate); Ca²⁺ binding induces conformational changes detected by Trp fluorescence, and only human S100A16 forms a hydrophobic patch upon Ca²⁺ binding (implicated in target protein recognition). In glioblastoma cells, S100A16 accumulates in nucleoli and translocates to the cytoplasm upon Ca²⁺ stimulation.","method":"Flow dialysis (Ca²⁺ binding), Trp fluorescence spectroscopy, in situ hybridization, immunohistochemistry, immunofluorescence in live cells","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — purified recombinant protein, direct biochemical Ca²⁺ binding assay (flow dialysis), conformational change assay, complemented by cellular localization experiment; multiple orthogonal methods in a single focused study","pmids":["17030513"],"is_preprint":false},{"year":2010,"finding":"NMR and crystal structures of human S100A16 homodimer in apo and Ca²⁺-bound forms show that Ca²⁺ binding produces only minor conformational rearrangement (unlike most S100 proteins), attributed to the absence of the glutamate Ca²⁺ ligand in the N-domain EF-hand and to unusually strong hydrophobic interactions between helices 3 and 4 that stabilize the 'closed' form even after Ca²⁺ binding.","method":"X-ray crystallography and NMR solution structure determination","journal":"Journal of biological inorganic chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — dual structural determination (solid state + solution NMR) in apo and Ca²⁺-bound states with functional interpretation; multiple orthogonal structural methods","pmids":["21046186"],"is_preprint":false},{"year":2011,"finding":"S100A16 promotes adipogenesis in 3T3-L1 preadipocytes: overexpression markedly enhances adipocyte differentiation and proliferation, while RNAi knockdown inhibits both. S100A16 physically interacts with tumor suppressor p53 (co-immunoprecipitation), and S100A16 overexpression inhibits p53-responsive gene expression while knockdown activates them. Elevated intracellular Ca²⁺ causes nuclear exclusion of S100A16. Overexpression also reduces insulin-stimulated glucose uptake and AKT phosphorylation.","method":"3T3-L1 differentiation assay, RNAi knockdown, overexpression, co-immunoprecipitation, Western blot, calcium ionophore treatment","journal":"Endocrinology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal functional experiments (OE and KD), direct Co-IP for p53 interaction, quantitative adipogenesis and signaling assays in a focused mechanistic study","pmids":["21266506"],"is_preprint":false},{"year":2013,"finding":"S100A16 inhibits osteogenesis and stimulates adipogenesis in bone marrow-derived mesenchymal stem cells: S100A16 overexpression increases PPARγ promoter luciferase activity and decreases RUNX2 promoter activity, while knockdown has opposite effects. ERK1/2 phosphorylation is involved in osteogenesis regulation whereas JNK phosphorylation is involved in adipogenesis downstream of S100A16.","method":"BM-MSC differentiation assay, Oil Red O and Alizarin Red S staining, promoter luciferase assay, Western blot for ERK1/2 and JNK phosphorylation, transgenic/knockout mouse-derived cells","journal":"Molecular biology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — promoter luciferase (direct transcriptional readout), functional differentiation assays, signaling assays; single lab with multiple orthogonal readouts","pmids":["23526364"],"is_preprint":false},{"year":2013,"finding":"S100A14 physically interacts with S100A16 (identified by yeast two-hybrid, confirmed by co-immunoprecipitation and co-immunofluorescence). S100A14 overexpression upregulates S100A16 protein without increasing its mRNA, indicating post-transcriptional regulation, whereas S100A16 overexpression does not reciprocally upregulate S100A14 (unidirectional regulation). S100A14/S100A16 degradation is independent of classical proteasomal and lysosomal pathways.","method":"Yeast two-hybrid screen, co-immunoprecipitation, double immunofluorescence, cycloheximide chase, retroviral overexpression and knockdown, RT-PCR","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — interaction identified by Y2H and confirmed by Co-IP and co-IF; post-transcriptional regulation shown by mRNA/protein discordance; replicated across multiple cell lines","pmids":["24086685"],"is_preprint":false},{"year":2014,"finding":"S100A16 overexpression in MCF-7 breast cancer cells upregulates Notch1, ZEB1, and ZEB2 transcription factors, leading to repression of E-cadherin and β-catenin and induction of N-cadherin and vimentin (EMT). Notch1-specific siRNA knockdown reverses EMT induced by S100A16 overexpression, placing Notch1 downstream of S100A16 in this pathway.","method":"Overexpression in MCF-7 cells, siRNA knockdown of Notch1, Western blot, migration/invasion assays, colony formation","journal":"Journal of biomedical science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via siRNA rescue, Western blot pathway analysis; single lab with two complementary experimental approaches","pmids":["25287362"],"is_preprint":false},{"year":2016,"finding":"S100A16 overexpression in DU-145 prostate cancer cells activates AKT and ERK signaling and downregulates p21 and p27. Pharmacological inhibition of AKT (LY294002) or ERK (PD98059) suppresses S100A16-induced clone formation and invasion, placing AKT and ERK downstream of S100A16.","method":"Stable overexpression and shRNA knockdown, Western blot, transwell/wound healing assays, pharmacological inhibitors","journal":"Tumour biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pathway position confirmed by pharmacological epistasis with two inhibitors; single lab, multiple assays","pmids":["27240591"],"is_preprint":false},{"year":2018,"finding":"Brain microvascular endothelial cell (HBMEC) exosomes transfer S100A16 to SCLC cells, causing S100A16 translocation from cytoplasm to nucleus. Elevated S100A16 prevents loss of mitochondrial membrane potential (Δψm) and enhances resistance to apoptosis. This protective effect depends on prohibitin-1 (PHB-1) in the mitochondrial inner membrane, as PHB-1 siRNA delivery into S100A16-overexpressing cells weakens the protective effect.","method":"Exosome isolation by ultracentrifugation, GW4869 inhibitor treatment, Western blot, immunofluorescence, Annexin V/PI apoptosis assay, JC-1 mitochondrial membrane potential assay, PHB-1 siRNA knockdown","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional epistasis via PHB-1 siRNA, multiple readouts (apoptosis, Δψm), exosome transfer mechanism; single lab","pmids":["30183374"],"is_preprint":false},{"year":2019,"finding":"S100A16 interacts with calmodulin (CaM) and regulates hepatic lipid metabolism via the CaM/CAMKK2/AMPK signaling pathway. S100A16 transgenic mice show more severe fatty liver on HFD, while knockdown mice show attenuated steatosis. Overexpression of S100A16 inhibits degradation of 11β-HSD1, a downstream effector of S100A16-induced adipogenesis.","method":"Transgenic and knockout mouse models, HFD feeding, co-immunoprecipitation (S100A16-CaM interaction), Western blot, RNA sequencing, Oil Red O staining","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CaM interaction by Co-IP, in vivo transgenic/KO validation of pathway; single lab with multiple methods","pmids":["31069793"],"is_preprint":false},{"year":2020,"finding":"S100A16 interacts with myosin-9 (identified by mass spectrometry and confirmed functionally) in response to increased Ca²⁺ and TGF-β stimulation, promoting cytoskeleton (F-actin) reorganization and EMT progression in renal tubular epithelial cells. S100A16 overexpression in HK-2 cells increases N-cadherin and vimentin and decreases E-cadherin, and is associated with renal tubulointerstitial fibrosis in both UUO mouse model and patient biopsies.","method":"Mass spectrometry (binding partner identification), immunohistochemistry, UUO mouse model with S100A16 transgenic/heterozygous knockout mice, overexpression and knockdown in HK-2 cells, Western blot, F-actin staining","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-identified interaction with functional validation in cells and in vivo; single lab with multiple orthogonal methods","pmids":["32094322"],"is_preprint":false},{"year":2020,"finding":"S100A16 induces EMT in PDAC cells and promotes metastasis via upregulation of TWIST1 and activation of the STAT3 signaling pathway, as shown by in vitro and in vivo experiments.","method":"siRNA knockdown, overexpression, Western blot, in vivo xenograft, TCGA correlation analysis","journal":"Biochemical pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo loss/gain-of-function with pathway marker analysis; single lab","pmids":["33359364"],"is_preprint":false},{"year":2021,"finding":"S100A16 physically interacts with GRP78 in the endoplasmic reticulum of HK-2 cells (co-immunoprecipitation, immunofluorescence colocalization). S100A16 overexpression causes GRP78 to relocalize from ER to cytoplasm and competitively displaces IRE1α from GRP78, leading to IRE1α phosphorylation and XBP1 splicing (ER stress activation). Calcium chelation with BAPTA-AM blocks both the cytoplasmic colocalization and the upregulation of ER stress markers.","method":"Co-immunoprecipitation, immunofluorescence, lentiviral overexpression, BAPTA-AM chelation, Western blot for ER stress markers","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, immunofluorescence colocalization, competitive binding demonstrated, Ca²⁺-dependence validated pharmacologically; multiple orthogonal methods in single focused study","pmids":["34645789"],"is_preprint":false},{"year":2021,"finding":"S100A16 promotes pancreatic cancer cell proliferation, migration, and invasion via FGF19-dependent activation of AKT and ERK1/2 signaling. S100A16 knockdown induces G2/M cell cycle arrest and apoptosis.","method":"Knockdown and overexpression, in vitro proliferation/migration/invasion assays, in vivo metastasis model, Western blot","journal":"Cell biology and toxicology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — loss and gain of function with FGF19 dependency established; single lab with in vitro and in vivo readouts","pmids":["33389337"],"is_preprint":false},{"year":2021,"finding":"S100A16 promotes gastric cancer cell invasion and EMT via ubiquitination and degradation of ZO-2 (Zonula Occludens-2), a tight junction regulator, identified by proteomic analysis and validated by functional assays.","method":"Proteomics/mass spectrometry (interactome), overexpression and knockdown, functional invasion/migration assays, Western blot for ZO-2 ubiquitination","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-identified interaction, ubiquitination-mediated degradation mechanism shown; single lab","pmids":["34650982"],"is_preprint":false},{"year":2021,"finding":"ADAMTS19 binds cytoplasmic p65 and decreases nuclear phospho-p65 (NF-κB), thereby suppressing S100A16 transcription. S100A16 acts downstream of ADAMTS19 to promote gastric cancer cell migration and invasion, and rescue of S100A16 reverses the suppression caused by ADAMTS19 overexpression.","method":"Co-immunoprecipitation, immunofluorescence, dual-luciferase reporter assay, gain/loss-of-function, transwell assay","journal":"Biomolecules","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and luciferase confirm NF-κB→S100A16 transcriptional axis; epistasis rescue experiment; single lab","pmids":["33921267"],"is_preprint":false},{"year":2022,"finding":"S100A16 