| 2005 |
RSK4 is a predominantly cytosolic protein that is constitutively (maximally) activated under serum-starved conditions in all cell types analyzed, unlike RSK1-3 which require growth factor stimulation. Constitutive activation results from constitutive phosphorylation of Ser232, Ser372, and Ser389. Low basal ERK activity in serum-starved cells is sufficient for ~50% of constitutive RSK4 activity. PDK1 gene deletion in mouse embryonic stem cells suggested PDK1 is not required for phosphorylation of the key activation-loop site Ser232, unlike other RSK family members. |
In vitro kinase assays, phospho-specific antibodies, subcellular fractionation, PDK1-knockout mouse embryonic stem cells, immunoblotting |
The Journal of biological chemistry |
High |
15632195
|
| 2004 |
Rsk4 functions as an inhibitor of fibroblast growth factor-RAS-ERK (RTK) signaling. RTK inhibition is specific to RSK4 among RSK family members and depends on a region of RSK4 that is divergent from other RSK members. Rsk4 inhibits transcriptional activation of specific RTK signaling targets as well as ERK activation. Rsk4 expression in extraembryonic tissue is inversely correlated with activated ERK1/2 levels. |
Overexpression and domain-deletion experiments in cell lines, transcriptional reporter assays, ERK activation assays, in situ hybridization/immunostaining for expression vs. pERK correlation |
Molecular and cellular biology |
Medium |
15121846
|
| 2009 |
RSK4 overexpression induces cell cycle arrest and senescence features in normal fibroblasts (IMR90) and malignant colon carcinoma cells (HCT116). RSK4 mRNA levels increase during both replicative and stress-induced senescence. RSK4-induced senescence is mediated through p21, but is independent of p16, p38 MAPKs, and reactive oxygen species. Cells with RSK4 shRNA knockdown partially escape senescence and show increased resistance to cisplatin. E1A expression or Rb siRNA confers resistance to RSK4-mediated senescence. |
RSK4 cDNA overexpression and shRNA knockdown in IMR90 and HCT116 cells; senescence assays; pharmacological inhibitors of p16, p38, ROS; E1A overexpression; Rb siRNA epistasis |
Clinical cancer research |
Medium |
19584160
|
| 2011 |
RSK4 inhibition via shRNA allows normal human fibroblasts to bypass stress-induced senescence (oxidative stress, DNA damage) and oncogene-induced senescence (KRAS-V12, BRAF-E600 overexpression). RSK4-induced senescence is mediated through p21 but is independent of p16, p38 MAPKs, and reactive oxygen species induction. |
shRNA knockdown of RSK4 in normal human fibroblasts; oncogene overexpression; pharmacological inhibitors; senescence assays; p21 inhibition epistasis |
Carcinogenesis |
Medium |
21239520
|
| 2004 |
RSK4 (and PAK5) are direct transcriptional targets of HNF4alpha. HNF4alpha binds to novel sites in the RSK4 promoter as confirmed by chromatin immunoprecipitation (ChIP) cloning and electrophoretic mobility shift assays (EMSA). RSK4 expression is repressed in streptozotocin-induced diabetic rat kidney and brain coincident with HNF4alpha repression. |
ChIP cloning, EMSA with oligonucleotides harboring HNF4alpha binding sites, quantitative RT-PCR in diabetic rat tissue |
Molecular pharmacology |
Medium |
15615695
|
| 2021 |
RSK4 promotes drug resistance and metastasis in lung and bladder cancer. CRISPR and RNAi silencing of RSK4 sensitized tumor cells to chemotherapy and hindered metastasis in vitro and in vivo. X-ray crystallography identified an allosteric binding site on RSK4 through which floxacin antibiotics (e.g., trovafloxacin) block RSK4 kinase activation by mimicking a kinase autoinhibitory mechanism involving RSK4's hydrophobic motif. Deuterium exchange and Markov transient analyses confirmed the allosteric mechanism. |
