Affinage

CSNK1G3

Casein kinase I isoform gamma-3 · UniProt Q9Y6M4

Round 2 corrected
Length
447 aa
Mass
51.4 kDa
Annotated
2026-04-28
42 papers in source corpus 7 papers cited in narrative 7 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CSNK1G3 (CK1γ3) is a serine/threonine kinase of the casein kinase 1 gamma subfamily that autophosphorylates and phosphorylates canonical CK1 substrates (casein, phosvitin), and functionally complements yeast CK1 orthologs YCK1/YCK2 (PMID:7759525). CSNK1G3 activates Wnt/β-catenin signaling through a Dishevelled-independent mechanism distinct from other CK1 isoforms (PMID:11437445) and supports AKT Ser-473 phosphorylation and downstream mTOR/S6 signaling, linking it to epithelial–mesenchymal transition and survival in cancer cells (PMID:16247451, PMID:39127154). Its expression is positively regulated at the post-transcriptional level by circRNA-Csnk1g3 targeting the 3′-UTR of CSNK1G3 mRNA, and elevated CSNK1G3 promotes Wnt/β-catenin-driven pyroptosis and RIP1/RIP3/MLKL-mediated necroptosis in hippocampal neurons (PMID:41633109, PMID:39920301).

Mechanistic history

Synthesis pass · year-by-year structured walk · 6 steps
  1. 1995 High

    Cloning and biochemical characterization established CSNK1G3 as an enzymatically active CK1 family kinase with broad tissue expression, resolving the basic identity and catalytic competence of the gamma-3 isoform.

    Evidence Recombinant protein expressed in E. coli phosphorylated canonical CK1 substrates and autophosphorylated; yeast complementation of YCK1/YCK2 deletion confirmed in vivo kinase function

    PMID:7759525

    Open questions at the time
    • No endogenous substrate identified in mammalian cells
    • Subcellular localization not determined
    • Regulation of kinase activity (beyond CK1-7 inhibition) not addressed
  2. 2001 Medium

    Demonstration that CK1γ isoforms, including CSNK1G3, activate Wnt/β-catenin signaling without phosphorylating Dishevelled revealed a mechanistically distinct entry point for the gamma subfamily into Wnt pathway regulation.

    Evidence Wnt reporter assays showed activation by CK1γ3; in vitro kinase assays and in vivo phosphorylation studies showed no Dsh phosphorylation by gamma isoforms

    PMID:11437445

    Open questions at the time
    • The direct substrate mediating Wnt activation by CK1γ3 was not identified
    • Physiological relevance not confirmed by loss-of-function in vivo
    • Relationship to LRP6 phosphorylation not yet tested
  3. 2006 Medium

    A kinome-wide RNAi screen placed CSNK1G3 upstream of AKT Ser-473 and ribosomal protein S6 phosphorylation, establishing it as a cooperating kinase in AKT-dependent survival signaling in cancer cells.

    Evidence siRNA knockdown reduced p-AKT(S473) and p-S6 levels and sensitized cancer cells to an AKT inhibitor

    PMID:16247451

    Open questions at the time
    • Whether CSNK1G3 directly phosphorylates AKT or acts indirectly was not resolved
    • Only one cell line context; generalizability unknown
    • No reconstituted kinase-substrate assay for the AKT axis
  4. 2024 Medium

    Identification of CSNK1G3 as the direct target of the curcumin derivative N17 independently confirmed the CSNK1G3–AKT signaling axis and extended it to EMT regulation and apoptosis in triple-negative breast cancer.

    Evidence Target engagement of N17 with CSNK1G3 validated by binding assays; inhibition suppressed p-AKT(S473), EMT markers, and triggered apoptosis

    PMID:39127154

    Open questions at the time
    • Direct phosphorylation of AKT by CSNK1G3 still not biochemically reconstituted
    • Selectivity of N17 for CSNK1G3 over other CK1 isoforms not fully characterized
    • In vivo tumor model validation not reported
  5. 2025 Low

    Discovery that circRNA-Csnk1g3 upregulates CSNK1G3 protein via 3′-UTR targeting, and that elevated CK1γ3 drives TNF-α/RIP1/RIP3/MLKL necroptosis in hippocampal neurons, placed CSNK1G3 in neuroinflammatory cell death for the first time.