promotes Wnt/β-catenin signaling activation in renal interstitial fibroblasts during acute kidney injury by facilitating HRD1 (E3 ubiquitin ligase)-mediated ubiquitination and degradation of GSK3β and CK1α (negative regulators of β-catenin). S100A16 knockout in mice subjected to ischemia-reperfusion injury impedes Wnt/β-catenin activation and restores HGF expression.","method":"S100A16 knockout and overexpression in mice and NRK-49F cells, IRI model, Western blot, ubiquitination assay, ICG-001 Wnt inhibitor","journal":"Cellular and molecular life sciences","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo KO and in vitro OE with convergent mechanistic findings; ubiquitination mechanism identified; pathway position confirmed by pharmacological Wnt inhibitor; multiple orthogonal methods","pmids":["35279748"],"is_preprint":false},{"year":2022,"finding":"S100A16 binds to p53 protein, inducing its degradation; this augments CXCR4 expression, which activates ERK1/2 and AKT signaling, promoting hepatic stellate cell activation and liver fibrosis. S100a16 transgenic mice develop spontaneous liver fibrosis while S100a16 knockout mice are protected.","method":"HSC isolation, S100a16 knockout and transgenic mice, multiple fibrosis models, transcriptome sequencing (RNA-seq), co-immunoprecipitation (S100A16-p53), Western blot","journal":"Metabolism: clinical and experimental","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo transgenic and KO phenotypes, Co-IP for p53 interaction, RNA-seq for pathway identification, multiple disease models; multiple orthogonal methods","pmids":["35914619"],"is_preprint":false},{"year":2024,"finding":"TFAP2B acts as a transcription factor for S100A16 (confirmed by ChIP and luciferase reporter assay). HIF-1α transcriptionally regulates HRD1 (confirmed by ChIP and luciferase reporter assay) within the S100A16→HRD1→GSK3β/CK1α pathway in renal hypoxia injury. S100A16 deletion attenuates HIF-1α upregulation during IRI.","method":"ChIP assay, luciferase reporter assay, S100A16 knockout rat cell line (NRK-52E), hypoxia/reoxygenation model, Western blot","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — ChIP and luciferase reporter for transcription factor binding; single lab, two orthogonal methods for each transcriptional relationship","pmids":["38710691"],"is_preprint":false},{"year":2024,"finding":"During myocardial ischemia/reperfusion injury, VDAC1 upregulation activates NF-κB/p65, which binds to the S100A16 promoter to drive S100A16 transcription. S100A16 then interacts with calmodulin (CaM) in response to elevated Ca²⁺ to activate the CAMKK2/AMPK pathway, contributing to inflammation and ROS production. Adenovirus-mediated S100A16 inhibition improves cardiac function and reduces infarct size.","method":"In vivo cardiac I/R model (ligation/release), H/R cell model, adenovirus-mediated S100A16 inhibition, ChIP-like and luciferase assays for NF-κB promoter binding, Western blot, co-immunoprecipitation (S100A16-CaM)","journal":"European journal of pharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for CaM interaction, promoter binding confirmed, in vivo and in vitro concordant results; single lab","pmids":["39613175"],"is_preprint":false},{"year":2024,"finding":"S100A16 binds to MOV10 RNA helicase (co-immunoprecipitation) and positively modulates MOV10 expression in lung adenocarcinoma cells. MOV10 in turn stabilizes ITGA3 mRNA (RNA immunoprecipitation, actinomycin D mRNA stability assay), thereby regulating ECM-receptor interaction signaling and promoting malignant properties. MOV10 overexpression partially reverses the suppressive effects of S100A16 knockdown.","method":"Co-immunoprecipitation (S100A16-MOV10), RNA immunoprecipitation (MOV10-ITGA3 mRNA), actinomycin D mRNA stability assay, knockdown and overexpression, Western blot","journal":"Molecular medicine reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and RIP with mRNA stability assay establish mechanism; epistasis rescue; single lab with three orthogonal methods","pmids":["39450567"],"is_preprint":false},{"year":2025,"finding":"S100A16 knockdown in HeLa and SiHa cervical cancer cells inhibits cell migration. S100A16 regulates RPN2 (ribophorin II) expression via phospho-STAT3, which in turn modulates phospho-GSK3β to activate β-catenin/TCF signaling. S100A16 silencing reduces nuclear translocation of β-catenin (shown by nuclear/cytosolic fractionation). RPN2 overexpression reverses the migration suppression caused by S100A16 knockdown.","method":"siRNA knockdown, RNA sequencing, nuclear/cytosolic fractionation, Western blot, overexpression rescue experiment, wound-healing migration assay","journal":"Biochimica et biophysica acta. Molecular cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RNA-seq for downstream target, fractionation for β-catenin localization, rescue epistasis; single lab with multiple orthogonal methods","pmids":["40907797"],"is_preprint":false},{"year":2025,"finding":"S100A16 is enriched in nucleoli of metastatic breast cancer cells (nucleolar proteomics). ChIP-MS shows S100A16 associates at rDNA loci with RPA194 (catalytic subunit of RNA Polymerase I). Loss of S100A16 disrupts RNA Polymerase I activation and rRNA synthesis, reverses EMT, inhibits invasion, and reduces metastatic incidence in animal models.","method":"Nucleolar proteomics, ChIP-MS (rDNA loci, RPA194), loss-of-function in vitro and in vivo metastasis assays, rRNA synthesis assay","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 1 / Strong — ChIP-MS directly links S100A16 to rDNA/RNA Pol I; nucleolar proteomics for localization; in vitro and in vivo functional validation; multiple orthogonal methods in single study","pmids":["40846689"],"is_preprint":false},{"year":2026,"finding":"S100A14 physically interacts with S100A16 (confirmed by Co-IP), stabilizes S100A16 protein via post-translational modification without transcriptional regulation (CHX chase and mRNA analysis). The S100A14/S100A16 complex reduces p53 protein stability and inhibits p53 transcriptional activity and downstream p21 expression. Knockdown of S100A14 restores p53 function, consistent with the previously described S100A16-p53 interaction.","method":"Co-immunoprecipitation, CHX chase assay, dual-luciferase assay (p53 transcriptional activity), siRNA knockdown, Western blot","journal":"Oncology research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, protein stability assay, and transcriptional activity assay confirm mechanism; single lab, multiple orthogonal methods","pmids":["41799516"],"is_preprint":false},{"year":2026,"finding":"CRYAB lysine 92 lactylation (CRYAB_K92la) enhances S100A16 transcription, stabilizes S100A16 protein by inhibiting its degradation, and strengthens CRYAB-S100A16 binding. Upregulated S100A16 activates RAGE signaling, which promotes lactate production, forming a glycolysis-fibrosis positive feedback loop in ligamentum flavum hypertrophy.","method":"Mass spectrometry (lactylation identification), CRYAB_K92R lactylation-blocking mutation, co-immunoprecipitation (CRYAB-S100A16), promoter assay (S100A16 transcription), Western blot, in vitro fibrosis assays","journal":"Communications biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — PTM identified by MS, functional mutation (K92R), Co-IP for protein-protein interaction; single lab with multiple methods","pmids":["42098255"],"is_preprint":false},{"year":2026,"finding":"S100A16 preferentially interacts with saturated phospholipids with short acyl chains and zwitterionic polar head groups, and with cholesterol-enriched membrane domains (lipid raft-like), as determined using Langmuir monolayer biophysical assays. These interactions are calcium-sensitive and suggest S100A16 associates with plasma membrane and nascent disk membranes in photoreceptor outer segments.","method":"Langmuir monolayer model, surface tensiometry","journal":"Archives of biochemistry and biophysics","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro reconstitution biophysical assay with defined lipid compositions; single lab, single method","pmids":["42203137"],"is_preprint":false},{"year":2022,"finding":"S100A16 knockdown in renal cancer cells inhibits proliferation and migration and reduces VEGF, VEGFR2, and phospho-AKT expression, implicating S100A16 in the VEGF/VEGFR2/PI3K-AKT signaling axis in renal cell carcinoma.","method":"siRNA knockdown, Western blot, CCK8, wound healing/transwell assays","journal":"Contrast media & molecular imaging","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, Western blot for pathway markers after knockdown; no direct binding or reconstitution experiments","pmids":["36176934"],"is_preprint":false},{"year":2022,"finding":"S100A16 knockdown in bladder cancer cells suppresses the AKT/Bcl-2 pathway to promote apoptosis and sensitizes cells to mitomycin C. The EMT-associated transcription factor Snail regulates S100A16 expression in resistant bladder cancer cells.","method":"Proteomics (LC-MS/MS) for initial identification, siRNA knockdown, Western blot, CCK8 viability assay","journal":"Cancer management and research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — Snail-S100A16 regulatory link inferred from expression correlation after proteomics; siRNA knockdown with pathway Western blot; no direct binding assay; single lab","pmids":["31118765"],"is_preprint":false},{"year":2023,"finding":"S100A16 deletion protects mice against alcoholic liver lipid accumulation and inflammation by upregulating mesencephalic astrocyte-derived neurotrophic factor (MANF), which inhibits ER stress signaling. MANF silencing suppressed the inhibitory effect of S100a16 knockout on ethanol-induced lipid droplet accumulation in primary hepatocytes.","method":"S100a16 knockout and transgenic mice, Gao-binge alcohol feeding model, MANF siRNA, Western blot, lipid staining, ELISA","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo KO and transgenic with epistasis rescue (MANF siRNA); concordant in vitro and in vivo; single lab","pmids":["37928262"],"is_preprint":false},{"year":2022,"finding":"Co-silencing of S100A16 and HSP27 in placenta-derived multipotent stem cells (PDMCs) without chemical induction drives differentiation into functional astrocytes (confirmed by morphology, neural marker immunofluorescence, electrophysiology, and Ca²⁺ influx measurement), indicating S100A16 suppresses astrocyte differentiation.","method":"siRNA knockdown (combinatorial), immunofluorescence quantification, electrophysiology, Ca²⁺ influx assay, transcriptomic/proteomic cross-comparison","journal":"Stem cell reviews and reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional differentiation with electrophysiological confirmation; multiple orthogonal validation methods; single lab","pmids":["35061207"],"is_preprint":false},{"year":2023,"finding":"S100A16 knockdown in nephroblastoma cells inhibits proliferation, invasion, migration, and angiogenesis. S100A16 physically interacts with DEPDC1 (co-immunoprecipitation), and DEPDC1 overexpression partially reverses the suppressive effects of S100A16 interference, including suppression of PI3K/Akt/mTOR pathway activation.","method":"Co-immunoprecipitation, siRNA knockdown, overexpression rescue, CCK8, Transwell, tube formation assay, Western blot","journal":"Polish journal of pathology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP, epistasis