CRISPR knockdown, RNAi, X-ray crystallography, hydrogen-deuterium exchange, Markov transient analysis, xenograft models, genetically engineered mouse models, bladder tumor explants |
Science translational medicine |
High |
34261798
|
| 2016 |
Tandem affinity purification coupled with nano LC-MS/MS identified 82 RSK4-associated proteins and 137 RSK4m (alternative splice variant)-associated proteins in MDA-MB-231 cells. Bioinformatics analysis indicated enrichment in cell senescence pathways. RSK4 and RSK4m have distinct interactomes, with RSK4m associated with more molecular functions. |
SF-TAP (tandem affinity purification with STREP II + Flag tags) coupled to nano LC-MS/MS, bioinformatics (GO, IPA) |
International journal of biological macromolecules |
Low |
27986634
|
| 2022 |
FOXP2 transcription factor directly binds to the RSK4 (RPS6KA6) promoter and transcriptionally activates its expression. Knockdown of RSK4 reverses the anti-proliferative and pro-apoptotic effects of FOXP2 overexpression in thyroid cancer cells, placing RSK4 downstream of FOXP2. FOXP2/RSK4 regulation is associated with the PI3K/AKT pathway. |
Dual-luciferase reporter assay, chromatin immunoprecipitation (ChIP), JASPAR binding site prediction, rescue/knockdown experiments in TPC-1 cells |
Experimental and therapeutic medicine |
Medium |
35607372
|
| 2019 |
17β-estradiol (E2) reduces RSK4 expression in ER+ breast cancer cells by promoting DNMT3B-mediated methylation of the RSK4 promoter, thereby decreasing RSK4 transcriptional activity. E2-induced RSK4 silencing is associated with increased cell proliferation, clone formation, and invasion. |
Bisulfite genomic sequencing for methylation, luciferase reporter assay for transcriptional activity, Western blotting for DNMT1/3A/3B, MTT/Transwell/flow cytometry functional assays, RT-PCR |
OncoTargets and therapy |
Medium |
31413588
|
| 2023 |
TRAF4 negatively regulates RSK4; TRAF4 knockdown leads to increased RSK4 expression in ovarian cancer cells. USP7 deubiquitinates and stabilizes TRAF4 via co-immunoprecipitation-confirmed interaction, and TRAF4 destabilization leads to RSK4 upregulation and suppression of the PI3K/AKT pathway. This USP7/TRAF4/RSK4/PI3K/AKT axis was confirmed in a xenograft mouse model. |
Co-immunoprecipitation for TRAF4 ubiquitination, siRNA knockdown of USP7 and TRAF4, RSK4 overexpression, Western blotting, xenograft mouse model |
Journal of cancer research and therapeutics |
Medium |
37006049
|
| 2023 |
RSK4 promotes macrophage recruitment and M2 polarization in esophageal squamous cell carcinoma (ESCC) by enhancing secretion of soluble ICAM-1 (sICAM-1) via direct and indirect STAT3 phosphorylation. RSK4-conditioned macrophages then secrete CCL22 to enhance tumor proliferation, migration, and invasion. |
In vitro co-culture experiments, ELISA for sICAM-1 and CCL22, phospho-STAT3 detection, RSK4 overexpression/knockdown in ESCC cells, xenograft mouse model, IHC in human ESCC tissues |
Biochimica et biophysica acta. Molecular basis of disease |
Medium |
38142759
|
| 2026 |
RSK4 stability is regulated by antagonistic post-translational modifications: GSK3β phosphorylates RSK4 at Thr402/Ser406, promoting its proteasomal degradation via the FBXW7-dependent ubiquitin-proteasome pathway. OGT-mediated O-GlcNAcylation of RSK4 at Thr405 antagonizes GSK3β phosphorylation, thereby stabilizing RSK4 and enhancing cancer stem cell properties and radioresistance in ESCC. Inhibition of O-GlcNAcylation with OSMI-4 destabilizes RSK4 and sensitizes ESCC to radiotherapy. |
