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PreprintCited by 12Wnt/β-catenin signaling in development and cancerSignaling Pathways in DiseaseProtein Kinase Regulation and GTPase Signaling

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Protein proximity networks and functional evaluation of the Casein Kinase 1 γ family reveals unique roles for CK1 γ 3 in WNT signaling

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Agajanian, Megan J., Potjewyd, Frances M., Bowman, Brittany M., Solomon, Smaranda, LaPak, Kyle M., Bhatt, Dhaval P., Smith, Jeffrey L., Goldfarb, Dennis, Axtman, Alison D., Major, Michael B.·2021·Journal of Biological Chemistry
DOI 10.1101/2021.11.30.470617PMC PMC9157009

Preprint — published as 10.1016/j.jbc.2022.101986

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Abstract

The WNT/ β -catenin signaling pathway is evolutionarily conserved and controls normal embryonic development, adult tissue homeostasis and regeneration. Aberrant activation or suppression of WNT signaling contributes to cancer initiation and progression, developmental disorders, neurodegeneration, and bone disease. Despite great need and more than 40 years of research, targeted therapies for the WNT pathway have yet to be fully realized. Kinases are exceptionally druggable and occupy key nodes within the WNT signaling network, but several pathway-relevant kinases remain understudied and ‘dark’. Here we studied the function of the CSNK1 γ subfamily of human kinases. miniTurbo-based proximity biotinylation and mass spectrometry analysis of CSNK1 γ 1, CSNK1 γ 2, and CSNK1 γ 3 revealed numerous established components of the β -catenin- dependent and independent WNT signaling pathway, as well as novel interactors. In gain-of- function experiments leveraging a panel of transcriptional reporters, CSNK1 γ 3 but not CSNK1 γ 1 or CSNK1 γ 2 activated β -catenin-dependent WNT signaling and the Notch pathway. Within the family, CSNK1 γ 3 expression uniquely induced LRP6 phosphorylation. Conversely, siRNA- mediated silencing of CSNK1 γ 3 alone had no impact on WNT signaling, though co-silencing of all three family members decreased WNT pathway activity. We characterized two moderately selective and potent small molecule inhibitors of the CSNK1 γ family. These inhibitors and a CSNK1 γ 3 kinase dead mutant suppressed but did not eliminate WNT-driven LRP6 phosphorylation and β -catenin stabilization. Our data suggest that while CSNK1 γ 3 expression uniquely drives pathway activity, potential functional redundancy within the family necessitates loss of all three family members to suppress the WNT signaling pathway.

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Figure 1 CK1γ proximity-based interaction networks identify WNT components.A, schematic of CK1γ family highlighting conserved amino acids and sequence similarity between family members. B, W. blot analysis of HEK293T cells stably expressing mini-Turbo CK1γ or V5 control incubated with 50 mM of biotin over a time course (untreated, 15 min, 30 min, 45 min, 1 h, and 2 h). C, W. blot analysis of streptavidin affinity pulldown from stable HEK293T cells expressing mini-Turbo CK1γ or V5, treated as indicated. D, heat map demonstrating changes in log2(LFQ intensity) of protein proximity partners for the CK1γ subfamily. Red track indicates hits in previously published WNT screens (Tables S1 and S2). Green track indicates well-established proteins involved in WNT/β-catenin-dependent and WNT/β-catenin-independent signaling. E, streptavidin affinity purification mass spectrometry (APMS) proximity networks for CK1γ1, CK1γ2, and CK1γ. HEK293T cells stably expressing mT-V5-CK1γ were treated with 50 mM biotin for 30 min before streptavidin APMS. Blue circle indicates localization to the plasma membrane. Gray lines represent bait–prey interactions. Purple lines indicate prey–prey interactions collected from the BioGRID database. F, W. blot validation of mT-CK1γ protein–protein interaction networks treated with biotin and WNT3A CM for 30 min. CK1γ, casein kinase 1γ; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LFQ, label-free quantitation; W. blot, Western blot.
