(A) Pathway illustrations of different PKA activating genomic alterations in order from top: PRKACA amplification, DNAJB1-PRKACA fusion, PRKACA activating mutation, and PRKAR1A inactivation or …
Images for Figure 1A.
Table for Figure 1B.
Images for Figure 1D part 1/3.
Images for Figure 1D part 2/3.
Images for Figure 1D part 3/3.
(A) Global phosphorylation changes in 639V with induction of 3xFLAG-PRKACA or 3xFLAG-PRKAR1AG325D; VASP is shown as a positive control for PKA activation, technical replicates shown. (B) Change in …
Tables for Figure 2A.
Tables for Figure 2B.
Tables for Figure 2C.
Tables for Figure 2D.
Tables for Figure 2E.
Images for Figure 2F.
(A) Immunoblots showing the change of PKA activity, as indicated by phospho-PKA substrate, and c-MYC and n-MYC expression in Colo741 and FLX1 cells after treatment with 50 μM forskolin (FSK) and …
Images for Figure 3A.
Tables for Figure 3B.
Images for Figure 3C.
Images for Figure 3D.
Images for Figure 3E.
Images for Figure 3F.
Tables for Figure 3G.
(A) Impact of individual PRKACA-targeting siRNAs on relative cell confluence in FLX1. Confirmation of knockdown shown on right. Experiment was done in duplicate, the representative results shown …
Tables for Figure 3—figure supplement 1A.
Tables for Figure 3—figure supplement 1B.
(A) RNASEQ data from FLX1 cells after 48 hr treatment with four pooled siRNA against PRKACA. PRKACA, ornithine decarboxylase (ODC) and Cyclin D1 (CCND1) are highlighted. (B) Gene set enrichment …
Tables for Figure 4A.
Tables for Figure 4B.
Tables for Figure 4C.
Tables for Figure 4D.
Tables for Figure 4E.
Images for Figure 4E.
Tables for Figure 4F.
Tables for Figure 4G.
Images for Figure 4G.
Tables for Figure 4—figure supplement 1A.
Tables for Figure 4—figure supplement 1B.
Images for Figure 4—figure supplement 1B.
(A) Summary data from the FLX1 cell line treated with 352 kinase inhibitors from an advanced clinical compound library at 2 μM for 120 hr. The targets of selected compounds with a z-score ≥2 are …
Tables for Figure 5A.
Tables for Figure 5B.
Tables for Figure 5C.
Tables for Figure 5D.
Tables for Figure 5E.
Images for Figure 5F.
(A) Colo741 cells treated with dose curves of multiple AURKA inhibitors for 72 hr. Relative cell viability was measured by CTG assay vs. untreated control samples. Results are the mean ± SEM of …
Tables for Figure 5—figure supplement 1A.
Images for Figure 5—figure supplement 1B.
Images for Figure 5—figure supplement 1C.
(B) Immunoblots showing the change of c-MYC level in engineered FLX1 cells with doxycycline (dox)-inducible 3xFLAG- PRKAR1AG325D after dox for 24 hr and/or 20 μM MG132 for 2 hr. (C) Li-Cor western …
Images for Figure 5—figure supplement 2A.
Images for Figure 5—figure supplement 2B.
Images and tables for Figure 5—figure supplement 2C.
(A) Gene set enrichment analysis (GSEA) of significantly altered phosphoproteins following doxycycline (dox) induction of PKA in proteomics data from this study or chemical PKA stimulation in our …
Tables for Figure 6A.
Images for Figure 6B.
Images for Figure 6C.
Images for Figure 6D part 1/4.
Images for Figure 6D part 2/4.
Images for Figure 6D part 3/4.
Images for Figure 6D part 4/4.
(A) Immunoblots showing the c-MYC protein levels in FLX1 cells after treatment with 100 nM rocaglamide or zotatifin for 24 hr and/or 50 μM forskolin (FSK)/3-isobutyl-1-methylxanthine (IBMX) for 4 …
Images for Figure 7A.
Images for Figure 7B.
Images for Figure 7C.
Tables for Figure 7D.
Tables for Figure 7E.
Tables for Figure 7F.
Tables for Figure 7G.
Images for Figure 7H.
(A) Immunoblot showing the effect of 48 hr of non-targeting control (NTC) or four pooled PRKACA siRNA on endogenous c-MYC or overexpressed c-MYC lacking a 5’UTR. Doxycycline (dox) treatment was 1 …
Images for Figure 7—figure supplement 1A.
Images for Figure 7—figure supplement 1B.
Tables for Figure 7—figure supplement 1A.
(A) FLX1 parental cells were treated with DMSO or 50 μM FSK/IBMX for 4 hours, with BrdU added for the last 20 minutes. Cells were then stained for nuclear content with 7-AAD and active DNA synthesis …
Dox treatment was 1 μG/mL for 40 hours prior to harvest with zotatifin or DMSO for 24 hours. Of note, the MYCT58A allele was toxic in both cells but particularly in FLX1, resulting in reduced …
CRISPR | ||
---|---|---|
PRKACA | MYC | |
Average | -0.24 | -1.46 |
639V | -0.14 | -0.67 |
Colo741 | -0.35 | -1.38 |
siRNA | ||
PRKACA | MYC | |
Average | -0.15 | -0.48 |
Colo741 | -1.38 | -0.35 |
Summary of TCGA analysis.
Individual phosphoproteomic, phosfate, and multiplex inhibitor beads (MIBs) datasets.
Network propagation results.
RNASEQ primary data.
Drug screen in FLX1.
Drug screen in FLX1 with dox-inducible PRKAR1AG325D.
siKINOME final results.