Figure 7: GCN5 phosphorylation at Ser275 drives its substrate switch. | Nature Communications

Figure 7: GCN5 phosphorylation at Ser275 drives its substrate switch.

From: The GCN5-CITED2-PKA signalling module controls hepatic glucose metabolism through a cAMP-induced substrate switch

Figure 7

(a) AML12 cells expressing FLAG-tagged GCN5(WT) or GCN5(S275A) were exposed to pCPT-cAMP with or without H89 for 30 min and then subjected to IP with antibodies to phosphorylated PKA substrates followed by immunoblot analysis with antibodies to FLAG. (b) Effect of the S275A mutation of FLAG-GCN5 on in vitro acetylation of histone H3 in AML12 cells treated with pCPT-cAMP (1 h). (c) Acetylation of FLAG–PGC-1α in AML12 cells expressing GCN5(WT) or GCN5(S275A) with or without HA-CITED2. (d) Effects of the S275D mutation of GCN5 on histone H3 and PGC-1α acetyltransferase activities in AML12 cells. (e) Effect of the S275D mutation of FLAG-GCN5 on interaction with V5-tagged PGC-1α in AML12 cells. (f) Effects of the S275A and S275D mutations of FLAG-GCN5 on interaction with HA-CITED2 in AML12 cells. (g) The S275A mutation of GCN5 blocks the pCPT-cAMP-induced dissociation of HA-CITED2 from FLAG-GCN5 in AML12 cells. All data are representative of at least three independent experiments. Adenoviral vectors were used for these experiments.

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