by overexpression rescue; single lab, limited mechanistic depth","pmids":["37955537"],"is_preprint":false},{"year":2018,"finding":"In cancer stem-like spheroids of Yumoto cervical carcinoma cells, S100A16 knockdown decreases Oct4 and Nanog mRNA and protein expression and reduces sphere size. S100A16 knockdown also restores p53 protein that was diminished during sphere formation. Proteasome inhibitor lactacystin decreases Oct4 and Nanog protein but does not affect S100A16 protein, indicating S100A16 maintains Oct4/Nanog via a proteasome-dependent mechanism without being itself a proteasomal target.","method":"Sphere formation assay, siRNA knockdown, RT-PCR, Western blot, proteasome inhibitor (lactacystin) treatment","journal":"Oncology letters","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, indirect mechanistic inference from proteasome inhibitor; limited pathway resolution","pmids":["29928366"],"is_preprint":false}],"current_model":"S100A16 is a homodimeric EF-hand calcium-binding protein that senses Ca²⁺ through its C-terminal EF-hand (the N-terminal site is non-functional), undergoes Ca²⁺-dependent conformational changes and nucleolar-to-cytoplasmic translocation, and executes diverse cellular functions through multiple experimentally confirmed molecular interactions: it binds and inhibits p53 (promoting adipogenesis and cancer progression), interacts with calmodulin to regulate the CaM/CAMKK2/AMPK pathway (controlling lipid metabolism and myocardial stress responses), associates with GRP78 in the ER to competitively release IRE1α and activate ER stress via IRE1α/XBP1, interacts with myosin-9 to drive cytoskeletal reorganization and EMT, promotes HRD1-mediated ubiquitination and degradation of GSK3β and CK1α to activate Wnt/β-catenin signaling, localizes to nucleoli where it associates with RNA Polymerase I at rDNA loci to drive rRNA synthesis and support metastasis, and is transcriptionally regulated by TFAP2B, NF-κB/p65, and BRD4-dependent H3K27 acetylation."},"narrative":{"mechanistic_narrative":"S100A16 is a homodimeric EF-hand calcium-binding protein that senses Ca²⁺ exclusively through the C-terminal EF-hand of each subunit—the N-terminal site being non-functional owing to loss of the conserved glutamate ligand—and translates this into Ca²⁺-dependent conformational change, subcellular redistribution, and target engagement that collectively drive differentiation, metabolic, and pro-metastatic programs [PMID:17030513, PMID:21046186]. Unlike most S100 proteins, Ca²⁺ binding produces only a minor structural rearrangement because strong hydrophobic helix 3–4 interactions stabilize the closed conformation, yet human S100A16 still forms a Ca²⁺-induced hydrophobic patch for target recognition [PMID:17030513, PMID:21046186], and it accumulates in nucleoli, translocating to the cytoplasm or nucleus upon Ca²⁺ stimulation [PMID:17030513, PMID:21266506]. A recurrent mechanistic theme is inhibition of the tumor suppressor p53: S100A16 binds p53 and represses its transcriptional output, promoting adipogenesis and, through p53 degradation and downstream CXCR4/ERK/AKT signaling, hepatic stellate cell activation and liver fibrosis in transgenic and knockout mouse models [PMID:21266506, PMID:35914619]. S100A16 controls lipid metabolism and stress responses through calmodulin binding and activation of the CaM/CAMKK2/AMPK axis [PMID:31069793, PMID:39613175], and engages the ER chaperone GRP78 to competitively displace IRE1α, triggering IRE1α/XBP1-mediated ER stress in a Ca²⁺-dependent manner [PMID:34645789]. In cancer and fibrosis it drives epithelial–mesenchymal transition and invasion via multiple effectors—myosin-9-dependent cytoskeletal reorganization [PMID:32094322], and ubiquitin-mediated turnover of negative regulators, facilitating HRD1-mediated degradation of GSK3β and CK1α to activate Wnt/β-catenin signaling [PMID:35279748]. In metastatic breast cancer it localizes to nucleoli and associates with RNA Polymerase I (RPA194) at rDNA loci to support rRNA synthesis, EMT, and metastasis [PMID:40846689]. S100A16 is itself stabilized by its paralog S100A14 through a post-translational, non-transcriptional mechanism, with the S100A14/S100A16 complex amplifying p53 suppression [PMID:24086685, PMID:41799516], and its transcription is governed by TFAP2B and NF-κB/p65 [PMID:38710691, PMID:39613175].","teleology":[{"year":2006,"claim":"Established S100A16 as a genuine EF-hand Ca²⁺ sensor and defined the structural basis of its binding, answering whether and how this S100 family member responds to calcium.","evidence":"Flow dialysis Ca²⁺ binding, Trp fluorescence, and live-cell localization of recombinant human/mouse protein in glioblastoma cells","pmids":["17030513"],"confidence":"High","gaps":["Did not identify physiological Ca²⁺-dependent target proteins","Functional consequence of nucleolar-to-cytoplasmic translocation unknown"]},{"year":2010,"claim":"Resolved why S100A16 behaves atypically among S100 proteins by showing Ca²⁺ binding causes only minimal conformational change, refining the structural model.","evidence":"X-ray crystallography and NMR solution structures of apo and Ca²⁺-bound homodimer","pmids":["21046186"],"confidence":"High","gaps":["Structural basis of specific target recognition not resolved","No co-structure with a partner protein"]},{"year":2011,"claim":"Connected S100A16 to adipogenesis and identified p53 as a direct binding partner whose activity it represses, providing the first molecular partner and a tumor-suppressor link.","evidence":"3T3-L1 differentiation with reciprocal overexpression/knockdown, p53 Co-IP, and Ca²⁺-ionophore-induced nuclear exclusion","pmids":["21266506"],"confidence":"High","gaps":["Binding interface and Ca²⁺-dependence of p53 interaction not mapped","Mechanism by which binding represses p53 transcriptional output unclear"]},{"year":2013,"claim":"Showed S100A16 reciprocally controls the adipogenic/osteogenic fate switch in mesenchymal stem cells through ERK1/2 and JNK signaling, extending its role to lineage commitment.","evidence":"BM-MSC differentiation, PPARγ/RUNX2 promoter luciferase, and phospho-MAPK Western blots","pmids":["23526364"],"confidence":"Medium","gaps":["Direct molecular link between S100A16 and MAPK activation not established","No in vivo skeletal phenotype tested"]},{"year":2013,"claim":"Identified the paralog S100A14 as a direct interactor that post-transcriptionally stabilizes S100A16, revealing a regulatory dimerization mechanism.","evidence":"Yeast two-hybrid, Co-IP, co-IF, and cycloheximide chase across multiple cell lines","pmids":["24086685"],"confidence":"High","gaps":["Degradation pathway (non-proteasomal, non-lysosomal) not identified","Functional consequence of the complex not yet defined here"]},{"year":2014,"claim":"Placed S100A16 upstream of EMT transcription factors in breast cancer, defining a pro-invasive transcriptional program.","evidence":"MCF-7 overexpression with Notch1 siRNA rescue and EMT marker analysis","pmids":["25287362"],"confidence":"Medium","gaps":["Mechanism by which S100A16 upregulates Notch1/ZEB unknown","Single cell line"]},{"year":2016,"claim":"Extended S100A16 pro-tumor signaling to prostate cancer via AKT/ERK activation and cell-cycle inhibitor downregulation.","evidence":"DU-145 overexpression/knockdown with LY294002 and PD98059 pharmacological epistasis","pmids":["27240591"],"confidence":"Medium","gaps":["No direct binding partner upstream of AKT/ERK identified","Single cell line"]},{"year":2018,"claim":"Demonstrated intercellular transfer of S100A16 via exosomes confers apoptosis resistance through mitochondrial PHB-1, introducing a non-cell-autonomous mechanism.","evidence":"HBMEC-to-SCLC exosome transfer, JC-1 membrane-potential assay, and PHB-1 siRNA epistasis","pmids":["30183374"],"confidence":"Medium","gaps":["Direct S100A16-PHB-1 binding not shown","Mechanism of nuclear translocation upon transfer unclear"]},{"year":2019,"claim":"Identified calmodulin as a binding partner linking S100A16 to the CaM/CAMKK2/AMPK pathway controlling hepatic lipid metabolism in vivo.","evidence":"S100A16 transgenic/knockout mice on HFD, CaM Co-IP, RNA-seq","pmids":["31069793"],"confidence":"Medium","gaps":["Ca²⁺-dependence of CaM interaction not formally tested here","Direct effect on AMPK vs indirect not separated"]},{"year":2020,"claim":"Identified myosin-9 as a Ca²⁺/TGF-β-induced interactor driving cytoskeletal reorganization and EMT in renal fibrosis, linking S100A16 to fibrotic disease.","evidence":"Mass spectrometry interactome, UUO mouse model with transgenic/knockout mice, HK-2 functional assays","pmids":["32094322"],"confidence":"Medium","gaps":["Binding interface with myosin-9 not mapped","Whether myosin-9 binding is direct not biochemically proven"]},{"year":2020,"claim":"Showed S100A16 drives PDAC metastasis through TWIST1 and STAT3, generalizing its EMT-promoting role to pancreatic cancer.","evidence":"Loss/gain-of-function in vitro and xenograft with TCGA correlation","pmids":["33359364"],"confidence":"Medium","gaps":["No direct molecular partner linking S100A16 to STAT3","Mechanism of TWIST1 induction unknown"]},{"year":2021,"claim":"Defined a Ca²⁺-dependent ER stress mechanism in which S100A16 binds GRP78 and competitively displaces IRE1α to activate IRE1α/XBP1 signaling.","evidence":"Reciprocal Co-IP, IF colocalization, competitive binding, BAPTA-AM Ca²⁺ chelation in HK-2 cells","pmids":["34645789"],"confidence":"High","gaps":["Stoichiometry of GRP78 displacement not quantified","Whether mechanism operates outside renal cells untested"]},{"year":2021,"claim":"Linked S100A16 to FGF19-dependent AKT/ERK signaling driving pancreatic cancer proliferation and survival.","evidence":"Knockdown/overexpression, cell-cycle and apoptosis assays, in vivo metastasis model","pmids":["33389337"],"confidence":"Medium","gaps":["Mechanism connecting S100A16 to FGF19 unclear","No direct binding partner identified"]},{"year":2021,"claim":"Identified ZO-2 as a degradation target, showing S100A16 promotes gastric cancer invasion by ubiquitin-mediated turnover of a tight-junction regulator.","evidence":"Proteomic interactome, functional assays, ZO-2 ubiquitination Western blot","pmids":["34650982"],"confidence":"Medium","gaps":["E3 ligase mediating ZO-2 ubiquitination not identified","Direct vs indirect role of S100A16 in degradation unclear"]},{"year":2021,"claim":"Established an NF-κB-driven transcriptional axis upstream of S100A16 suppressed by ADAMTS19 in gastric cancer.","evidence":"Co-IP, dual-luciferase reporter, and S100A16 rescue epistasis","pmids":["33921267"],"confidence":"Medium","gaps":["Direct p65 binding to S100A16 promoter assumed not fully mapped","Single cancer context"]},{"year":2022,"claim":"Resolved how S100A16 activates Wnt/β-catenin by facilitating HRD1-mediated ubiquitination and degradation of GSK3β and CK1α, providing a defined enzymatic mechanism in kidney injury.","evidence":"S100A16 knockout/overexpression mice and NRK-49F cells, IRI model, ubiquitination assay, ICG-001 Wnt inhibitor","pmids":["35279748"],"confidence":"High","gaps":["Whether S100A16 directly scaffolds HRD1 onto substrates not biochemically shown","Ca²⁺-dependence of this mechanism untested"]},{"year":2022,"claim":"Mechanistically tied S100A16-mediated p53 degradation to CXCR4/ERK/AKT signaling in hepatic stellate cell activation and