Figure 1 CK1γ proximity-based interaction networks identify WNT components.A, schematic of CK1γ family highlighting conserved amino acids and sequence similarity between family members. B, W. blot analysis of HEK293T cells stably expressing mini-Turbo CK1γ or V5 control incubated with 50 mM of biotin over a time course (untreated, 15 min, 30 min, 45 min, 1 h, and 2 h). C, W. blot analysis of streptavidin affinity pulldown from stable HEK293T cells expressing mini-Turbo CK1γ or V5, treated as indicated. D, heat map demonstrating changes in log2(LFQ intensity) of protein proximity partners for the CK1γ subfamily. Red track indicates hits in previously published WNT screens (Tables S1 and S2). Green track indicates well-established proteins involved in WNT/β-catenin-dependent and WNT/β-catenin-independent signaling. E, streptavidin affinity purification mass spectrometry (APMS) proximity networks for CK1γ1, CK1γ2, and CK1γ. HEK293T cells stably expressing mT-V5-CK1γ were treated with 50 mM biotin for 30 min before streptavidin APMS. Blue circle indicates localization to the plasma membrane. Gray lines represent bait–prey interactions. Purple lines indicate prey–prey interactions collected from the BioGRID database. F, W. blot validation of mT-CK1γ protein–protein interaction networks treated with biotin and WNT3A CM for 30 min. CK1γ, casein kinase 1γ; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LFQ, label-free quantitation; W. blot, Western blot.CC BY · PubMed Central
Figure 2 Low-throughput reporter screen identifies CK1γ3 as an activator of WNT signaling.A, HEK293T cells were transiently transfected for 30 h with 100 ng of indicated positive control (β-catenin, NRF2, and NICD) construct or pGUS negative control or the indicated CK1γ construct. About 40 ng TK-Renilla and 40 ng of the indicated Response Element–Luciferase construct was included. Where indicated, cells were treated with (TGFβ 10 ng/ml, TNFα 10 ng/ml, and retinoic acid [RA] 10 ng/ml) for 6 h (TGFβ and TNFα) or 16 h (RA). Inset W. blot demonstrates protein overexpression for the BAR luciferase assay. Each condition was normalized to pGUS control. Error bars represent standard deviation, and statistics are compared to pGUS control. B, BAR luciferase assay from HEK293T B/R (stable BAR-firefly luciferase, TK-Renilla–expressing cell line) cells transfected for 24 h with indicated DNA mass. Statistics are compared to pGUS control. Inset blot is confirmation of overexpression for luciferase assay in B. Dashed line represents cropped blot for removal of redundant untreated sample. C, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with Lcell or WNT3A CM for 16 h. All statistics are compared to FLAG-Control Lcell sample. D, HEK293T B/R cells were transfected with the indicated construct for 24 h before luciferase quantitation. Statistics are compared to pGUS control. E, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with 10 μM C59 for 18 h. F, WT, LRP5/6 DKO, DVL TKO HEK293T cells transfected with 20 ng BAR-luciferase, 10 ng TK-Renilla, and 70 ng of indicated constructs for 14 h, and then treated with either Lcell CM or WNT3A CM for 18 h. Statistical significance presented is compared with Lcell-treated FLAG-Control cells for each cell type. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All data are plotted as box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ3, casein kinase 1γ3; CM, conditioned media; DKO, double KO; DVL, disheveled; HEK293T, human embryonic kidney 293T cell line; LRP, low-density lipoprotein receptor–related protein; NICD, Notch intracellular domain; NRF2, nuclear factor erythroid 2–related factor 2; TGFβ, transforming growth factor beta; TKO, triple KO; TNFα, tumor necrosis factor alpha; W. blot, Western blot.
