| 2003 |
Munc13-4 (encoded by UNC13D/FHL3 locus) is essential for the priming step of cytolytic granule secretion preceding vesicle membrane fusion in cytotoxic T lymphocytes. Mutations in hMunc13-4 cause FHL3. hMunc13-4 deficiency results in defective cytolytic granule exocytosis despite normal granule polarization and docking. Tagged hMunc13-4 localizes with cytotoxic granules at the immunological synapse. |
Patient mutation analysis, immunofluorescence localization of tagged protein, functional cytotoxicity assays in patient cells |
Cell |
High |
14622600
|
| 2003 |
FHL3 (the LIM-domain protein) binds actin directly (confirmed by GST pull-down and co-immunoprecipitation of recombinant and endogenous proteins), localizes to actin stress fibers and focal adhesions following integrin engagement, and inhibits alpha-actinin-mediated actin bundling as demonstrated by low-speed co-sedimentation assays and electron microscopy, thereby promoting stress fiber disassembly and cell spreading. |
Yeast two-hybrid, GST pull-down, co-immunoprecipitation, immunofluorescence, low-speed actin co-sedimentation assay, electron microscopy, overexpression/wound healing assay |
The Journal of biological chemistry |
High |
12704194
|
| 2004 |
FHL3 binds both the alpha-7 and beta-1 subunits of the muscle alpha7beta1 integrin receptor. The suprastructure of the whole FHL3 molecule (not any single LIM domain) is required for integrin binding. FHL3 colocalizes with alpha7beta1 integrin at the periphery of Z-discs in skeletal muscle, suggesting a role in mechanical stabilization. |
Co-immunoprecipitation, deletion mapping, immunofluorescence colocalization |
The Journal of biological chemistry |
Medium |
15117962
|
| 2001 |
FHL3 physically interacts with FHL2; LIM2 of FHL3 is the essential domain for this interaction. LIM1 of FHL3 is essential for its subcellular localization to focal adhesions. FHL2 and FHL3 colocalize in mitochondria of C2C12 cells as demonstrated by GFP-FRET. |
Yeast two-hybrid, site-directed mutagenesis, GFP-fusion FRET in C2C12 cells |
Journal of cellular biochemistry |
Medium |
11135358
|
| 2003 |
FHL3 interacts with the transcriptional repressor BKLF/KLF3 and its co-repressor CtBP2, forming a trimeric complex demonstrated by gel filtration co-elution. BKLF expression promotes nuclear accumulation of FHL3 and CtBP2. FHL3 can repress transcription in this context, contrary to its usual co-activator role. |
Co-immunoprecipitation, gel filtration co-elution, transcriptional reporter assays, immunofluorescence |
The Journal of biological chemistry |
Medium |
12556451
|
| 2003 |
FHL3 interacts with CDC25B2 phosphatase specifically (not other CDC25B isoforms); the second LIM domain of FHL3 is essential for this interaction. FRET in C2C12 cells shows the interaction occurs in the nucleus. FHL3 binding does not impair CDC25B2 phosphatase activity or localization. |
Yeast two-hybrid, deletion/point mutation analysis, FRET in C2C12 cells, phosphatase activity assay |
Experimental cell research |
Medium |
12681290
|
| 2007 |
FHL3 binds MyoD directly (demonstrated by direct binding assays, nuclear colocalization, and GST pull-down) and functions as a potent negative co-transcriptional regulator of MyoD. FHL3 overexpression retards myotube formation and decreases myogenin (but not MyoD) expression; siRNA-mediated FHL3 knockdown enhances differentiation and MyoD transcriptional activity in a dose-dependent manner. |
GST pull-down, co-immunoprecipitation, immunofluorescence colocalization, siRNA knockdown, overexpression, transcriptional reporter assays |
Journal of cell science |
High |
17389685
|
| 2007 |
Fhl3 is recruited by Sox15 to transcriptionally coactivate Foxk1 gene expression in myogenic progenitor cells. Sox15 binds an evolutionarily conserved site in the Foxk1 promoter and recruits Fhl3; knockdown of Sox15 decreases Foxk1 expression and perturbs cell cycle kinetics. |
Transgenic mouse reporter assay, chromatin binding assay, siRNA knockdown, Sox15 mutant mice with satellite cell counting |
The EMBO journal |
Medium |
17363903
|
| 2013 |
PCBP2 binds FHL3 mRNA (identified by RIP-ChIP) and destabilizes it, suppressing FHL3 protein expression. Knockdown of PCBP2 enhances FHL3 mRNA stability and increases FHL3 protein levels. FHL3 overexpression attenuates glioma cell growth and induces apoptosis. |
RIP-ChIP (RNA immunoprecipitation + microarray), siRNA knockdown, overexpression, in vitro and in vivo tumor growth assays |
The Journal of clinical investigation |
Medium |
23585479
|
| 2015 |
FHL3 differentially regulates MyHC isoform expression: it downregulates MyHC 1/slow via inhibiting MyoD transcriptional activity, and upregulates MyHC 2a via interaction with phosphorylated CREB (pCREB). FHL3 interacts with MyoD, CREB, and pCREB in vivo; pCREB shows stronger binding to CRE of the MyHC 2a promoter than CREB, and FHL3 significantly affects pCREB binding capacity to CRE. |
Co-immunoprecipitation, chromatin immunoprecipitation (ChIP), transcriptional reporter assays, overexpression |
Cellular signalling |
Medium |
26499038
|
| 2018 |