liver fibrosis with strong in vivo support.","evidence":"Transgenic and knockout mice across fibrosis models, p53 Co-IP, RNA-seq","pmids":["35914619"],"confidence":"High","gaps":["E3 ligase for p53 degradation not identified","Direct CXCR4 regulation mechanism not mapped"]},{"year":2024,"claim":"Identified TFAP2B as a direct transcription factor for S100A16 and integrated HIF-1α/HRD1 regulation into the renal hypoxia pathway.","evidence":"ChIP and luciferase reporter assays, S100A16 knockout NRK-52E cells, H/R model","pmids":["38710691"],"confidence":"Medium","gaps":["Relative contribution of TFAP2B vs NF-κB in different tissues unclear","Single experimental system"]},{"year":2024,"claim":"Showed VDAC1/NF-κB-driven S100A16 transcription and CaM/CAMKK2/AMPK activation contribute to myocardial ischemia/reperfusion injury, defining a cardiac stress axis.","evidence":"Cardiac I/R and H/R models, adenoviral inhibition, promoter binding and CaM Co-IP","pmids":["39613175"],"confidence":"Medium","gaps":["Direct VDAC1-S100A16 relationship not established","Ca²⁺-dependence of CaM interaction in cardiac context not isolated"]},{"year":2024,"claim":"Revealed an RNA-regulatory function in which S100A16 binds the MOV10 helicase to stabilize ITGA3 mRNA and promote lung adenocarcinoma malignancy.","evidence":"Co-IP, RIP, actinomycin D mRNA stability assay, knockdown/overexpression rescue","pmids":["39450567"],"confidence":"Medium","gaps":["Direct vs indirect MOV10 binding not fully resolved","Whether S100A16 itself binds RNA untested"]},{"year":2025,"claim":"Defined a nucleolar function: S100A16 associates with RNA Polymerase I (RPA194) at rDNA loci to drive rRNA synthesis and support breast cancer metastasis, explaining its long-observed nucleolar localization.","evidence":"Nucleolar proteomics, ChIP-MS at rDNA loci, loss-of-function in vitro and in vivo metastasis assays","pmids":["40846689"],"confidence":"High","gaps":["How S100A16 is recruited to rDNA not defined","Ca²⁺-dependence of RNA Pol I association untested"]},{"year":2025,"claim":"Connected S100A16 to RPN2/STAT3/GSK3β-dependent β-catenin nuclear translocation driving cervical cancer migration.","evidence":"siRNA knockdown, RNA-seq, nuclear/cytosolic fractionation, RPN2 rescue","pmids":["40907797"],"confidence":"Medium","gaps":["Direct partner linking S100A16 to RPN2 expression unknown","Single cancer type"]},{"year":2026,"claim":"Confirmed the S100A14/S100A16 complex amplifies p53 suppression through post-translational stabilization of S100A16, unifying paralog interaction with p53 regulation.","evidence":"Co-IP, CHX chase, dual-luciferase p53 activity assay, S100A14 knockdown","pmids":["41799516"],"confidence":"Medium","gaps":["Identity of the stabilizing post-translational modification unknown","Stoichiometry of the complex on p53 not defined"]},{"year":2026,"claim":"Showed CRYAB K92 lactylation enhances S100A16 transcription and stability and strengthens CRYAB-S100A16 binding, embedding S100A16 in a glycolysis-fibrosis RAGE feedback loop.","evidence":"Mass spectrometry, K92R lactylation-blocking mutant, Co-IP, promoter assay, fibrosis assays","pmids":["42098255"],"confidence":"Medium","gaps":["Direct RAGE binding by S100A16 not shown","Single disease context"]},{"year":2026,"claim":"Characterized Ca²⁺-sensitive S100A16 interactions with saturated short-chain phospholipids and cholesterol-rich membrane domains, suggesting a membrane-association function.","evidence":"Langmuir monolayer biophysical assays with defined lipid compositions","pmids":["42203137"],"confidence":"Medium","gaps":["Single in vitro biophysical method","Membrane association not validated in cells"]},{"year":null,"claim":"How a single small Ca²⁺ sensor with minimal Ca²⁺-induced conformational change selects among its many reported partners (p53, CaM, GRP78, myosin-9, MOV10, RNA Pol I) in different cell types, and whether a unifying biochemical rule governs target choice, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No co-structure of S100A16 with any partner","Ca²⁺-dependence not uniformly tested across interactions","Tissue-specific determinants of partner selection unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[13,15]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,11,16]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[9]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[24]}],"localization":[{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[0,21]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,2]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[11]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,7]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[24]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[8,15,18]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[11,27]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[2,3]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[21]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[16,9]}],"complexes":["S100A14/S100A16 complex"],"partners":["S100A14","TP53","CALM1","HSPA5","MYH9","MOV10","CRYAB","DEPDC1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q96FQ6","full_name":"Protein S100-A16","aliases":["Aging-associated gene 13 protein","Protein S100-F","S100 calcium-binding protein A16"],"length_aa":103,"mass_kda":11.8,"function":"Calcium-binding protein. Binds one calcium ion per monomer (PubMed:17030513). Can promote differentiation of adipocytes (in vitro) (By similarity). Overexpression in preadipocytes increases their proliferation, enhances adipogenesis and reduces insulin-stimulated glucose uptake (By similarity)","subcellular_location":"Nucleus, nucleolus; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q96FQ6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/S100A16","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/S100A16","total_profiled":1310},"omim":[{"mim_id":"617437","title":"S100 CALCIUM-BINDING PROTEIN A16; S100A16","url":"https://www.omim.org/entry/617437"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"esophagus","ntpm":2382.1}],"url":"https://www.proteinatlas.org/search/S100A16"},"hgnc":{"alias_symbol":["S100F","DT1P1A7","MGC17528"],"prev_symbol":[]},"alphafold":{"accession":"Q96FQ6","domains":[{"cath_id":"1.10.238.10","chopping":"8-102","consensus_level":"high","plddt":83.6534,"start":8,"end":102}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96FQ6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96FQ6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96FQ6-F1-predicted_aligned_error_v6.png","plddt_mean":81.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=S100A16","jax_strain_url":"https://www.jax.org/strain/search?query=S100A16"},"sequence":{"accession":"Q96FQ6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96FQ6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96FQ6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96FQ6"}},"corpus_meta":[{"pmid":"30183374","id":"PMC_30183374","title":"Brain 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of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27240591","citation_count":44,"is_preprint":false},{"pmid":"33389337","id":"PMC_33389337","title":"S100A16 promotes metastasis and progression of pancreatic cancer through FGF19-mediated AKT and ERK1/2 pathways.","date":"2021","source":"Cell biology and toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/33389337","citation_count":42,"is_preprint":false},{"pmid":"34645789","id":"PMC_34645789","title":"The interaction of S100A16 and GRP78 actives endoplasmic reticulum stress-mediated through the IRE1α/XBP1 pathway in renal tubulointerstitial fibrosis.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/34645789","citation_count":33,"is_preprint":false},{"pmid":"33359364","id":"PMC_33359364","title":"S100A16 induces epithelial-mesenchymal transition in human PDAC cells and is a new therapeutic target for pancreatic cancer treatment that 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and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34650982","citation_count":18,"is_preprint":false},{"pmid":"29928366","id":"PMC_29928366","title":"S100A16 up-regulates Oct4 and Nanog expression in cancer stem-like cells of Yumoto human cervical carcinoma cells.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/29928366","citation_count":18,"is_preprint":false},{"pmid":"24501224","id":"PMC_24501224","title":"Estrogen suppresses adipogenesis by inhibiting S100A16 expression.","date":"2014","source":"Journal of molecular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/24501224","citation_count":17,"is_preprint":false},{"pmid":"37509106","id":"PMC_37509106","title":"An Update on S100A16 in Human Cancer.","date":"2023","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/37509106","citation_count":16,"is_preprint":false},{"pmid":"38710691","id":"PMC_38710691","title":"HIF-1α participates in the regulation of S100A16-HRD1-GSK3β/CK1α pathway in renal hypoxia injury.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/38710691","citation_count":14,"is_preprint":false},{"pmid":"31894756","id":"PMC_31894756","title":"S100A16 Regulates HeLa Cell through the Phosphatidylinositol 3 Kinase (PI3K)/AKT Signaling Pathway.","date":"2020","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/31894756","citation_count":14,"is_preprint":false},{"pmid":"36176934","id":"PMC_36176934","title":"Calbindin S100A16 Promotes Renal Cell Carcinoma Progression and Angiogenesis via the VEGF/VEGFR2 Signaling Pathway.","date":"2022","source":"Contrast media & molecular imaging","url":"https://pubmed.ncbi.nlm.nih.gov/36176934","citation_count":13,"is_preprint":false},{"pmid":"36224471","id":"PMC_36224471","title":"An inhibitor of BRD4, GNE987, inhibits the growth of glioblastoma cells by targeting C-Myc and S100A16.","date":"2022","source":"Cancer chemotherapy and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/36224471","citation_count":13,"is_preprint":false},{"pmid":"31399502","id":"PMC_31399502","title":"S100A16-induced adipogenesis is associated with up-regulation of 11 β-hydroxysteroid dehydrogenase type 1 (11β-HSD1).","date":"2019","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/31399502","citation_count":11,"is_preprint":false},{"pmid":"30916375","id":"PMC_30916375","title":"S100A16 suppresses the growth and survival of leukaemia cells and correlates with relapse and relapse free survival in adults with Philadelphia chromosome-negative B-cell acute lymphoblastic leukaemia.","date":"2019","source":"British journal of haematology","url":"https://pubmed.ncbi.nlm.nih.gov/30916375","citation_count":11,"is_preprint":false},{"pmid":"21871643","id":"PMC_21871643","title":"S100A16 mediation of weight gain attenuation induced by dietary 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ADAMTS19 Suppresses Cell Migration and Invasion by Targeting S100A16 via the NF-κB Pathway in Human Gastric Cancer. Biomolecules 2021, 11, 561.","date":"2025","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/41594721","citation_count":3,"is_preprint":false},{"pmid":"38733818","id":"PMC_38733818","title":"S100A16 is a potential target for reshaping the tumor microenvironment in the hypoxic context of liver cancer.","date":"2024","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/38733818","citation_count":2,"is_preprint":false},{"pmid":"38520747","id":"PMC_38520747","title":"SPDEF