Figure 2 Low-throughput reporter screen identifies CK1γ3 as an activator of WNT signaling.A, HEK293T cells were transiently transfected for 30 h with 100 ng of indicated positive control (β-catenin, NRF2, and NICD) construct or pGUS negative control or the indicated CK1γ construct. About 40 ng TK-Renilla and 40 ng of the indicated Response Element–Luciferase construct was included. Where indicated, cells were treated with (TGFβ 10 ng/ml, TNFα 10 ng/ml, and retinoic acid [RA] 10 ng/ml) for 6 h (TGFβ and TNFα) or 16 h (RA). Inset W. blot demonstrates protein overexpression for the BAR luciferase assay. Each condition was normalized to pGUS control. Error bars represent standard deviation, and statistics are compared to pGUS control. B, BAR luciferase assay from HEK293T B/R (stable BAR-firefly luciferase, TK-Renilla–expressing cell line) cells transfected for 24 h with indicated DNA mass. Statistics are compared to pGUS control. Inset blot is confirmation of overexpression for luciferase assay in B. Dashed line represents cropped blot for removal of redundant untreated sample. C, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with Lcell or WNT3A CM for 16 h. All statistics are compared to FLAG-Control Lcell sample. D, HEK293T B/R cells were transfected with the indicated construct for 24 h before luciferase quantitation. Statistics are compared to pGUS control. E, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with 10 μM C59 for 18 h. F, WT, LRP5/6 DKO, DVL TKO HEK293T cells transfected with 20 ng BAR-luciferase, 10 ng TK-Renilla, and 70 ng of indicated constructs for 14 h, and then treated with either Lcell CM or WNT3A CM for 18 h. Statistical significance presented is compared with Lcell-treated FLAG-Control cells for each cell type. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All data are plotted as box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ3, casein kinase 1γ3; CM, conditioned media; DKO, double KO; DVL, disheveled; HEK293T, human embryonic kidney 293T cell line; LRP, low-density lipoprotein receptor–related protein; NICD, Notch intracellular domain; NRF2, nuclear factor erythroid 2–related factor 2; TGFβ, transforming growth factor beta; TKO, triple KO; TNFα, tumor necrosis factor alpha; W. blot, Western blot.CC BY · PubMed Central
Figure 3 Overexpression of CK1γ3 activates WNT signaling by increasing LRP6 phosphorylation.A, HEK293T cells were transfected with the indicated construct for 14 h and then exposed to WNT3A CM for indicated time and analyzed by W. blot. B and C, quantification of W. blots from A at indicated time points. pLRP6 T1479 (B) and pLRP6 S1490 (C) across four replicates, normalized to total LRP6. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All data are plotted as box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. All statistics are compared to FLAG-Control untreated sample. CK1γ3, casein kinase 1γ3; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LRP6, low-density lipoprotein receptor–related protein 6; W. blot, Western blot.
Figure 3 Overexpression of CK1γ3 activates WNT signaling by increasing LRP6 phosphorylation.A, HEK293T cells were transfected with the indicated construct for 14 h and then exposed to WNT3A CM for indicated time and analyzed by W. blot. B and C, quantification of W. blots from A at indicated time points. pLRP6 T1479 (B) and pLRP6 S1490 (C) across four replicates, normalized to total LRP6. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All data are plotted as box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. All statistics are compared to FLAG-Control untreated sample. CK1γ3, casein kinase 1γ3; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LRP6, low-density lipoprotein receptor–related protein 6; W. blot, Western blot.CC BY · PubMed Central
Figure 4 CK1γ3 kinase activity is required for maximum WNT3A activation.A, HEK293T B/R cells were transfected with the indicated construct for 14 h, treated with Lcell or WNT3A CM for 18 h, and then analyzed by luciferase assay. B, live-cell imaging of HEK293T cells stably expressing a BAR-GFP fluorescent reporter transiently transfected with the indicated expression construct, CK1γ3-WT or CK1γ3-K72A. Lcell or WNT3A CM was added at 8 h, and cells were monitored for an additional 72 h. Data are averaged across four replicates. C, HEK293T cells transfected with either CK1γ3-WT or CK1γ3-K72A for 14 h then either untreated or treated with WNT3A CM for 30 or 60 min. Samples were then analyzed by W. blot. D and E, quantification of W. blot from A for indicated time points, pLRP6 T1479 (D) and pLRP6 S1490 (E) across four biological replicates, normalized to total LRP6. All statistics are compared to FLAG-CK1γ3 Lcell sample. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ3, casein kinase 1γ3; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LRP6, low-density lipoprotein receptor–related protein 6; W, blot, Western blot.
