Reprogramming of cardiac phosphoproteome, proteome, and transcriptome confers resilience to chronic adenylyl cyclase-driven stress
- Laboratory of Cardiovascular Science, Intramural Research Program, National Institute on Aging, National Institutes of Health, United States
- Department of Immunology, St. Jude Children’s Research Hospital, United States
Figures
Figure 1 with 2 supplements
Cardiac-specific overexpression of adenylyl cyclase type 8 (TGAC8) reprograms the left ventricle (LV) phosphorylation state.
(A) Principal component analysis (PCA) plot of phosphoproteome in TGAC8 vs. WT LVs. (B) Heatmap of scaled expressions of phosphorylated proteins. (C) Volcano plot of phosphoproteome. Red dots …
Figure 1—figure supplement 1
The enriched pathway, Cardiac Hypertrophy Signaling (Enhanced), from the phosphoproteome.
Cardiac Hypertrophy Signaling (Enhanced) is visualized using ingenuity pathway analysis (IPA). Dots with purple outlines are identified phosphoproteins or protein complexes. Red and green fill …
Figure 1—figure supplement 2
The regulatory pathway is under the control of a predicted upstream kinase, PDPK1.
PDPK1 in the center is one representative of predicted upstream kinases. Surrounding dots represent identified phosphoproteins differentially regulated in phosphoproteome. Meanings of colors refer …
Figure 2
Characteristics of upregulated phosphopeptides.
(A) Composition of phosphosites that reside on the upregulated phosphopeptides. (B) Sequence logo diagrams of upregulated phosphopeptides centered on the phosphosites and flanked by six amino acids …
Figure 3
Integrative analysis of transcriptome, proteome, and phosphoproteome.
(A) Circos plot of gene expression at different levels from transcription, to protein, and to phosphorylation. Bars in each layer of circle represent changes in significantly regulated molecules …
Figure 4
Coordination of phosphoproteome and transcriptome.
(A) Gene ontology (GO) term enrichments of biological processes and molecular functions about phosphorylation and dephosphorylation from transcriptome. (B–C) Changes of kinases (B) and phosphatases …
Figure 5
Analysis of normalized phosphoproteome.
(A) Gene ontology (GO) term enrichments of biological processes and molecular functions about phosphorylation and dephosphorylation from the present proteome. (B–C) Changes of kinases (B) and …
Figure 6 with 1 supplement
Coordinated regulation of transcriptome, proteome, and phosphoproteome on PI3K/AKT and ERK/MAPK signaling pathways and protein synthesis and degradation.
(A) Canonical pathway enrichment of phosphoproteome only with cutoffp-value <0.05. The darker the color is, the greater the absolute (z-score) is, and the more significantly enriched the pathway is. …
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Figure 6—source data 1
The full raw unedited gels and blots and figures with the uncropped gels or blots with the relevant bands associated with Figure 6C and D.
- https://cdn.elifesciences.org/articles/88732/elife-88732-fig6-data1-v1.zip
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Figure 6—source data 2
The full raw unedited gels and blots and figures with the uncropped gels or blots with the relevant bands associated with Figure 6E.
- https://cdn.elifesciences.org/articles/88732/elife-88732-fig6-data2-v1.zip
Figure 6—figure supplement 1
Cross-verification of activated PI3K/AKT and ERK/MAPK signaling pathways and enhanced protein synthesis and degradation from our previous study.
(A) Western blots of one phosphorylation site on isoform 2 of AKT protein for verification of phosphorylation and activation of PI3K/AKT signaling pathway in cardiac-specific overexpression of …
Figure 7
Analyses of proteins changed discordantly in proteome and phosphoproteome.
(A) Venn diagram of significantly regulated proteins in total proteome and significantly regulated phosphoproteins in unnormalized phosphoproteome that were conducted concurrently in the same …
Figure 8
Deduced schematic of coordination of phosphoproteome with transcriptome and proteome in cardiac-specific overexpression of adenylyl cyclase type 8 (TGAC8) mouse left ventricle (LV).
Based on our results, we deduced a mechanism of reprogramed phosphoproteome in TGAC8 mouse LV: First, TGAC8 mouse induces a PKA-dependent phosphorylation via AC8-cAMP-PKA axis. Many cytoplasmic …
Figure 9
Reprogrammed concentric performance and protection circuitry in response to the constitutive challenge of marked cardiac-specific overexpression of adenylyl cyclase type 8 (TGAC8).
Based on the present phosphoproteome, we optimized our previously deduced mechanism graph and labeled findings from the phosphoproteome using red stars. TGAC8 mouse induces a chronic cardiac …
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