drives pancreatic adenocarcinoma progression via transcriptional upregulation of S100A16 and activation of the PI3K/AKT signaling pathway.","date":"2024","source":"Biomolecules & biomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/38520747","citation_count":1,"is_preprint":false},{"pmid":"40907797","id":"PMC_40907797","title":"S100A16 knockdown reduces RPN2 expression and inhibits β-catenin/TCF signaling, leading to suppressed metastasis in cervical cancer cells.","date":"2025","source":"Biochimica et biophysica acta. Molecular cell research","url":"https://pubmed.ncbi.nlm.nih.gov/40907797","citation_count":0,"is_preprint":false},{"pmid":"41722834","id":"PMC_41722834","title":"Integrative single-cell and spatial transcriptomic reveals S100A16+ tumor endothelial cells drive angiogenesis and immunosuppression in hepatocellular carcinoma.","date":"2026","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/41722834","citation_count":0,"is_preprint":false},{"pmid":"40846689","id":"PMC_40846689","title":"Nucleolar proteomics identifies S100A16 as a key nucleolar protein driving breast cancer metastasis.","date":"2025","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/40846689","citation_count":0,"is_preprint":false},{"pmid":"40688271","id":"PMC_40688271","title":"Validation of S100A16 as an asthma biomarker and its role in IL-13-induced bronchial epithelial cell injury.","date":"2025","source":"Journal of thoracic disease","url":"https://pubmed.ncbi.nlm.nih.gov/40688271","citation_count":0,"is_preprint":false},{"pmid":"41799516","id":"PMC_41799516","title":"S100A14 Facilitates Pancreatic Cancer Progression via S100A16-Mediated p53 Suppression.","date":"2026","source":"Oncology research","url":"https://pubmed.ncbi.nlm.nih.gov/41799516","citation_count":0,"is_preprint":false},{"pmid":"41868093","id":"PMC_41868093","title":"Integrated bioinformatics and immunohistochemical analysis reveal that S100A16 is correlated with mutational burden, immune evasion, and P53 expression in gastric adenocarcinoma.","date":"2026","source":"International journal of clinical and experimental pathology","url":"https://pubmed.ncbi.nlm.nih.gov/41868093","citation_count":0,"is_preprint":false},{"pmid":"42203137","id":"PMC_42203137","title":"Effect of calcium and cholesterol on membrane binding and localization of the S100A16 protein.","date":"2026","source":"Archives of biochemistry and biophysics","url":"https://pubmed.ncbi.nlm.nih.gov/42203137","citation_count":0,"is_preprint":false},{"pmid":"42098255","id":"PMC_42098255","title":"CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop.","date":"2026","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/42098255","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":32462,"output_tokens":8500,"usd":0.112443,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":18377,"output_tokens":7153,"usd":0.135355,"stage2_stop_reason":"end_turn"},"total_usd":0.247798,"stage1_batch_id":"msgbatch_013LWZZT6MWaqB9qyTSeQBmU","stage2_batch_id":"msgbatch_0194ZzQaTGNpruoNJBbEMbt3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2006,\n      \"finding\": \"Human and mouse recombinant S100A16 form homodimers that bind two Ca²⁺ ions exclusively via the C-terminal EF-hand of each subunit (the N-terminal EF-hand is non-functional due to absence of the conserved glutamate); Ca²⁺ binding induces conformational changes detected by Trp fluorescence, and only human S100A16 forms a hydrophobic patch upon Ca²⁺ binding (implicated in target protein recognition). In glioblastoma cells, S100A16 accumulates in nucleoli and translocates to the cytoplasm upon Ca²⁺ stimulation.\",\n      \"method\": \"Flow dialysis (Ca²⁺ binding), Trp fluorescence spectroscopy, in situ hybridization, immunohistochemistry, immunofluorescence in live cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — purified recombinant protein, direct biochemical Ca²⁺ binding assay (flow dialysis), conformational change assay, complemented by cellular localization experiment; multiple orthogonal methods in a single focused study\",\n      \"pmids\": [\"17030513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"NMR and crystal structures of human S100A16 homodimer in apo and Ca²⁺-bound forms show that Ca²⁺ binding produces only minor conformational rearrangement (unlike most S100 proteins), attributed to the absence of the glutamate Ca²⁺ ligand in the N-domain EF-hand and to unusually strong hydrophobic interactions between helices 3 and 4 that stabilize the 'closed' form even after Ca²⁺ binding.\",\n      \"method\": \"X-ray crystallography and NMR solution structure determination\",\n      \"journal\": \"Journal of biological inorganic chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — dual structural determination (solid state + solution NMR) in apo and Ca²⁺-bound states with functional interpretation; multiple orthogonal structural methods\",\n      \"pmids\": [\"21046186\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"S100A16 promotes adipogenesis in 3T3-L1 preadipocytes: overexpression markedly enhances adipocyte differentiation and proliferation, while RNAi knockdown inhibits both. S100A16 physically interacts with tumor suppressor p53 (co-immunoprecipitation), and S100A16 overexpression inhibits p53-responsive gene expression while knockdown activates them. Elevated intracellular Ca²⁺ causes nuclear exclusion of S100A16. Overexpression also reduces insulin-stimulated glucose uptake and AKT phosphorylation.\",\n      \"method\": \"3T3-L1 differentiation assay, RNAi knockdown, overexpression, co-immunoprecipitation, Western blot, calcium ionophore treatment\",\n      \"journal\": \"Endocrinology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal functional experiments (OE and KD), direct Co-IP for p53 interaction, quantitative adipogenesis and signaling assays in a focused mechanistic study\",\n      \"pmids\": [\"21266506\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"S100A16 inhibits osteogenesis and stimulates adipogenesis in bone marrow-derived mesenchymal stem cells: S100A16 overexpression increases PPARγ promoter luciferase activity and decreases RUNX2 promoter activity, while knockdown has opposite effects. ERK1/2 phosphorylation is involved in osteogenesis regulation whereas JNK phosphorylation is involved in adipogenesis downstream of S100A16.\",\n      \"method\": \"BM-MSC differentiation assay, Oil Red O and Alizarin Red S staining, promoter luciferase assay, Western blot for ERK1/2 and JNK phosphorylation, transgenic/knockout mouse-derived cells\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — promoter luciferase (direct transcriptional readout), functional differentiation assays, signaling assays; single lab with multiple orthogonal readouts\",\n      \"pmids\": [\"23526364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"S100A14 physically interacts with S100A16 (identified by yeast two-hybrid, confirmed by co-immunoprecipitation and co-immunofluorescence). S100A14 overexpression upregulates S100A16 protein without increasing its mRNA, indicating post-transcriptional regulation, whereas S100A16 overexpression does not reciprocally upregulate S100A14 (unidirectional regulation). S100A14/S100A16 degradation is independent of classical proteasomal and lysosomal pathways.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, double immunofluorescence, cycloheximide chase, retroviral overexpression and knockdown, RT-PCR\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — interaction identified by Y2H and confirmed by Co-IP and co-IF; post-transcriptional regulation shown by mRNA/protein discordance; replicated across multiple cell lines\",\n      \"pmids\": [\"24086685\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"S100A16 overexpression in MCF-7 breast cancer cells upregulates Notch1, ZEB1, and ZEB2 transcription factors, leading to repression of E-cadherin and β-catenin and induction of N-cadherin and vimentin (EMT). Notch1-specific siRNA knockdown reverses EMT induced by S100A16 overexpression, placing Notch1 downstream of S100A16 in this pathway.\",\n      \"method\": \"Overexpression in MCF-7 cells, siRNA knockdown of Notch1, Western blot, migration/invasion assays, colony formation\",\n      \"journal\": \"Journal of biomedical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via siRNA rescue, Western blot pathway analysis; single lab with two complementary experimental approaches\",\n      \"pmids\": [\"25287362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"S100A16 overexpression in DU-145 prostate cancer cells activates AKT and ERK signaling and downregulates p21 and p27. Pharmacological inhibition of AKT (LY294002) or ERK (PD98059) suppresses S100A16-induced clone formation and invasion, placing AKT and ERK downstream of S100A16.\",\n      \"method\": \"Stable overexpression and shRNA knockdown, Western blot, transwell/wound healing assays, pharmacological inhibitors\",\n      \"journal\": \"Tumour biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pathway position confirmed by pharmacological epistasis with two inhibitors; single lab, multiple assays\",\n      \"pmids\": [\"27240591\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Brain microvascular endothelial cell (HBMEC) exosomes transfer S100A16 to SCLC cells, causing S100A16 translocation from cytoplasm to nucleus. Elevated S100A16 prevents loss of mitochondrial membrane potential (Δψm) and enhances resistance to apoptosis. This protective effect depends on prohibitin-1 (PHB-1) in the mitochondrial inner membrane, as PHB-1 siRNA delivery into S100A16-overexpressing cells weakens the protective effect.\",\n      \"method\": \"Exosome isolation by ultracentrifugation, GW4869 inhibitor treatment, Western blot, immunofluorescence, Annexin V/PI apoptosis assay, JC-1 mitochondrial membrane potential assay, PHB-1 siRNA knockdown\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional epistasis via PHB-1 siRNA, multiple readouts (apoptosis, Δψm), exosome transfer mechanism; single lab\",\n      \"pmids\": [\"30183374\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"S100A16 interacts with calmodulin (CaM) and regulates hepatic lipid metabolism via the CaM/CAMKK2/AMPK signaling pathway. S100A16 transgenic mice show more severe fatty liver on HFD, while knockdown mice show attenuated steatosis. Overexpression of S100A16 inhibits degradation of 11β-HSD1, a downstream effector of S100A16-induced adipogenesis.\",\n      \"method\": \"Transgenic and knockout mouse models, HFD feeding, co-immunoprecipitation (S100A16-CaM interaction), Western blot, RNA sequencing, Oil Red O staining\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CaM interaction by Co-IP, in vivo transgenic/KO validation of pathway; single lab with multiple methods\",\n      \"pmids\": [\"31069793\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"S100A16 interacts with myosin-9 (identified by mass spectrometry and confirmed functionally) in response to increased Ca²⁺ and TGF-β stimulation, promoting cytoskeleton (F-actin) reorganization and EMT progression in renal tubular epithelial cells. S100A16 overexpression in HK-2 cells increases N-cadherin and vimentin and decreases E-cadherin, and is associated with renal tubulointerstitial fibrosis in both UUO mouse model and patient biopsies.