| 1997 |
14-3-3 proteins (including the beta isoform, YWHAB) bind phosphoserine-containing motifs (RSXpSXP and RXY/FXpSXP) present in nearly all known 14-3-3 binding partners. Crystal structure of 14-3-3zeta complexed with a phosphoserine motif revealed the bound peptide in an extended conformation with a cis-Pro at pS+2, and showed that the 14-3-3 dimer binds tightly to molecules with tandem phosphoserine motifs (bidentate association), implicating this as a signaling mechanism for partners such as Raf, BAD, and Cbl. |
Phosphoserine-oriented peptide library screening of all mammalian and yeast 14-3-3 isoforms; X-ray crystallography (2.6 Å resolution) of 14-3-3zeta/phosphopeptide complex; in vitro binding assays |
Cell |
High |
9428519
|
| 1997 |
14-3-3 proteins (including beta/YWHAB) bind Raf-1 in a phosphorylation-dependent manner and are required for Raf-1 activation, placing 14-3-3 as an essential component of the Raf-1 activation mechanism in response to signaling events. |
Co-immunoprecipitation, functional kinase assays, phosphorylation site analysis reviewed across multiple studies |
Current opinion in cell biology |
Medium |
9069260
|
| 1999 |
14-3-3 proteins (including YWHAB) bind phosphorylated BAD in the cytosol, preventing its translocation to mitochondria. Ca2+-activated calcineurin dephosphorylates BAD, causing its dissociation from 14-3-3 and translocation to mitochondria where it heterodimerizes with Bcl-xL and promotes apoptosis. |
Co-immunoprecipitation, subcellular fractionation, dominant-negative calcineurin expression, pharmacological calcineurin inhibitors, hippocampal neuron imaging |
Science |
High |
10195903
|
| 2000 |
14-3-3 proteins (including YWHAB) interact with HDAC4 and HDAC5 at three phosphorylation sites, sequestering these class II HDACs in the cytoplasm. Loss of 14-3-3 interaction allows HDAC4/5 to translocate to the nucleus, interact with HDAC3, and repress gene expression, thus controlling transcriptional activity through regulated nuclear-cytoplasmic shuttling. |
Co-immunoprecipitation, subcellular localization studies, transcriptional reporter assays, phosphorylation-site mutagenesis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
10869435
|
| 2004 |
14-3-3 proteins (including YWHAB) sequester the CREB coactivator TORC2 in the cytoplasm via a phosphorylation-dependent interaction under resting conditions. Calcium and cAMP signals promote dephosphorylation of TORC2 via calcineurin and inhibition of SIK2 kinase, disrupting TORC2:14-3-3 complexes and allowing TORC2 nuclear entry and CREB-dependent gene activation, establishing 14-3-3 as a cytoplasmic anchor in a calcium/cAMP coincidence detector module. |
Co-immunoprecipitation, subcellular fractionation, luciferase reporter assays, dominant-negative and pharmacological inhibitor studies |
Cell |
High |
15454081
|
| 2007 |
14-3-3 proteins (including YWHAB) bind phosphorylated YAP (at Ser127 by Lats kinase) and mediate its cytoplasmic sequestration, thereby inactivating the YAP oncoprotein. This 14-3-3-dependent cytoplasmic retention is the key mechanism by which the Hippo pathway suppresses YAP transcriptional activity in response to cell contact inhibition. |
Co-immunoprecipitation, phosphorylation assays, subcellular fractionation/immunofluorescence, Lats kinase loss-of-function, YAP mutational analysis in vivo (Drosophila and mammalian cells) |
Genes & development |
High |
17974916
|
| 2008 |
14-3-3 proteins (including YWHAB) bind TSC2 under growth factor conditions, inhibiting the TSC1/2 complex (a negative regulator of mTORC1). Under hypoxia, REDD1 competitively binds 14-3-3, displacing TSC2 and freeing it to suppress mTORC1 activity. REDD1 mutants unable to bind 14-3-3, and TSC2 mutants unable to bind 14-3-3, abrogate hypoxia-dependent mTORC1 regulation. |
Co-immunoprecipitation, site-directed mutagenesis of 14-3-3 binding sites, REDD1 knockout cells, in vitro competition assays, mouse tumor model |
Genes & development |
High |
18198340
|
| 2010 |
14-3-3 proteins (including YWHAB) bind phospho-Ser127 of YAP; additionally Lats phosphorylates YAP at Ser381, which primes subsequent CK1δ/ε phosphorylation to create a phosphodegron that recruits SCF(β-TRCP) E3 ubiquitin ligase, leading to YAP ubiquitination and degradation. Thus 14-3-3 mediates spatial regulation (cytoplasmic retention) while the phosphodegron mediates temporal regulation (degradation) to coordinately suppress YAP oncogenic activity. |
Co-immunoprecipitation, ubiquitination assays, phosphorylation site mutagenesis, cycloheximide chase, mass spectrometry |
Genes & development |
High |
20048001
|
| 2011 |
14-3-3 proteins (including YWHAB) bind AKT-phosphorylated FoxO transcription factors, sequestering them in the cytoplasm and preventing FoxO-dependent transcription of pro-apoptotic and cell-cycle arrest genes. This interaction forms a key node in insulin/IGF-1 signaling downstream of AKT. |