Figure 4 CK1γ3 kinase activity is required for maximum WNT3A activation.A, HEK293T B/R cells were transfected with the indicated construct for 14 h, treated with Lcell or WNT3A CM for 18 h, and then analyzed by luciferase assay. B, live-cell imaging of HEK293T cells stably expressing a BAR-GFP fluorescent reporter transiently transfected with the indicated expression construct, CK1γ3-WT or CK1γ3-K72A. Lcell or WNT3A CM was added at 8 h, and cells were monitored for an additional 72 h. Data are averaged across four replicates. C, HEK293T cells transfected with either CK1γ3-WT or CK1γ3-K72A for 14 h then either untreated or treated with WNT3A CM for 30 or 60 min. Samples were then analyzed by W. blot. D and E, quantification of W. blot from A for indicated time points, pLRP6 T1479 (D) and pLRP6 S1490 (E) across four biological replicates, normalized to total LRP6. All statistics are compared to FLAG-CK1γ3 Lcell sample. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ3, casein kinase 1γ3; CM, conditioned media; HEK293T, human embryonic kidney 293T cell line; LRP6, low-density lipoprotein receptor–related protein 6; W, blot, Western blot.CC BY · PubMed Central
Figure 5 Loss of CK1γ1/2/3 impairs WNT activation.A–C, RKO B/R cells were transfected with indicated siRNA or pooled siRNA for 48 h and then split for the following experiments: 24 h postsplit gene expression was analyzed by RT–qPCR (A), 6 h postsplit cells were treated with WNT3A for 18 h and then analyzed by luciferase assay (B), 24 h postsplit cells were exposed to a WNT3A time course, harvested at the indicated time post WNT3A, and analyzed by W. blot. For A, all statistics are compared to siControl expression for the specific gene. For B, all statistics are compared to siControl WNT3A-treated cells. D–F, quantification of W. blot from E, β-catenin (D), pLRP6 T1479 (E), and pLRP6 S1490 (F) across four replicates, normalized to actin. Statistics are compared to siControl untreated cells, unless otherwise stated. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ, casein kinase 1γ; qPCR, quantitative PCR; W. blot, Western blot.
Figure 5 Loss of CK1γ1/2/3 impairs WNT activation.A–C, RKO B/R cells were transfected with indicated siRNA or pooled siRNA for 48 h and then split for the following experiments: 24 h postsplit gene expression was analyzed by RT–qPCR (A), 6 h postsplit cells were treated with WNT3A for 18 h and then analyzed by luciferase assay (B), 24 h postsplit cells were exposed to a WNT3A time course, harvested at the indicated time post WNT3A, and analyzed by W. blot. For A, all statistics are compared to siControl expression for the specific gene. For B, all statistics are compared to siControl WNT3A-treated cells. D–F, quantification of W. blot from E, β-catenin (D), pLRP6 T1479 (E), and pLRP6 S1490 (F) across four replicates, normalized to actin. Statistics are compared to siControl untreated cells, unless otherwise stated. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. CK1γ, casein kinase 1γ; qPCR, quantitative PCR; W. blot, Western blot.CC BY · PubMed Central
Figure 6 Summary of data for AKI00000062a.A, structure of AKI00000062a. B, enzymatic profiling of AKI00000062a versus CK1γ family kinases. C, kinome tree representation of kinases with PoC <10 when AKI00000062a was profiled at 1 μM versus 403 WT kinases. D, specific kinases (n = 17) with PoC <10 in kinome-wide profiling. E, Nanoluc-CK1γ2 NanoBRET curve, and corresponding IC50 value for AKI00000062a. CK1γ, casein kinase 1γ; PoC, Percent of Control.