\",\n      \"method\": \"Mass spectrometry (binding partner identification), immunohistochemistry, UUO mouse model with S100A16 transgenic/heterozygous knockout mice, overexpression and knockdown in HK-2 cells, Western blot, F-actin staining\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-identified interaction with functional validation in cells and in vivo; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"32094322\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"S100A16 induces EMT in PDAC cells and promotes metastasis via upregulation of TWIST1 and activation of the STAT3 signaling pathway, as shown by in vitro and in vivo experiments.\",\n      \"method\": \"siRNA knockdown, overexpression, Western blot, in vivo xenograft, TCGA correlation analysis\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo loss/gain-of-function with pathway marker analysis; single lab\",\n      \"pmids\": [\"33359364\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"S100A16 physically interacts with GRP78 in the endoplasmic reticulum of HK-2 cells (co-immunoprecipitation, immunofluorescence colocalization). S100A16 overexpression causes GRP78 to relocalize from ER to cytoplasm and competitively displaces IRE1α from GRP78, leading to IRE1α phosphorylation and XBP1 splicing (ER stress activation). Calcium chelation with BAPTA-AM blocks both the cytoplasmic colocalization and the upregulation of ER stress markers.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, lentiviral overexpression, BAPTA-AM chelation, Western blot for ER stress markers\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, immunofluorescence colocalization, competitive binding demonstrated, Ca²⁺-dependence validated pharmacologically; multiple orthogonal methods in single focused study\",\n      \"pmids\": [\"34645789\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"S100A16 promotes pancreatic cancer cell proliferation, migration, and invasion via FGF19-dependent activation of AKT and ERK1/2 signaling. S100A16 knockdown induces G2/M cell cycle arrest and apoptosis.\",\n      \"method\": \"Knockdown and overexpression, in vitro proliferation/migration/invasion assays, in vivo metastasis model, Western blot\",\n      \"journal\": \"Cell biology and toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss and gain of function with FGF19 dependency established; single lab with in vitro and in vivo readouts\",\n      \"pmids\": [\"33389337\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"S100A16 promotes gastric cancer cell invasion and EMT via ubiquitination and degradation of ZO-2 (Zonula Occludens-2), a tight junction regulator, identified by proteomic analysis and validated by functional assays.\",\n      \"method\": \"Proteomics/mass spectrometry (interactome), overexpression and knockdown, functional invasion/migration assays, Western blot for ZO-2 ubiquitination\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-identified interaction, ubiquitination-mediated degradation mechanism shown; single lab\",\n      \"pmids\": [\"34650982\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"ADAMTS19 binds cytoplasmic p65 and decreases nuclear phospho-p65 (NF-κB), thereby suppressing S100A16 transcription. S100A16 acts downstream of ADAMTS19 to promote gastric cancer cell migration and invasion, and rescue of S100A16 reverses the suppression caused by ADAMTS19 overexpression.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, dual-luciferase reporter assay, gain/loss-of-function, transwell assay\",\n      \"journal\": \"Biomolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and luciferase confirm NF-κB→S100A16 transcriptional axis; epistasis rescue experiment; single lab\",\n      \"pmids\": [\"33921267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"S100A16 promotes Wnt/β-catenin signaling activation in renal interstitial fibroblasts during acute kidney injury by facilitating HRD1 (E3 ubiquitin ligase)-mediated ubiquitination and degradation of GSK3β and CK1α (negative regulators of β-catenin). S100A16 knockout in mice subjected to ischemia-reperfusion injury impedes Wnt/β-catenin activation and restores HGF expression.\",\n      \"method\": \"S100A16 knockout and overexpression in mice and NRK-49F cells, IRI model, Western blot, ubiquitination assay, ICG-001 Wnt inhibitor\",\n      \"journal\": \"Cellular and molecular life sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo KO and in vitro OE with convergent mechanistic findings; ubiquitination mechanism identified; pathway position confirmed by pharmacological Wnt inhibitor; multiple orthogonal methods\",\n      \"pmids\": [\"35279748\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"S100A16 binds to p53 protein, inducing its degradation; this augments CXCR4 expression, which activates ERK1/2 and AKT signaling, promoting hepatic stellate cell activation and liver fibrosis. S100a16 transgenic mice develop spontaneous liver fibrosis while S100a16 knockout mice are protected.\",\n      \"method\": \"HSC isolation, S100a16 knockout and transgenic mice, multiple fibrosis models, transcriptome sequencing (RNA-seq), co-immunoprecipitation (S100A16-p53), Western blot\",\n      \"journal\": \"Metabolism: clinical and experimental\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo transgenic and KO phenotypes, Co-IP for p53 interaction, RNA-seq for pathway identification, multiple disease models; multiple orthogonal methods\",\n      \"pmids\": [\"35914619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TFAP2B acts as a transcription factor for S100A16 (confirmed by ChIP and luciferase reporter assay). HIF-1α transcriptionally regulates HRD1 (confirmed by ChIP and luciferase reporter assay) within the S100A16→HRD1→GSK3β/CK1α pathway in renal hypoxia injury. S100A16 deletion attenuates HIF-1α upregulation during IRI.\",\n      \"method\": \"ChIP assay, luciferase reporter assay, S100A16 knockout rat cell line (NRK-52E), hypoxia/reoxygenation model, Western blot\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — ChIP and luciferase reporter for transcription factor binding; single lab, two orthogonal methods for each transcriptional relationship\",\n      \"pmids\": [\"38710691\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"During myocardial ischemia/reperfusion injury, VDAC1 upregulation activates NF-κB/p65, which binds to the S100A16 promoter to drive S100A16 transcription. S100A16 then interacts with calmodulin (CaM) in response to elevated Ca²⁺ to activate the CAMKK2/AMPK pathway, contributing to inflammation and ROS production. Adenovirus-mediated S100A16 inhibition improves cardiac function and reduces infarct size.\",\n      \"method\": \"In vivo cardiac I/R model (ligation/release), H/R cell model, adenovirus-mediated S100A16 inhibition, ChIP-like and luciferase assays for NF-κB promoter binding, Western blot, co-immunoprecipitation (S100A16-CaM)\",\n      \"journal\": \"European journal of pharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for CaM interaction, promoter binding confirmed, in vivo and in vitro concordant results; single lab\",\n      \"pmids\": [\"39613175\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"S100A16 binds to MOV10 RNA helicase (co-immunoprecipitation) and positively modulates MOV10 expression in lung adenocarcinoma cells. MOV10 in turn stabilizes ITGA3 mRNA (RNA immunoprecipitation, actinomycin D mRNA stability assay), thereby regulating ECM-receptor interaction signaling and promoting malignant properties. MOV10 overexpression partially reverses the suppressive effects of S100A16 knockdown.\",\n      \"method\": \"Co-immunoprecipitation (S100A16-MOV10), RNA immunoprecipitation (MOV10-ITGA3 mRNA), actinomycin D mRNA stability assay, knockdown and overexpression, Western blot\",\n      \"journal\": \"Molecular medicine reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and RIP with mRNA stability assay establish mechanism; epistasis rescue; single lab with three orthogonal methods\",\n      \"pmids\": [\"39450567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"S100A16 knockdown in HeLa and SiHa cervical cancer cells inhibits cell migration. S100A16 regulates RPN2 (ribophorin II) expression via phospho-STAT3, which in turn modulates phospho-GSK3β to activate β-catenin/TCF signaling. S100A16 silencing reduces nuclear translocation of β-catenin (shown by nuclear/cytosolic fractionation). RPN2 overexpression reverses the migration suppression caused by S100A16 knockdown.\",\n      \"method\": \"siRNA knockdown, RNA sequencing, nuclear/cytosolic fractionation, Western blot, overexpression rescue experiment, wound-healing migration assay\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNA-seq for downstream target, fractionation for β-catenin localization, rescue epistasis; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"40907797\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"S100A16 is enriched in nucleoli of metastatic breast cancer cells (nucleolar proteomics). ChIP-MS shows S100A16 associates at rDNA loci with RPA194 (catalytic subunit of RNA Polymerase I). Loss of S100A16 disrupts RNA Polymerase I activation and rRNA synthesis, reverses EMT, inhibits invasion, and reduces metastatic incidence in animal models.\",\n      \"method\": \"Nucleolar proteomics, ChIP-MS (rDNA loci, RPA194), loss-of-function in vitro and in vivo metastasis assays, rRNA synthesis assay\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — ChIP-MS directly links S100A16 to rDNA/RNA Pol I; nucleolar proteomics for localization; in vitro and in vivo functional validation; multiple orthogonal methods in single study\",\n      \"pmids\": [\"40846689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"S100A14 physically interacts with S100A16 (confirmed by Co-IP), stabilizes S100A16 protein via post-translational modification without transcriptional regulation (CHX chase and mRNA analysis). The S100A14/S100A16 complex reduces p53 protein stability and inhibits p53 transcriptional activity and downstream p21 expression. Knockdown of S100A14 restores p53 function, consistent with the previously described S100A16-p53 interaction.\",\n      \"method\": \"Co-immunoprecipitation, CHX chase assay, dual-luciferase assay (p53 transcriptional activity), siRNA knockdown, Western blot\",\n      \"journal\": \"Oncology research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, protein stability assay, and transcriptional activity assay confirm mechanism; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"41799516\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CRYAB lysine 92 lactylation (CRYAB_K92la) enhances S100A16 transcription, stabilizes S100A16 protein by inhibiting its degradation, and strengthens CRYAB-S100A16 binding. Upregulated S100A16 activates RAGE signaling, which promotes lactate production, forming a glycolysis-fibrosis positive feedback loop in ligamentum flavum hypertrophy.\",\n      \"method\": \"Mass spectrometry (lactylation identification), CRYAB_K92R lactylation-blocking mutation, co-immunoprecipitation (CRYAB-S100A16), promoter assay (S100A16 transcription), Western blot, in vitro fibrosis assays\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — PTM identified by MS, functional mutation (K92R), Co-IP for protein-protein interaction; single lab with multiple methods\",\n      \"pmids\": [\"42098255\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"S100A16 preferentially interacts with saturated phospholipids with short acyl chains and zwitterionic polar head groups, and with cholesterol-enriched membrane domains (lipid raft-like), as determined using Langmuir monolayer biophysical assays. These interactions are calcium-sensitive and suggest S100A16 associates with plasma membrane and nascent disk membranes in photoreceptor outer segments.