FHL3 interacts with the Smad2/3 protein complex at the SOX4 promoter region in glioma stem cells (GSCs), recruits PPM1A phosphatase to Smad2/3, inhibits SOX4 transcriptional activity, and suppresses GSC tumor sphere formation and self-renewal via downregulation of SOX2. |
Co-immunoprecipitation, ChIP, transcriptional reporter assays, sphere formation assay, in vivo tumor assays, siRNA knockdown |
Cell death and differentiation |
Medium |
29955125
|
| 2020 |
FHL3 directly binds GSK3β (the LIM-3 domain of FHL3 is required); this interaction weakens the association between GSK3β and Snail1/Twist1, thereby inhibiting ubiquitin-proteasome-mediated degradation of these EMT transcription factors and promoting pancreatic cancer invasion and metastasis. |
Co-immunoprecipitation, domain deletion mapping, GSK3β inhibitor experiments, in vitro and in vivo invasion assays |
Aging |
Medium |
31935687
|
| 2019 |
PCBP2 inhibits FHL3 expression by binding to the FHL3 3'UTR; PCBP2 activates TGF-β/Smad signaling by suppressing FHL3, and this pathway promotes glioma development. Knockdown of FHL3 reverses the antitumor effect of PCBP2 silencing. |
3'UTR binding assay, siRNA knockdown, Western blot for pathway proteins, in vivo tumor assay |
Journal of cellular physiology |
Medium |
31693182
|
| 2021 |
FHL3 competitively binds the ubiquitin complex (Slug/GSK3β/RNF146), preventing ubiquitination of Slug and up-regulating Slug expression, thereby promoting EMT and chemotherapy resistance in gastric cancer via MAPK/ERK and PI3K/Akt/GSK3β pathways. |
Co-immunoprecipitation, siRNA knockdown, overexpression, in vivo tumor xenograft, Western blot for pathway components |
Frontiers in oncology |
Medium |
34150617
|
| 2021 |
Fhl3 (Xenopus ortholog) interacts with Smad1 and promotes Smad1 binding to the wnt8 promoter in a BMP-dependent manner during neural crest induction, thereby enhancing BMP signaling output and coordinating BMP-WNT signaling in neural crest development. |
Co-immunoprecipitation, ChIP, loss-of-function (morpholino), gain-of-function, in vivo Xenopus embryo assays |
Cell reports |
Medium |
34161771
|
| 2012 |
FHL3 is a novel angiogenin (Ang)-binding partner identified by yeast two-hybrid screening and confirmed by co-immunoprecipitation and GST pull-down. FHL3 is required for Ang-mediated HeLa cell proliferation and nuclear translocation of Ang. |
Yeast two-hybrid, co-immunoprecipitation, GST pull-down, siRNA knockdown, cell proliferation assay |
Gene |
Medium |
22633874
|
| 2010 |
The second zinc finger motif within LIM4 of FHL3 is responsible for its transactivation activity, as determined by sequential LIM domain deletion mutants assayed in a yeast auto-activation system. |
Yeast transcriptional activation assay, site-directed mutagenesis, Western blot |
Molekuliarnaia biologiia |
Low |
20586194
|
| 2023 |
FHL3 directly interacts with the YY1 DNA-binding domain, represses YY1 binding to the MyHC2b gene regulatory region, and competes with EZH2 for binding to YY1's repression domain, thereby reducing H3K27me3 enrichment at the MyHC2b regulatory region and promoting fast glycolytic muscle fiber formation. In vivo, muscle-specific FHL3 overexpression increases fast-twitch myofiber proportion and reduces SDH activity. |
Co-immunoprecipitation, ChIP, transgenic mice, lentivirus-mediated overexpression/knockdown in mice and pigs |
Cellular and molecular life sciences |
High |
36602641
|
| 2025 |
FHL3 interacts with MAZ (MYC-associated zinc finger protein) to recruit MAZ to G-quadruplex structures in the KRAS promoter, promoting KRAS transcription and downstream oncogenic signaling in hepatocellular carcinoma. YAP transcriptionally activates FHL3 as demonstrated by luciferase reporter assay and ChIP. |
Co-immunoprecipitation, ChIP, luciferase reporter assay, siRNA knockdown, in vivo tumor model |
Cell death & disease |
Medium |
41184244
|
| 2025 |
AMBP competitively binds the zinc finger domain of FHL3, disrupting FHL3's protective role in preventing ubiquitin-proteasome-mediated degradation of phospho-ERK1/2 and phospho-JNK, thereby inhibiting osteoblastic differentiation of valvular interstitial cells and aortic valve calcification. |
Co-immunoprecipitation, AlphaFold3 structural modeling, siRNA knockdown, pharmacological inhibitors/agonists, in vivo mouse model |
Theranostics |
Medium |
40225558
|
| 2025 |
FHL3 localizes to mitochondria-lipid droplet contact sites in HepG2 cells; suppression of FHL3 leads to reduced lipid droplet size, elongated mitochondrial morphology, and diminished mitochondria-lipid droplet interactions, suggesting a role in regulating fatty acid β-oxidation. |
Microscopy-guided spatial proteomics (Microscoop Mint), immunofluorescence, siRNA knockdown with organelle morphology readouts |
bioRxivpreprint |
Low |
bio_10.1101_2025.01.24.626799
|
| 2014 |
MT-1X (Metallothionein-1X) is a novel FHL3 binding partner identified by yeast two-hybrid and confirmed by co-immunoprecipitation and GST pull-down. MT-1X knockdown promotes FHL3-induced inhibitory effects on HepG2 cells by regulating FHL3-mediated Smad signaling and G2/M phase-related protein expression. |
Yeast two-hybrid, co-immunoprecipitation, GST pull-down, siRNA knockdown, Western blot |
PloS one |
Low |
24690879
|