Co-immunoprecipitation, subcellular fractionation, phosphorylation assays, AKT mutant studies, transcriptional reporter assays; reviewed from multiple independent studies |
Biochimica et biophysica acta |
High |
21708191
|
| 2020 |
YWHAB directly interacts with the PCV2 viral protein ORF5, as confirmed by protein-protein interaction assays. YWHAB expression is induced upon ORF5 overexpression and PCV2 infection; YWHAB overexpression strongly inhibits PCV2 replication and suppresses PCV2-induced ER stress, autophagy, ROS production, and apoptosis, while YWHAB knockdown exacerbates these cellular responses. |
Co-immunoprecipitation, protein-protein interaction assays, ectopic overexpression and gene knockdown (siRNA), viral replication assays, ER stress markers, autophagy flux assays, ROS measurement, flow cytometry (apoptosis) |
Veterinary microbiology |
Medium |
33096469
|
| 2021 |
YWHAB forms a phosphorylation-dependent complex with the glucagon receptor (GCGR) and directly interacts with FOXO1 to inhibit glucagon-mediated hepatic glucose production. Ywhab knockout mice show impaired blood glucose homeostasis under pyruvate stimulation, and deletion in primary hepatocytes increases gluconeogenic output. YWHAB thus acts as an inhibitor of glucagon-induced hepatic gluconeogenesis. |
Affinity purification/mass spectrometry (initial identification), co-immunoprecipitation, Ywhab knockout mice, primary hepatocyte deletion, glucose production assays, pyruvate tolerance tests |
FEBS letters |
High |
33641163
|
| 2021 |
In cervical cancer cells, circ-SMARCA5 binds SND1 protein (verified by RNA pulldown), preventing SND1 from interacting with YWHAB (verified by co-immunoprecipitation). This circ-SMARCA5–SND1 interaction thereby downregulates YWHAB activity, and knockdown of SND1 or YWHAB reverses the pro-proliferative/pro-invasive effects of circ-SMARCA5 siRNA, establishing YWHAB as a downstream effector of the circ-SMARCA5/SND1 axis in cervical cancer. |
RNA pulldown assay (circ-SMARCA5 with SND1), co-immunoprecipitation (SND1–YWHAB), siRNA knockdown of SND1/YWHAB, CCK-8, Transwell invasion, Annexin V apoptosis assays, in vivo xenograft |
Cell transplantation |
Medium |
33588586
|
| 2023 |
YWHAB binds PIK3R2 (PI3K regulatory subunit 2) in colon cancer cells, as demonstrated by co-immunoprecipitation. YWHAB knockdown decreases PIK3R2 expression and reduces PI3K/AKT pathway activation (decreased p-PI3K and p-AKT levels), causing G0/G1 cell cycle arrest and apoptosis. PIK3R2 overexpression rescues the effects of YWHAB knockdown, placing YWHAB upstream of PIK3R2 in activating the PI3K/AKT pathway. |
Co-immunoprecipitation (YWHAB–PIK3R2), siRNA knockdown, overexpression plasmid rescue, Western blotting (PI3K/AKT pathway), flow cytometry (cell cycle, apoptosis), CCK-8, TUNEL assay |
Experimental and therapeutic medicine |
Medium |
37090079
|
| 2024 |
The deubiquitinase OTUB1 directly interacts with YWHAB and deubiquitinates it through a catalytic mechanism, thereby stabilizing YWHAB protein levels. OTUB1-mediated YWHAB stabilization in turn suppresses hepatic gluconeogenesis. Deletion of hepatic OTUB1 reduces YWHAB levels, elevates fasting blood glucose, and increases gluconeogenic gene expression; OTUB1 overexpression mitigates diabetic hyperglycemia. |
Co-immunoprecipitation (OTUB1–YWHAB), in vitro deubiquitination assay, hepatic OTUB1 knockout mice (db/db model), hepatocyte overexpression, glucose tolerance tests, gluconeogenic gene expression analysis |
Cellular signalling |
High |
39270917
|
| 2024 |
YWHAB overexpression inhibits migration and invasion of MDA-MB-231 breast cancer cells without affecting proliferation, while YWHAB knockdown promotes migration and invasion of MCF7 cells. Transcriptomic analysis showed YWHAB overexpression downregulates vimentin (a mesenchymal marker), suggesting YWHAB suppresses epithelial-to-mesenchymal properties. The transcription factor IRX5 regulates YWHAB expression by binding its promoter region, as demonstrated by dual luciferase assay. |
Overexpression and siRNA knockdown, migration/invasion assays (Transwell), transcriptomics (RNA-seq), vimentin Western blot, dual luciferase promoter assay (IRX5 regulation) |
Oncology letters |
Medium |
39119237
|
| 2025 |
The small molecule Kuwanon A (KA) directly targets YWHAB and mediates dimer dissociation of the YWHAB dimer, thereby inhibiting the MAPK (Raf/MEK/ERK) signaling pathway, causing cell cycle arrest and inhibiting migration/invasion of hepatocellular carcinoma cells both in vitro and in vivo. Combination with sorafenib showed synergistic anti-tumor effects. |
Target identification (direct binding assays), dimer dissociation assays, MAPK pathway Western blotting, cell cycle assays, migration/invasion assays, in vivo xenograft mouse model |
Cells |
Medium |
41090717
|