\",\n      \"method\": \"Langmuir monolayer model, surface tensiometry\",\n      \"journal\": \"Archives of biochemistry and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro reconstitution biophysical assay with defined lipid compositions; single lab, single method\",\n      \"pmids\": [\"42203137\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"S100A16 knockdown in renal cancer cells inhibits proliferation and migration and reduces VEGF, VEGFR2, and phospho-AKT expression, implicating S100A16 in the VEGF/VEGFR2/PI3K-AKT signaling axis in renal cell carcinoma.\",\n      \"method\": \"siRNA knockdown, Western blot, CCK8, wound healing/transwell assays\",\n      \"journal\": \"Contrast media & molecular imaging\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, Western blot for pathway markers after knockdown; no direct binding or reconstitution experiments\",\n      \"pmids\": [\"36176934\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"S100A16 knockdown in bladder cancer cells suppresses the AKT/Bcl-2 pathway to promote apoptosis and sensitizes cells to mitomycin C. The EMT-associated transcription factor Snail regulates S100A16 expression in resistant bladder cancer cells.\",\n      \"method\": \"Proteomics (LC-MS/MS) for initial identification, siRNA knockdown, Western blot, CCK8 viability assay\",\n      \"journal\": \"Cancer management and research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — Snail-S100A16 regulatory link inferred from expression correlation after proteomics; siRNA knockdown with pathway Western blot; no direct binding assay; single lab\",\n      \"pmids\": [\"31118765\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"S100A16 deletion protects mice against alcoholic liver lipid accumulation and inflammation by upregulating mesencephalic astrocyte-derived neurotrophic factor (MANF), which inhibits ER stress signaling. MANF silencing suppressed the inhibitory effect of S100a16 knockout on ethanol-induced lipid droplet accumulation in primary hepatocytes.\",\n      \"method\": \"S100a16 knockout and transgenic mice, Gao-binge alcohol feeding model, MANF siRNA, Western blot, lipid staining, ELISA\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo KO and transgenic with epistasis rescue (MANF siRNA); concordant in vitro and in vivo; single lab\",\n      \"pmids\": [\"37928262\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Co-silencing of S100A16 and HSP27 in placenta-derived multipotent stem cells (PDMCs) without chemical induction drives differentiation into functional astrocytes (confirmed by morphology, neural marker immunofluorescence, electrophysiology, and Ca²⁺ influx measurement), indicating S100A16 suppresses astrocyte differentiation.\",\n      \"method\": \"siRNA knockdown (combinatorial), immunofluorescence quantification, electrophysiology, Ca²⁺ influx assay, transcriptomic/proteomic cross-comparison\",\n      \"journal\": \"Stem cell reviews and reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional differentiation with electrophysiological confirmation; multiple orthogonal validation methods; single lab\",\n      \"pmids\": [\"35061207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"S100A16 knockdown in nephroblastoma cells inhibits proliferation, invasion, migration, and angiogenesis. S100A16 physically interacts with DEPDC1 (co-immunoprecipitation), and DEPDC1 overexpression partially reverses the suppressive effects of S100A16 interference, including suppression of PI3K/Akt/mTOR pathway activation.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, overexpression rescue, CCK8, Transwell, tube formation assay, Western blot\",\n      \"journal\": \"Polish journal of pathology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP, epistasis by overexpression rescue; single lab, limited mechanistic depth\",\n      \"pmids\": [\"37955537\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"In cancer stem-like spheroids of Yumoto cervical carcinoma cells, S100A16 knockdown decreases Oct4 and Nanog mRNA and protein expression and reduces sphere size. S100A16 knockdown also restores p53 protein that was diminished during sphere formation. Proteasome inhibitor lactacystin decreases Oct4 and Nanog protein but does not affect S100A16 protein, indicating S100A16 maintains Oct4/Nanog via a proteasome-dependent mechanism without being itself a proteasomal target.\",\n      \"method\": \"Sphere formation assay, siRNA knockdown, RT-PCR, Western blot, proteasome inhibitor (lactacystin) treatment\",\n      \"journal\": \"Oncology letters\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, indirect mechanistic inference from proteasome inhibitor; limited pathway resolution\",\n      \"pmids\": [\"29928366\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"S100A16 is a homodimeric EF-hand calcium-binding protein that senses Ca²⁺ through its C-terminal EF-hand (the N-terminal site is non-functional), undergoes Ca²⁺-dependent conformational changes and nucleolar-to-cytoplasmic translocation, and executes diverse cellular functions through multiple experimentally confirmed molecular interactions: it binds and inhibits p53 (promoting adipogenesis and cancer progression), interacts with calmodulin to regulate the CaM/CAMKK2/AMPK pathway (controlling lipid metabolism and myocardial stress responses), associates with GRP78 in the ER to competitively release IRE1α and activate ER stress via IRE1α/XBP1, interacts with myosin-9 to drive cytoskeletal reorganization and EMT, promotes HRD1-mediated ubiquitination and degradation of GSK3β and CK1α to activate Wnt/β-catenin signaling, localizes to nucleoli where it associates with RNA Polymerase I at rDNA loci to drive rRNA synthesis and support metastasis, and is transcriptionally regulated by TFAP2B, NF-κB/p65, and BRD4-dependent H3K27 acetylation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"S100A16 is a homodimeric EF-hand calcium-binding protein that senses Ca²⁺ exclusively through the C-terminal EF-hand of each subunit—the N-terminal site being non-functional owing to loss of the conserved glutamate ligand—and translates this into Ca²⁺-dependent conformational change, subcellular redistribution, and target engagement that collectively drive differentiation, metabolic, and pro-metastatic programs [#0, #1]. Unlike most S100 proteins, Ca²⁺ binding produces only a minor structural rearrangement because strong hydrophobic helix 3–4 interactions stabilize the closed conformation, yet human S100A16 still forms a Ca²⁺-induced hydrophobic patch for target recognition [#0, #1], and it accumulates in nucleoli, translocating to the cytoplasm or nucleus upon Ca²⁺ stimulation [#0, #2]. A recurrent mechanistic theme is inhibition of the tumor suppressor p53: S100A16 binds p53 and represses its transcriptional output, promoting adipogenesis and, through p53 degradation and downstream CXCR4/ERK/AKT signaling, hepatic stellate cell activation and liver fibrosis in transgenic and knockout mouse models [#2, #16]. S100A16 controls lipid metabolism and stress responses through calmodulin binding and activation of the CaM/CAMKK2/AMPK axis [#8, #18], and engages the ER chaperone GRP78 to competitively displace IRE1α, triggering IRE1α/XBP1-mediated ER stress in a Ca²⁺-dependent manner [#11]. In cancer and fibrosis it drives epithelial–mesenchymal transition and invasion via multiple effectors—myosin-9-dependent cytoskeletal reorganization [#9], and ubiquitin-mediated turnover of negative regulators, facilitating HRD1-mediated degradation of GSK3β and CK1α to activate Wnt/β-catenin signaling [#15]. In metastatic breast cancer it localizes to nucleoli and associates with RNA Polymerase I (RPA194) at rDNA loci to support rRNA synthesis, EMT, and metastasis [#21]. S100A16 is itself stabilized by its paralog S100A14 through a post-translational, non-transcriptional mechanism, with the S100A14/S100A16 complex amplifying p53 suppression [#4, #22], and its transcription is governed by TFAP2B and NF-κB/p65 [#17, #18].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established S100A16 as a genuine EF-hand Ca²⁺ sensor and defined the structural basis of its binding, answering whether and how this S100 family member responds to calcium.\",\n      \"evidence\": \"Flow dialysis Ca²⁺ binding, Trp fluorescence, and live-cell localization of recombinant human/mouse protein in glioblastoma cells\",\n      \"pmids\": [\"17030513\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not identify physiological Ca²⁺-dependent target proteins\", \"Functional consequence of nucleolar-to-cytoplasmic translocation unknown\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Resolved why S100A16 behaves atypically among S100 proteins by showing Ca²⁺ binding causes only minimal conformational change, refining the structural model.\",\n      \"evidence\": \"X-ray crystallography and NMR solution structures of apo and Ca²⁺-bound homodimer\",\n      \"pmids\": [\"21046186\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of specific target recognition not resolved\", \"No co-structure with a partner protein\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Connected S100A16 to adipogenesis and identified p53 as a direct binding partner whose activity it represses, providing the first molecular partner and a tumor-suppressor link.\",\n      \"evidence\": \"3T3-L1 differentiation with reciprocal overexpression/knockdown, p53 Co-IP, and Ca²⁺-ionophore-induced nuclear exclusion\",\n      \"pmids\": [\"21266506\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Binding interface and Ca²⁺-dependence of p53 interaction not mapped\", \"Mechanism by which binding represses p53 transcriptional output unclear\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Showed S100A16 reciprocally controls the adipogenic/osteogenic fate switch in mesenchymal stem cells through ERK1/2 and JNK signaling, extending its role to lineage commitment.\",\n      \"evidence\": \"BM-MSC differentiation, PPARγ/RUNX2 promoter luciferase, and phospho-MAPK Western blots\",\n      \"pmids\": [\"23526364\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular link between S100A16 and MAPK activation not established\", \"No in vivo skeletal phenotype tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Identified the paralog S100A14 as a direct interactor that post-transcriptionally stabilizes S100A16, revealing a regulatory dimerization mechanism.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, co-IF, and cycloheximide chase across multiple cell lines\",\n      \"pmids\": [\"24086685\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Degradation pathway (non-proteasomal, non-lysosomal) not identified\", \"Functional consequence of the complex not yet defined here\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Placed S100A16 upstream of EMT transcription factors in breast cancer, defining a pro-invasive transcriptional program.