    Evidence Mouse epilepsy model and HT22 cells; qRT-PCR, western blot, TUNEL, flow cytometry for necroptosis and inflammatory markers

    PMID:39920301

    Open questions at the time
    • CK1γ3 substrates in the necroptosis pathway not identified; pathway placement is correlative
    • Single lab, no independent replication
    • Relative contribution of truncated Csnk1g3-85aa isoform versus full-length kinase unclear
  6. 2026 Medium

    Orthogonal validation by dual-luciferase and RIP assays confirmed the circRNA-Csnk1g3/CSNK1G3 regulatory axis and extended the downstream consequence to Wnt/β-catenin-mediated pyroptosis (NLRP3/Caspase-1/GSDMD) in epileptic neurons.

    Evidence Dual-luciferase reporter and RNA immunoprecipitation in HT22 cells; pyroptosis markers by western blot, immunofluorescence, and electron microscopy

    PMID:41633109

    Open questions at the time
    • Whether CK1γ3 directly phosphorylates LRP6 or β-catenin pathway components in this context is unknown
    • Only one neuronal cell line used; in vivo pyroptosis phenotype not confirmed with CSNK1G3 genetic ablation
    • Relationship between necroptosis (TNF-α axis) and pyroptosis (Wnt axis) downstream of CK1γ3 not delineated

Open questions

Synthesis pass · forward-looking unresolved questions
  • The direct phosphorylation substrates by which CSNK1G3 activates Wnt/β-catenin signaling and supports AKT Ser-473 phosphorylation remain unidentified, and the structural basis for isoform-specific functions within the CK1γ subfamily is unresolved.
  • No crystal structure or cryo-EM model of CSNK1G3
  • No unbiased phosphoproteomics identifying direct substrates
  • In vivo loss-of-function phenotype in knockout mice not reported

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0140096 catalytic activity, acting on a protein 4 GO:0016740 transferase activity 1
Pathway
R-HSA-162582 Signal Transduction 4 R-HSA-5357801 Programmed Cell Death 3
Partners
AKT1CTNNB1MLKLRIP1RIP3