Figure 6 Summary of data for AKI00000062a.A, structure of AKI00000062a. B, enzymatic profiling of AKI00000062a versus CK1γ family kinases. C, kinome tree representation of kinases with PoC <10 when AKI00000062a was profiled at 1 μM versus 403 WT kinases. D, specific kinases (n = 17) with PoC <10 in kinome-wide profiling. E, Nanoluc-CK1γ2 NanoBRET curve, and corresponding IC50 value for AKI00000062a. CK1γ, casein kinase 1γ; PoC, Percent of Control.CC BY · PubMed Central
Figure 7 Summary of data for FP1-24-2.A, structure of FP1-24-2. B, enzymatic profiling of FP1-24-2 against CK1γ family kinases. C, kinome tree representation of kinases with PoC <10 when FP1-24-2 was profiled at 1 μM versus 403 WT kinases. D, specific kinases (n = 14) with PoC <10 in kinome-wide profiling. E, Nanoluc-CK1γ2 NanoBRET curve and corresponding IC50 value for FP1-24-2. CK1γ, casein kinase 1γ; PoC, Percent of Control.
Figure 7 Summary of data for FP1-24-2.A, structure of FP1-24-2. B, enzymatic profiling of FP1-24-2 against CK1γ family kinases. C, kinome tree representation of kinases with PoC <10 when FP1-24-2 was profiled at 1 μM versus 403 WT kinases. D, specific kinases (n = 14) with PoC <10 in kinome-wide profiling. E, Nanoluc-CK1γ2 NanoBRET curve and corresponding IC50 value for FP1-24-2. CK1γ, casein kinase 1γ; PoC, Percent of Control.CC BY · PubMed Central
Figure 8 CK1γ inhibition suppresses β-catenin stabilization and LRP6 phosphorylation.A and B, Nanoluc-GSK3β NanoBRET curve and corresponding IC50 value for AKI and FP1, respectively. C, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with 10 μM of the indicated compound for 18 h and analyzed by luciferase assay. D, HEK293T B/R cells were transfected with the indicated construct for 14 h, treated with DMSO, 0.1, 1.0, or 10 μM of FP1 for 18 h, and then analyzed by luciferase assay. E, RKO cells were pretreated with FP1 or DMSO for 1 h, treated with WNT3A ligand for 30 or 60 min, and then analyzed by W. blot. Dashed line represents cropped blot for removal of redundant untreated sample. F–H, quantification of W. blot from E, β-catenin (F), pLRP6 S1490 (G), and pLRP6 T1479 (H) across four replicates, normalized to actin. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05 and are compared to DMSO control unless otherwise indicated. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. AKI, AKI00000062a; CK1γ, casein kinase 1γ; DMSO, dimethyl sulfoxide; FP1, FP1-24-2; GSK3β, glycogen synthase kinase 3β; HEK293T, human embryonic kidney 293T cell line; W. blot, Western blot.