\",\n      \"evidence\": \"MCF-7 overexpression with Notch1 siRNA rescue and EMT marker analysis\",\n      \"pmids\": [\"25287362\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which S100A16 upregulates Notch1/ZEB unknown\", \"Single cell line\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Extended S100A16 pro-tumor signaling to prostate cancer via AKT/ERK activation and cell-cycle inhibitor downregulation.\",\n      \"evidence\": \"DU-145 overexpression/knockdown with LY294002 and PD98059 pharmacological epistasis\",\n      \"pmids\": [\"27240591\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct binding partner upstream of AKT/ERK identified\", \"Single cell line\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrated intercellular transfer of S100A16 via exosomes confers apoptosis resistance through mitochondrial PHB-1, introducing a non-cell-autonomous mechanism.\",\n      \"evidence\": \"HBMEC-to-SCLC exosome transfer, JC-1 membrane-potential assay, and PHB-1 siRNA epistasis\",\n      \"pmids\": [\"30183374\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct S100A16-PHB-1 binding not shown\", \"Mechanism of nuclear translocation upon transfer unclear\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified calmodulin as a binding partner linking S100A16 to the CaM/CAMKK2/AMPK pathway controlling hepatic lipid metabolism in vivo.\",\n      \"evidence\": \"S100A16 transgenic/knockout mice on HFD, CaM Co-IP, RNA-seq\",\n      \"pmids\": [\"31069793\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ca²⁺-dependence of CaM interaction not formally tested here\", \"Direct effect on AMPK vs indirect not separated\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified myosin-9 as a Ca²⁺/TGF-β-induced interactor driving cytoskeletal reorganization and EMT in renal fibrosis, linking S100A16 to fibrotic disease.\",\n      \"evidence\": \"Mass spectrometry interactome, UUO mouse model with transgenic/knockout mice, HK-2 functional assays\",\n      \"pmids\": [\"32094322\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Binding interface with myosin-9 not mapped\", \"Whether myosin-9 binding is direct not biochemically proven\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed S100A16 drives PDAC metastasis through TWIST1 and STAT3, generalizing its EMT-promoting role to pancreatic cancer.\",\n      \"evidence\": \"Loss/gain-of-function in vitro and xenograft with TCGA correlation\",\n      \"pmids\": [\"33359364\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct molecular partner linking S100A16 to STAT3\", \"Mechanism of TWIST1 induction unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined a Ca²⁺-dependent ER stress mechanism in which S100A16 binds GRP78 and competitively displaces IRE1α to activate IRE1α/XBP1 signaling.\",\n      \"evidence\": \"Reciprocal Co-IP, IF colocalization, competitive binding, BAPTA-AM Ca²⁺ chelation in HK-2 cells\",\n      \"pmids\": [\"34645789\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry of GRP78 displacement not quantified\", \"Whether mechanism operates outside renal cells untested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Linked S100A16 to FGF19-dependent AKT/ERK signaling driving pancreatic cancer proliferation and survival.\",\n      \"evidence\": \"Knockdown/overexpression, cell-cycle and apoptosis assays, in vivo metastasis model\",\n      \"pmids\": [\"33389337\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism connecting S100A16 to FGF19 unclear\", \"No direct binding partner identified\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified ZO-2 as a degradation target, showing S100A16 promotes gastric cancer invasion by ubiquitin-mediated turnover of a tight-junction regulator.\",\n      \"evidence\": \"Proteomic interactome, functional assays, ZO-2 ubiquitination Western blot\",\n      \"pmids\": [\"34650982\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E3 ligase mediating ZO-2 ubiquitination not identified\", \"Direct vs indirect role of S100A16 in degradation unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Established an NF-κB-driven transcriptional axis upstream of S100A16 suppressed by ADAMTS19 in gastric cancer.\",\n      \"evidence\": \"Co-IP, dual-luciferase reporter, and S100A16 rescue epistasis\",\n      \"pmids\": [\"33921267\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct p65 binding to S100A16 promoter assumed not fully mapped\", \"Single cancer context\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Resolved how S100A16 activates Wnt/β-catenin by facilitating HRD1-mediated ubiquitination and degradation of GSK3β and CK1α, providing a defined enzymatic mechanism in kidney injury.\",\n      \"evidence\": \"S100A16 knockout/overexpression mice and NRK-49F cells, IRI model, ubiquitination assay, ICG-001 Wnt inhibitor\",\n      \"pmids\": [\"35279748\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether S100A16 directly scaffolds HRD1 onto substrates not biochemically shown\", \"Ca²⁺-dependence of this mechanism untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Mechanistically tied S100A16-mediated p53 degradation to CXCR4/ERK/AKT signaling in hepatic stellate cell activation and liver fibrosis with strong in vivo support.\",\n      \"evidence\": \"Transgenic and knockout mice across fibrosis models, p53 Co-IP, RNA-seq\",\n      \"pmids\": [\"35914619\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E3 ligase for p53 degradation not identified\", \"Direct CXCR4 regulation mechanism not mapped\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identified TFAP2B as a direct transcription factor for S100A16 and integrated HIF-1α/HRD1 regulation into the renal hypoxia pathway.\",\n      \"evidence\": \"ChIP and luciferase reporter assays, S100A16 knockout NRK-52E cells, H/R model\",\n      \"pmids\": [\"38710691\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relative contribution of TFAP2B vs NF-κB in different tissues unclear\", \"Single experimental system\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showed VDAC1/NF-κB-driven S100A16 transcription and CaM/CAMKK2/AMPK activation contribute to myocardial ischemia/reperfusion injury, defining a cardiac stress axis.\",\n      \"evidence\": \"Cardiac I/R and H/R models, adenoviral inhibition, promoter binding and CaM Co-IP\",\n      \"pmids\": [\"39613175\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct VDAC1-S100A16 relationship not established\", \"Ca²⁺-dependence of CaM interaction in cardiac context not isolated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed an RNA-regulatory function in which S100A16 binds the MOV10 helicase to stabilize ITGA3 mRNA and promote lung adenocarcinoma malignancy.\",\n      \"evidence\": \"Co-IP, RIP, actinomycin D mRNA stability assay, knockdown/overexpression rescue\",\n      \"pmids\": [\"39450567\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect MOV10 binding not fully resolved\", \"Whether S100A16 itself binds RNA untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined a nucleolar function: S100A16 associates with RNA Polymerase I (RPA194) at rDNA loci to drive rRNA synthesis and support breast cancer metastasis, explaining its long-observed nucleolar localization.\",\n      \"evidence\": \"Nucleolar proteomics, ChIP-MS at rDNA loci, loss-of-function in vitro and in vivo metastasis assays\",\n      \"pmids\": [\"40846689\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How S100A16 is recruited to rDNA not defined\", \"Ca²⁺-dependence of RNA Pol I association untested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Connected S100A16 to RPN2/STAT3/GSK3β-dependent β-catenin nuclear translocation driving cervical cancer migration.\",\n      \"evidence\": \"siRNA knockdown, RNA-seq, nuclear/cytosolic fractionation, RPN2 rescue\",\n      \"pmids\": [\"40907797\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct partner linking S100A16 to RPN2 expression unknown\", \"Single cancer type\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Confirmed the S100A14/S100A16 complex amplifies p53 suppression through post-translational stabilization of S100A16, unifying paralog interaction with p53 regulation.\",\n      \"evidence\": \"Co-IP, CHX chase, dual-luciferase p53 activity assay, S100A14 knockdown\",\n      \"pmids\": [\"41799516\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Identity of the stabilizing post-translational modification unknown\", \"Stoichiometry of the complex on p53 not defined\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed CRYAB K92 lactylation enhances S100A16 transcription and stability and strengthens CRYAB-S100A16 binding, embedding S100A16 in a glycolysis-fibrosis RAGE feedback loop.\",\n      \"evidence\": \"Mass spectrometry, K92R lactylation-blocking mutant, Co-IP, promoter assay, fibrosis assays\",\n      \"pmids\": [\"42098255\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct RAGE binding by S100A16 not shown\", \"Single disease context\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Characterized Ca²⁺-sensitive S100A16 interactions with saturated short-chain phospholipids and cholesterol-rich membrane domains, suggesting a membrane-association function.\",\n      \"evidence\": \"Langmuir monolayer biophysical assays with defined lipid compositions\",\n      \"pmids\": [\"42203137\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single in vitro biophysical method\", \"Membrane association not validated in cells\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single small Ca²⁺ sensor with minimal Ca²⁺-induced conformational change selects among its many reported partners (p53, CaM, GRP78, myosin-9, MOV10, RNA Pol I) in different cell types, and whether a unifying biochemical rule governs target choice, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No co-structure of S100A16 with any partner\", \"Ca²⁺-dependence not uniformly tested across interactions\", \"Tissue-specific determinants of partner selection unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005509\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [13, 15]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 11, 16]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [9]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [24]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [0, 21]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [11]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 7]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [24]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [8, 15, 18]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [11, 27]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [21]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [16, 9]}\n    ],\n    \"complexes\": [\"S100A14/S100A16 complex\"],\n    \"partners\": [\"S100A14\", \"TP53\", \"CALM1\", \"HSPA5\", \"MYH9\", \"MOV10\", \"CRYAB\", \"DEPDC1\"],\n    \"other_free_text\": []\n  }\n}\n```\n\nNote: I included GO:0005509 (calcium ion binding) for molecular_activity as it is the central, High-confidence biochemical activity directly demonstrated in [#0] and [#1]; if the controlled vocabulary restricts to the listed slim terms only, this entry should be dropped, but the EF-hand Ca²⁺-binding activity is the defining function of this protein and is strongly evidenced.","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}