Evidence

Reading pass · 7 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1995 CK1γ3 (CSNK1G3) was molecularly cloned from rat testis as one of three members of the CK1 gamma subfamily. The recombinant protein expressed in E. coli was shown to be an active kinase that phosphorylates canonical CK1 substrates (casein, phosvitin, and the synthetic peptide D4), undergoes autophosphorylation in the presence of ATP and Mg2+, and is inhibited by the CK1 inhibitor CK1-7. CK1γ3 mRNA was detected in brain, heart, kidney, lung, liver, and muscle. Expression of CK1γ1 or CK1γ3 complemented defects in yeast YCK1/YCK2 mutants, restoring growth and morphology. Molecular cloning, recombinant protein expression in E. coli, in vitro kinase assay, Northern hybridization, yeast complementation assay The Journal of biological chemistry High 7759525
2001 All wild-type CK1 isoforms, including the gamma subfamily members (CK1γ1, γ2, γ3), were shown to activate Wnt/β-catenin signaling in sufficiency tests. Unlike other CK1 isoforms, the gamma isoforms did not increase phosphorylation of Dishevelled (Dsh) in vivo and did not directly phosphorylate Dsh in vitro, suggesting a distinct mechanism by which CK1γ isoforms contribute to Wnt signaling. In vivo Wnt signaling reporter assay, in vitro kinase assay with Dsh substrate, cell-based phosphorylation assay Developmental biology Medium 11437445
2006 An siRNA kinome screen identified CSNK1G3 as an Akt-cooperating kinase in cancer cells. siRNA knockdown of CSNK1G3 enhanced cell killing by the Akt inhibitor A-443654 and caused decreases in Akt Ser-473 and ribosomal protein S6 phosphorylation, placing CSNK1G3 in a signaling axis that supports Akt activity and downstream mTOR/S6 signaling. RNAi kinome screen (siRNA library), cell viability assay, immunoblot for phospho-Akt(S473) and phospho-S6 Oncogene Medium 16247451
2018 siRNA knockdown of CSNK1G3 was found to negatively regulate translation readthrough at premature stop codons in a dual reporter-based high-throughput screen, indicating that CSNK1G3 activity promotes readthrough and competes with translation termination. High-throughput siRNA screen using dual reporter (enzymatic and fluorescence activities), functional readthrough assay Human molecular genetics Low 29177465
2024 CSNK1G3 was identified as the direct molecular target of the curcumin derivative N17 in triple-negative breast cancer cells. CSNK1G3 was shown to regulate phosphorylation of AKT at Ser-473 (p-AKT(S473)), and its inhibition by N17 suppressed the epithelial-mesenchymal transition (EMT) and triggered apoptosis through the CSNK1G3/AKT signaling axis. Target identification (N17 compound binding to CSNK1G3), immunoblot for p-AKT(S473), cell proliferation and apoptosis assays, EMT marker analysis Biochemical pharmacology Medium 39127154
2025 In a mouse epilepsy model and HT22 hippocampal neuronal cells, circRNA-Csnk1g3 was shown to promote transcription of the canonical Csnk1g3 mRNA (and a truncated isoform Csnk1g3-85aa) by directly targeting its 3'-UTR. Elevated Csnk1g3/CK1γ3 protein was associated with increased TNF-α signaling, RIP1/RIP3/MLKL-mediated necroptosis, and neuroinflammation (IL-6, IL-1β). The traditional medicine JWCHSGD reduced epileptic events by downregulating this circRNA-Csnk1g3/Csnk1g3-85aa/CK1γ3/TNF-α axis. In vivo epilepsy mouse model, HT22 cell culture, EEG, TUNEL assay, qRT-PCR, western blot, CCK-8 viability, flow cytometry apoptosis assay Scientific reports Low 39920301
2026 In hippocampal HT22 cells treated with a magnesium-free epilepsy model, circRNA-Csnk1g3 was shown by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) to directly target the 3'-UTR of Csnk1g3 mRNA and promote its transcription. Elevated Csnk1g3 drove pyroptosis via the Wnt/β-catenin pathway (upregulating Wnt3a, β-catenin, LRP6, Caspase-1, GSDMD, NLRP3, IL-1β). Knockdown of circRNA-Csnk1g3 reduced Csnk1g3 expression and pyroptotic markers. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), FISH for subcellular localization, qRT-PCR, western blot, immunofluorescence, electron microscopy, Dil staining, LC-MS/MS chemical characterization Journal of ethnopharmacology Medium 41633109