Site-directed mutagenesis at Thr402/Ser406/Thr405, co-immunoprecipitation with GSK3β and FBXW7, O-GlcNAc detection, proteasome inhibitor experiments, OSMI-4 treatment, patient-derived xenograft and organoid models |
International journal of biological sciences |
High |
41800247
|
| 2024 |
Deletion of exon 7 of Rps6ka6 (creating a severely truncated RSK4 protein) in H2Kb myogenic cells did not affect myoblast proliferation or migration, but significantly reduced myoblast differentiation indices (myosin expression, myosin-expressing cell size, and fusion index). Genetic analysis in CFW mice linked the Rps6ka6 locus to muscle fiber number in fast-twitch extensor digitorum longus muscle. |
CRISPR-Cas9 exon deletion in myogenic cells, myoblast proliferation/migration/differentiation assays, myosin immunostaining, fusion index quantification, CFW mouse genetic analysis with histology |
G3 (Bethesda, Md.) |
Medium |
38577978
|
| 2018 |
TRAF4 knockdown in breast cancer (MDA-MB-231) xenografts led to significant upregulation of RSK4 mRNA and protein, and coincident reduction of P-AKT, P-NF-κB, TGF-β1, TNF-α, MMP2, and MMP9. This places RSK4 as negatively regulated by TRAF4, upstream of the AKT/NF-κB pathway. |
Lentiviral TRAF4-shRNA in MDA-MB-231 cells, xenograft nude mouse model, IHC, Western blot, fluorescence quantitative RT-PCR |
Biochemical and biophysical research communications |
Low |
29684350
|
| 2020 |
RSK4 overexpression in ccRCC cell lines enhances invasive and migratory abilities through upregulation of CD44 and MMP-9 expression; conversely, RSK4 knockdown reduces these abilities and CD44/MMP-9 expression. |
Ectopic RSK4 overexpression and knockdown in ccRCC cell lines, invasion/migration assays, IHC in patient samples, Western blot |
Diagnostic pathology |
Low |
32209138
|
| 2020 |
miR-548d-3p directly targets RSK4 and negatively regulates its expression. Dual-luciferase reporter assay confirmed binding of miR-548d-3p to the RSK4 3'UTR at a predicted interaction site (mutation of this site abolished the effect). miR-548d-3p overexpression phenocopies RSK4 knockdown in gastric cancer cells: increased proliferation, migration, invasion, and decreased apoptosis. |
Dual-luciferase reporter assay with wild-type and mutant RSK4 3'UTR, RT-PCR, Western blot, functional cell assays, nude mouse tumorigenesis |
Cancer management and research |
Medium |
33380838
|
| 2023 |
RSK4 confers paclitaxel resistance in ovarian cancer cells. RSK4 elevation is associated with accelerated phosphorylation of the downstream translation factor eIF4B. RSK inhibitor (BI-D1870) co-treatment restored paclitaxel sensitivity. External RSK4 expression increased cell viability under paclitaxel treatment. |
DNA microarray, qRT-PCR, Western blotting, RSK inhibitor pharmacological treatment, RSK4 overexpression, cell viability assays |
Biochemical and biophysical research communications |
Low |
37660640
|
| 2025 |
RSK4 promotes metastasis and angiogenesis in clear cell renal cell carcinoma (ccRCC) by activating RUNX1-mediated transcription of EPHA2, leading to enhanced VEGF secretion and tube formation. Luciferase reporter and mutagenesis of the RUNX1 binding site in the EPHA2 promoter confirmed this regulatory axis. |
Transcriptomic and proteomic analysis, luciferase reporter assay with RUNX1/EPHA2 promoter binding site mutation, VEGF ELISA, tube formation assay, in vivo experiments |
Cancer biology & therapy |
Medium |
39797421
|