Figure 8 CK1γ inhibition suppresses β-catenin stabilization and LRP6 phosphorylation.A and B, Nanoluc-GSK3β NanoBRET curve and corresponding IC50 value for AKI and FP1, respectively. C, HEK293T B/R cells were transfected with the indicated construct for 14 h and then treated with 10 μM of the indicated compound for 18 h and analyzed by luciferase assay. D, HEK293T B/R cells were transfected with the indicated construct for 14 h, treated with DMSO, 0.1, 1.0, or 10 μM of FP1 for 18 h, and then analyzed by luciferase assay. E, RKO cells were pretreated with FP1 or DMSO for 1 h, treated with WNT3A ligand for 30 or 60 min, and then analyzed by W. blot. Dashed line represents cropped blot for removal of redundant untreated sample. F–H, quantification of W. blot from E, β-catenin (F), pLRP6 S1490 (G), and pLRP6 T1479 (H) across four replicates, normalized to actin. For all panels: ∗∗∗p < 0.0005, ∗∗p < 0.005, and ∗p < 0.05 and are compared to DMSO control unless otherwise indicated. All biological replicates are represented in box-and-whisker plots, with the boxes representing the median and the interquartile range. All biological replicates are averaged across three technical replicates. Statistical significance was evaluated by Student’s t test. AKI, AKI00000062a; CK1γ, casein kinase 1γ; DMSO, dimethyl sulfoxide; FP1, FP1-24-2; GSK3β, glycogen synthase kinase 3β; HEK293T, human embryonic kidney 293T cell line; W. blot, Western blot.CC BY · PubMed Central

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Metadata

Authors
Agajanian, M. J., Potjewyd, F. M., Bowman, B. M., Solomon, S., LaPak, K. M., Bhatt, D. P., Smith, J. L., Goldfarb, D., Axtman, A. D., & Major, M. B.
Affiliations
  • University of North Carolina at Chapel Hill (US)
  • Washington University in St. Louis (US)
Corresponding author
Major, Michael B.
ORCID
Venue
Journal of Biological Chemistry
Publisher
openRxiv
Published
2021-11-30
Volume
298
Issue
6
Pages
101986-101986
ISSN
0021-9258 (ISSN-L)
DOI
10.1101/2021.11.30.470617
PMC
PMC9157009
OpenAlex
W4224939721
Cited by
12
References
1
Citations by year
2026: 6 · 2025: 1 · 2024: 1 · 2023: 4
FWCI
0.49
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Commons@Becker (Washington University School of Medicine) (submittedVersion, cc-by)
Funders
  • Howard Hughes Medical Institute
  • Boehringer Ingelheim
  • Pfizer
  • North Carolina Biotechnology Center (2018-IDG-1030)
  • Meso Scale Diagnostics
  • Takeda Pharmaceuticals U.S.A.
  • Genome Canada (115766)
  • Merck KGaA
  • Wellcome Trust (106169/ZZ14/Z)
  • European Federation of Pharmaceutical Industries and Associations (115766)
  • Ontario Ministry of Economic Development and Innovation
  • Canada Foundation for Innovation
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (106169/ZZ14/Z)
  • Ministero dello Sviluppo Economico
  • Innovative Medicines Initiative
  • National Institutes of Health (5f99ca245724-02, 1097737, F99/K00)
  • Genentech
  • Novartis Pharma
  • National Institute on Aging (U54AG065187)
  • National Cancer Institute (F99 CA245724, K00CA245724, F99/K00, T32CA009547)
  • National Institute of Diabetes and Digestive and Kidney Diseases (U24DK116204)
  • Janssen Pharmaceuticals
  • Eshelman Institute for Innovation, University of North Carolina at Chapel Hill
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Signaling Pathways in Disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
Wnt/β-catenin signaling, protein-protein interaction network, LRP6 phosphorylation, β-catenin stabilization, kinase inhibitors, functional redundancy, kinase families, cancer signaling
MeSH terms
Humans, Phosphorylation, Casein Kinase I, Wnt Proteins, beta Catenin, Wnt Signaling Pathway, Low Density Lipoprotein Receptor-Related Protein-6

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CoChat. (2026). Citation check: Protein proximity networks and functional evaluation of the Casein Kinase 1 γ family reveals unique roles for CK1 γ 3 in WNT signaling. https://app.cochat.ai/papers/10.1101/2021.11.30.470617
The paper
Agajanian, M. J., Potjewyd, F. M., Bowman, B. M., Solomon, S., LaPak, K. M., Bhatt, D. P., Smith, J. L., Goldfarb, D., Axtman, A. D., & Major, M. B. (2021). Protein proximity networks and functional evaluation of the Casein Kinase 1 γ family reveals unique roles for CK1 γ 3 in WNT signaling. Journal of Biological Chemistry. https://doi.org/10.1101/2021.11.30.470617