Source papers

Stage 0 corpus · 42 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2013 Discovery and refinement of loci associated with lipid levels. Nature genetics 2409 24097068
2002 Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proceedings of the National Academy of Sciences of the United States of America 1479 12477932
2010 Network organization of the human autophagy system. Nature 1286 20562859
2015 The BioPlex Network: A Systematic Exploration of the Human Interactome. Cell 1118 26186194
2017 Architecture of the human interactome defines protein communities and disease networks. Nature 1085 28514442
2015 A human interactome in three quantitative dimensions organized by stoichiometries and abundances. Cell 1015 26496610
2020 A reference map of the human binary protein interactome. Nature 849 32296183
2021 Dual proteome-scale networks reveal cell-specific remodeling of the human interactome. Cell 705 33961781
2011 Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium. Briefings in bioinformatics 656 21873635
2004 The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC). Genome research 438 15489334
2022 OpenCell: Endogenous tagging for the cartography of human cellular organization. Science (New York, N.Y.) 432 35271311
2005 Diversification of transcriptional modulation: large-scale identification and characterization of putative alternative promoters of human genes. Genome research 409 16344560
2021 A proximity-dependent biotinylation map of a human cell. Nature 339 34079125
2004 Transcriptome characterization elucidates signaling networks that control human ES cell growth and differentiation. Nature biotechnology 266 15146197
2009 Reduced expression of the Kinesin-Associated Protein 3 (KIFAP3) gene increases survival in sporadic amyotrophic lateral sclerosis. Proceedings of the National Academy of Sciences of the United States of America 149 19451621
2009 Ubiquitin-mediated proteolysis of HuR by heat shock. The EMBO journal 142 19322201
2019 Mapping the proximity interaction network of the Rho-family GTPases reveals signalling pathways and regulatory mechanisms. Nature cell biology 137 31871319
2017 RNA-binding activity of TRIM25 is mediated by its PRY/SPRY domain and is required for ubiquitination. BMC biology 135 29117863
2001 The casein kinase I family in Wnt signaling. Developmental biology 104 11437445
2005 The role of the casein kinase 1 (CK1) family in different signaling pathways linked to cancer development. Onkologie 102 16186692
2022 EZH2 depletion potentiates MYC degradation inhibiting neuroblastoma and small cell carcinoma tumor formation. Nature communications 99 35013218
1995 Casein kinase I gamma subfamily. Molecular cloning, expression, and characterization of three mammalian isoforms and complementation of defects in the Saccharomyces cerevisiae YCK genes. The Journal of biological chemistry 99 7759525
2020 Kinase Interaction Network Expands Functional and Disease Roles of Human Kinases. Molecular cell 88 32707033
2014 Phosphorylation of LRRK2 by casein kinase 1α regulates trans-Golgi clustering via differential interaction with ARHGEF7. Nature communications 86 25500533
2015 Systematic proteomics of the VCP-UBXD adaptor network identifies a role for UBXN10 in regulating ciliogenesis. Nature cell biology 81 26389662
2019 The midbody interactome reveals unexpected roles for PP1 phosphatases in cytokinesis. Nature communications 74 31586073
2018 Interrogating the protein interactomes of RAS isoforms identifies PIP5K1A as a KRAS-specific vulnerability. Nature communications 73 30194290
2021 Histone deacetylase inhibitors inhibit cervical cancer growth through Parkin acetylation-mediated mitophagy. Acta pharmaceutica Sinica. B 66 35256949
2007 Arabidopsis HY5 protein functions as a DNA-binding tag for purification and functional immobilization of proteins on agarose/DNA microplate. FEBS letters 65 18082144
2016 Estimation of genetic parameters and detection of chromosomal regions affecting the major milk proteins and their post translational modifications in Danish Holstein and Danish Jersey cattle. BMC genetics 53 27485317
2020 RIG-I regulates myeloid differentiation by promoting TRIM25-mediated ISGylation. Proceedings of the National Academy of Sciences of the United States of America 47 32513696
2006 RNAi-based screening of the human kinome identifies Akt-cooperating kinases: a new approach to designing efficacious multitargeted kinase inhibitors. Oncogene 44 16247451
2018 Aminoglycoside-mediated promotion of translation readthrough occurs through a non-stochastic mechanism that competes with translation termination. Human molecular genetics 18 29177465
2021 Detecting Genetic Ancestry and Adaptation in the Taiwanese Han People. Molecular biology and evolution 14 33170928
2020 Autoimmune response against tyrosinase induces depigmentation in C57BL/6 black mice. Autoimmunity 13 33084421
2017 Gene-centric analysis implicates nuclear encoded mitochondrial protein gene variants in migraine susceptibility. Molecular genetics & genomic medicine 6 28361102
2025 Differential analysis of testicular LncRNA in Kazakh horses of different ages. International journal of biological macromolecules 3 40706934
2024 Novel curcumin derivatives N17 exert anti-cancer effects through the CSNK1G3/AKT axis in triple-negative breast cancer. Biochemical pharmacology 2 39127154
2025 Mitigating effects of Jiawei Chaihu Shugan decoction on necroptosis and inflammation of hippocampal neurons in epileptic mice. Scientific reports 1 39920301
2025 Clinical Outcomes of Patients With Advanced ALK-Rearranged Lung Squamous Cell Carcinoma Treated With ALK Tyrosine Kinase Inhibitors. Clinical lung cancer 1 40783309
2025 Dietary Influence on Urolithiasis Risk Mediated by Plasma Metabolites: A Mendelian Randomization and Experimental Study Linking Genes, Metabolites, and Clinical Outcomes. Food science & nutrition 1 40842671
2026 Investigating the effect of Dingxian Pill on hippocampal neuronal pyroptosis in epilepsy through regulation of the circRNA-Csnk1g3/Wnt/β-catenin pathway. Journal of ethnopharmacology 0 41633109