Protein Kinase A/CREB Signaling Prevents Adriamycin-Induced Podocyte Apoptosis via Upregulation of Mitochondrial Respiratory Chain Complexes

Abstract

Previous work showed that the activation of protein kinase A (PKA) signaling promoted mitochondrial fusion and prevented podocyte apoptosis. The cAMP response element binding protein (CREB) is the main downstream transcription factor of PKA signaling. Here we show that the PKA agonist 8-(4-chlorophenylthio)adenosine 3',5'-cyclic monophosphate-cyclic AMP (pCPT-cAMP) prevented the production of adriamycin (ADR)-induced reactive oxygen species and apoptosis in podocytes, which were inhibited by CREB RNA interference (RNAi). The activation of PKA enhanced mitochondrial function and prevented the ADR-induced decrease of mitochondrial respiratory chain complex I subunits, NADH-ubiquinone oxidoreductase complex (ND) 1/3/4 genes, and protein expression. Inhibition of CREB expression alleviated pCPT-cAMP-induced ND3, but not the recovery of ND1/4 protein, in ADR-treated podocytes. In addition, CREB RNAi blocked the pCPT-cAMP-induced increase in ATP and the expression of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1-α). The chromatin immunoprecipitation assay showed enrichment of CREB on PGC1-α and ND3 promoters, suggesting that these promoters are CREB targets. , both an endogenous cAMP activator (isoproterenol) and pCPT-cAMP decreased the albumin/creatinine ratio in mice with ADR nephropathy, reduced glomerular oxidative stress, and retained Wilm's tumor suppressor gene 1 (WT-1)-positive cells in glomeruli. We conclude that the upregulation of mitochondrial respiratory chain proteins played a partial role in the protection of PKA/CREB signaling.

Interaction of CREB and PGC-1α Induces Fibronectin Type III Domain-Containing Protein 5 Expression in C2C12 Myotubes

“…We also found that overexpression of wild-type CREB, but not the CREB S133A mutant, in C2C12 myotubes upregulated Pgc-1α expression (Fig. 2e), which aligned with the previous report that CREB was a transcriptional regulator of PGC-1α expression [24].…”

Reduction of mitochondria and up regulation of pyruvate dehydrogenase kinase 4 of skeletal muscle in patients with chronic kidney disease

“…Protein samples were boiled to denature under reducing condition using NuPAGE LDS sample buffer/reducing agent (NP007/NP009, Invitrogen, New York, USA). SDS‐PAGE and immunoblot analyses were performed using standard protocols as described previously . Primary antibodies used in this study were rabbit anti‐human phospho‐PDH(S293) antibody (1:2000; Abcam, Cambridge, UK), rabbit anti‐human PDH antibody (1:2000; Abcam), rabbit anti‐human PDK4 antibody (1:1000, Abcam), rabbit anti‐human PGC‐1α antibody (1:1000, Cell Signaling Technology, Danvers, Massachusetts, USA), phospho‐Drp1(S637) antibody (1:1000, Cell Signaling Technology), phosphor‐Drp1(S616) antibody (1:1000, Cell Signaling Technology), Drp1 antibody (1:1000, Cell Signaling Technology), rabbit anti‐human Mfn‐1 antibody (1:100, Santa Cruz Biotechnology) and mouse anti‐human GAPDH antibody (1:1000, Santa Cruz Biotechnology).…”

“…SDS-PAGE and immunoblot analyses were performed using standard protocols as described previously. 10 Primary antibodies used in this study were rabbit anti-human phospho-PDH(S293) antibody…”

Phenomic screen identifies a role for the yeast lysine acetyltransferase NuA4 in the control of Bcy1 subcellular localization, glycogen biosynthesis, and mitochondrial morphology

“…The transcription factors Msn2 and Msn4 are activators of glycolysis [46], and they have also been linked to autophagy and mitochondrial respiration [70,74]. Finally, PKA activity has been closely linked to the control of mitochondrial processes through multiple targets [75][76][77][78][79][80][81]. Therefore, we assessed the structure of the mitochondria using Cit1-GFP in WT and eaf1Δ backgrounds in combination with snf1Δ, msn2msn4Δ, and bcy1Δ (hyperactive PKA) (Fig 5).…”